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pEN_TT 3xFLAG-MAVS-Ala148

158630

Gateway entry vector for an inducible 3xFLAG-tagged MAVS mutant

Kevin Janes

Lopacinski et al Cell Syst. 2021 Apr 21;12(4):304-323.e13. doi: 10.1016/j.cels.2021.02.004. Epub 2021 Mar 18.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/311685/ed97fa9d-d1e3-4e04-af5b-0fec9f848e1f/addgene-plasmid-158630-sequence-311685.gbk

full

LOCUS sequence-311685- 5481 bp DNA circular SYN 26-AUG-2024 DEFINITION Gateway entry vector for an inducible 3xFLAG-tagged MAVS mutant. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5481) AUTHORS Lopacinski AB, Sweatt AJ, Smolko CM, Gray-Gaillard E, Borgman CA, Shah M, Janes KA TITLE Modeling the complete kinetics of coxsackievirus B3 reveals human determinants of host-cell feedback. JOURNAL Cell Syst. 2021 Apr 21;12(4):304-323.e13. doi: 10.1016/j.cels.2021.02.004. Epub 2021 Mar 18. PUBMED 33740397 REFERENCE 2 (bases 1 to 5481) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cels.2021.02.004"; journalName: "Cell Syst"; date: "2021-04-21- 21"; volume: "12"; issue: "4"; pages: "304-323.e13" FEATURES Location/Qualifiers source 1..5481 /mol_type="other DNA" /organism="synthetic DNA construct" terminator 33..60 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind 68..91 /label=pENTR-F /note="pENTR vectors, forward primer" protein_bind 110..209 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 241..555 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 249..267 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 285..303 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 320..338 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 356..374 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 392..410 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 427..445 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 463..481 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 528..552 /label=LNCX /note="Human CMV promoter, forward primer" CDS 564..629 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" protein_bind complement(2294..2393) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2505..2524) /label=pENTR-R /note="pENTR vectors, reverse primer" primer_bind complement(2583..2602) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" CDS complement(3176..3709) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(3898..3926) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" rep_origin 4583..5171 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5072..5091 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 5336..5422 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg atggcctttt 61 tgcgtttcta caaactcttc ctgttagtta gttacttaag ctcgggcccc aaataatgat 121 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gcttttttat 181 aatgccaact ttgtacaaaa aagcaggctt taaaggaacc aattcagtcg actggatccc 241 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag 301 agaacgatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct 361 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg 421 agtttatccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag 481 aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 541 aaccgtcaga tcgccactag atggactaca aagaccatga cggtgattat aaagatcatg 601 acatcgatta caaggatgac gatgacaaga ctagtatgcc gtttgctgaa gacaagacct 661 ataagtatat ctgccgcaat ttcagcaatt tttgcaatgt ggatgttgta gagattctgc 721 cttacctgcc ctgcctcaca gcaagagacc aggatcgact gcgggccacc tgcacactct 781 cagggaaccg ggacaccctc tggcatctct tcaataccct tcagcggcgg cccggctggg 841 tggagtactt cattgcggca ctgaggggct gtgagctagt tgatctcgcg gacgaagtgg 901 cctctgtcta cgagagctac cagcctcgga cctcggaccg tcccccagac ccactggagc 961 caccgtcact tcctgctgag aggccagggc cccccacacc tgctgcggcc cacagcatcc 1021 cctacaacag ctgcagagag aaggagccaa gttaccccat gcctgtccag gagaccgcgg 1081 cgccagagtc cccaggagag aattcagagc aagccctgca gacgctcagc cccagagcca 1141 tcccaaggaa tccagatggt ggccccctgg agtcctcctc tgacctggca gccctcagcc 1201 ctctgacctc cagcgggcat caggagaagg acacagaact gggcagtacc cacacagcag 1261 gtgcgacctc cagcctcaca ccatcccgtg ggcctgtgtc tccatctgtc tccttccagc 1321 ccctggcccg ttccaccccc agggcaagcc gcttgcctgg acccacaggg tcagttgtat 1381 ctactggcac ctccttctcc tcctcatccc ctggcttggc ctctgcaggg gctgcagagg 1441 gtaaacaggg tgcagagagt gaccaggccg agcctatcat ctgctccagt ggggcagagg 1501 cacctgccaa ctctctgccc tccaaagtgc ctaccacctt gatgcctgtg aacacagtgg 1561 ccctgaaagt gcctgccaac ccagcatctg tcagcacagt gccctccaag ttgccaacta 1621 gctcaaagcc ccctggtgca gtgccttcta atgcgctcac caatccagca ccatccaaat 1681 tgcccatcaa ctcaacccgt gctggcatgg tgccatccaa agtgcctact agcatggtgc 1741 tcaccaaggt gtctgccagc acagtcccca ctgacgggag cagcagaaat gaggagaccc 1801 cagcagctcc aacacccgcc ggcgccactg gaggcagctc agcctggcta gacagcagct 1861 ttgagaatag gggccttggg tcggagctga gtaagcctgg cgtgctggca tcccaggtag 1921 acagcccgtt ctcgggctgc ttcgaggatc ttgccatcag tgccagcacc tccttgggca 1981 tggggccctg ccatggccca gaggagaatg agtataagtc cgagggcacc tttgggatcc 2041 acgtggctga gaaccccagc atccagctcc tggagggcaa ccctgggcca cctgcggacc 2101 cggatggcgg ccccaggcca caagccgacc ggaagttcca ggagagggag gtgccatgcc 2161 acaggccctc acctggggct ctgtggctcc aggtggctgt gacaggggtg ctggtagtca 2221 cactcctggt ggtgctgtac cggcggcgtc tgcactagca attggaagac taatgcgttt 2281 tcgagatatc tagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta 2341 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca 2401 gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat 2461 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag 2521 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata 2581 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta 2641 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga 2701 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat 2761 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa 2821 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct 2881 ggcagtgtcc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga 2941 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag 3001 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa 3061 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct 3121 tatttttgac gaggggaaat taggccggga agccgatctc ggcttgaacg aattgttagg 3181 tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat 3241 agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa 3301 gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt 3361 gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg 3421 gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga 3481 tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt 3541 aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga 3601 cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg 3661 ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc 3721 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata 3781 gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg 3841 ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt 3901 ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg 3961 cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac 4021 gagcgcaagg tttcggtctc cacgcatcgt caggcattgg cggccttgct gttcttctac 4081 ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg 4141 cggcgcttgc cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa 4201 ggcgagcatc gtttgttcgc ccaggactct agctatagtt ctagtggttg gctactaata 4261 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta 4321 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt 4381 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa 4441 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc 4501 gcaagctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 4561 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 4621 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 4681 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 4741 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 4801 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 4861 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 4921 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 4981 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 5041 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 5101 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 5161 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 5221 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcta 5281 gcatggatct cggggacgtc taactactaa gcgagagtag ggaactgcca ggcatcaaat 5341 aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa 5401 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc 5461 cggagggtgg cgggcaggac g //

pDEST40-2XHA-Src-K298M-Y419F-Y530F

140320

Expresses HA-tagged human Src-K298M-Y419F-Y530F in mammalian cells for structure-function studies

Mark Moasser

Spassov et al Cell Rep. 2018 Oct 9;25(2):449-463.e4. doi: 10.1016/j.celrep.2018.09.035.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/291904/76fbec73-1ec6-4346-b149-778bd20aba4b/addgene-plasmid-140320-sequence-291904.gbk

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LOCUS sequence-291904- 7181 bp DNA circular SYN 26-AUG-2024 DEFINITION Expresses HA-tagged human Src-K298M-Y419F-Y530F in mammalian cells for structure-function studies. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7181) AUTHORS Spassov DS, Ruiz-Saenz A, Piple A, Moasser MM TITLE A Dimerization Function in the Intrinsically Disordered N-Terminal Region of Src. JOURNAL Cell Rep. 2018 Oct 9;25(2):449-463.e4. doi: 10.1016/j.celrep.2018.09.035. PUBMED 30304684 REFERENCE 2 (bases 1 to 7181) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.celrep.2018.09"; journalName: "Cell Rep"; date: "2018-10-9- 9"; volume: "25"; issue: "2"; pages: "449-463.e4" FEATURES Location/Qualifiers source 1..7181 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 911..935 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2562..2586) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2597..2623 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2627..2653 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2679..2720 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2730..2747 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2770..2787) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2776..3000 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3046..3474 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3133..3152) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3343..3364 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3483..3503) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3488..3817 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3668..3803 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3730..3749 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3884..4678 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3938..3957) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4548..4567 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4854..4975 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4891..4910) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4945..4964 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5024..5040) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5024..5040) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5037..5059) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5048..5064 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5072..5102) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5117..5138 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5255..5272) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5426..6014) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5506..5525) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6185..7045) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6808..6827 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7046..7150) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagctatca acaagtttgt acaaaaaagc aggcttcgaa ggagatagaa ccatgggtag 961 caacaagagc aagcccaagg atgccagcca gcggcgccgc agcctggagc ccgccgagaa 1021 cgtgcacggc gctggcgggg gcgctttccc cgcctcgcag acccccagca agccagcctc 1081 ggccgacggc caccgcggcc ccagcgcggc cttcgccccc gcggccgccg agcccaagct 1141 gttcggaggc ttcaactcct cggacaccgt cacctccccg cagagggcgg gcccgctggc 1201 cggtggagtg accacctttg tggccctcta tgactatgag tctaggacgg agacagacct 1261 gtccttcaag aaaggcgagc ggctccagat tgtcaacaac acagagggag actggtggct 1321 ggcccactcg ctcagcacag gacagacagg ctacatcccc agcaactacg tggcgccctc 1381 cgactccatc caggctgagg agtggtattt tggcaagatc accagacggg agtcagagcg 1441 gttactgctc aatgcagaga acccgagagg gaccttcctc gtgcgagaaa gtgagaccac 1501 gaaaggtgcc tactgcctct cagtgtctga cttcgacaac gccaagggcc tcaacgtgaa 1561 gcactacaag atccgcaagc tggacagcgg cggcttctac atcacctccc gcacccagtt 1621 caacagcctg cagcagctgg tggcctacta ctccaaacac gccgatggcc tgtgccaccg 1681 cctcaccacc gtgtgcccca cgtccaagcc gcagactcag ggcctggcca aggatgcctg 1741 ggagatccct cgggagtcgc tgcggctgga ggtcaagctg ggccagggct gctttggcga 1801 ggtgtggatg gggacctgga acggtaccac cagggtggcc atcatgaccc tgaagcctgg 1861 cacgatgtct ccagaggcct tcctgcagga ggcccaggtc atgaagaagc tgaggcatga 1921 gaagctggtg cagttgtatg ctgtggtttc agaggagccc atttacatcg tcacggagta 1981 catgagcaag gggagtttgc tggactttct caagggggag acaggcaagt acctgcggct 2041 gcctcagctg gtggacatgg ctgctcagat cgcctcaggc atggcgtacg tggagcggat 2101 gaactacgtc caccgggacc ttcgtgcagc caacatcctg gtgggagaga acctggtgtg 2161 caaagtggcc gactttgggc tggctcggct cattgaagac aatgagttca cggcgcggca 2221 aggtgccaaa ttccccatca agtggacggc tccagaagct gccctctatg gccgcttcac 2281 catcaagtcg gacgtgtggt ccttcgggat cctgctgact gagctcacca caaagggacg 2341 ggtgccctac cctgggatgg tgaaccgcga ggtgctggac caggtggagc ggggctaccg 2401 gatgccctgc ccgccggagt gtcccgagtc cctgcacgac ctcatgtgcc agtgctggcg 2461 gaaggagcct gaggagcggc ccaccttcga gtacctgcag gccttcctgg aggactactt 2521 cacgtccacc gagccccagt tccagcccgg ggagaacctc gacccagctt tcttgtacaa 2581 agtggttgat ctagagtacc catacgatgt tccagattac gctggatacc catacgatgt 2641 tccagattac gcttgatcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc 2701 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc 2761 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg 2821 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa 2881 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca 2941 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg 3001 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg 3061 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 3121 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 3181 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 3241 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 3301 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 3361 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga 3421 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct 3481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 3601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 3661 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 3721 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta 3781 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc 3841 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 3901 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 3961 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 4021 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 4081 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 4141 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 4201 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 4261 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 4321 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 4381 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 4441 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 4501 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 4561 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 4621 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc 4681 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 4741 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 4801 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 4861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 4921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 4981 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 5041 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 5101 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 5161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 5221 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 5281 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 5341 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 5401 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 5461 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 5521 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 5581 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 5641 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 5701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 5761 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 5821 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 5881 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 5941 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 6001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 6061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 6121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 6181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 7081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 7141 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //

pNP1 OR input B

107366

Input B for pNP1 OR gate sfGFP

James Collins

Huang et al Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. eCollection 2018 Aug.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/209408/6e44caf1-b32e-4796-86a0-be6347141558/addgene-plasmid-107366-sequence-209408.gbk

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LOCUS sequence-209408- 2116 bp DNA circular SYN 26-AUG-2024 DEFINITION Input B for pNP1 OR gate sfGFP. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2116) AUTHORS Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ TITLE BioBits Explorer: A modular synthetic biology education kit. JOURNAL Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. eCollection 2018 Aug. PUBMED 30083608 REFERENCE 2 (bases 1 to 2116) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; date: "2018-08-1"; volume: "4(8):eaat5105. doi"; pages: " 10.1126/sciadv.aat5105. eCollection 2018 Aug" FEATURES Location/Qualifiers source 1..2116 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 26..45 /label=T7 /note="T7 promoter, forward primer" promoter 26..44 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" terminator 108..155 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin complement(373..961) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(453..472) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1133..1993) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1756..1775 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1994..2098) /gene="bla" /label=AmpR promoter ORIGIN 1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggagtcg gcgtggactc 61 gcacgccgac taaacataac cataaagcgc gatcttaagt tcatacccta gcataacccc 121 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggaa 181 gcttggcact ggccgaccgg ggtcgagcac tgactcgctg cgctcggtcg ttcggctgcg 241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 661 cttatccggt aactatcgtc ttgagtccaa cccgctaaga cacgacttat cgccactggc 721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 781 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacagat 1021 ccgggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 1081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 1141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 1201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 1261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 1321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 1381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 1441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 1501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 1561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 1621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 1681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 1741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 1801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 1861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 1921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 1981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 2041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 2101 catttccccg aaaagt //

FSW-HRP-V5-MDGA1

82541

Lentiviral vector for expressing HRP-MDGA1 in neurons

Alice Ting

Loh et al Cell. 2016 Aug 25;166(5):1295-1307.e21. doi: 10.1016/j.cell.2016.07.041.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/163055/52e361c1-43aa-4c92-9d35-46ebd4db58cf/addgene-plasmid-82541-sequence-163055.gbk

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LOCUS sequence-163055- 12366 bp DNA circular SYN 26-AUG-2024 DEFINITION Lentiviral vector for expressing HRP-MDGA1 in neurons. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12366) AUTHORS Loh KH, Stawski PS, Draycott AS, Udeshi ND, Lehrman EK, Wilton DK, Svinkina T, Deerinck TJ, Ellisman MH, Stevens B, Carr SA, Ting AY TITLE Proteomic Analysis of Unbounded Cellular Compartments: Synaptic Clefts. JOURNAL Cell. 2016 Aug 25;166(5):1295-1307.e21. doi: 10.1016/j.cell.2016.07.041. PUBMED 27565350 REFERENCE 2 (bases 1 to 12366) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cell.2016.07"; journalName: "Cell"; date: "2016-08-25- 25"; volume: "166"; issue: "5"; pages: "1295-1307.e21" FEATURES Location/Qualifiers source 1..12366 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2098..2142 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2636..3083 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 3091..3100 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 4144..4185 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 7058..7646 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7111..7131) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7529..7540) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(7649..7665) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7650..7666) /label=pBluescriptKS /note="For pBluescript vector" LTR 8171..8351 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(8377..8394) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8383..8607 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 8653..9081 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8740..8759) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8950..8971 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(9090..9110) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9095..9424 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9275..9410 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9337..9356 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 9472..9519 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 9538..9912 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 10042..10163 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10079..10098) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10133..10152 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10212..10228) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10212..10228) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10225..10247) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10236..10252 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(10260..10290) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10305..10326 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(10443..10460) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10614..11202) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10694..10713) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11373..12233) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 11996..12015 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12234..12338) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagcacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttaatta ataccgccat gcatctgcag agggccctgc gtatgagtgc 2641 aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg 2701 acccactgga caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg 2761 ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag 2821 tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga 2881 cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg 2941 ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc 3001 atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag 3061 tcgtgtcgtg cctgagagcg cagtctagac gccaccatgg atttcctgct gctcggtctc 3121 tgtctatact ggctgctgag gaggccctcg ggggtggtct tgtgtctgct gggggcctgc 3181 tttcagatgc tgcccgccgc tagccagctt actccaactt tctacgataa ctcttgtcct 3241 aacgtgagca acatcgtgcg agatactatc gtgaacgagt tgcggtctga tccgaggata 3301 gccgcttcaa ttttgcggct gcatttccac gattgttttg taaacggctg tgatgcttcc 3361 atcctcctcg acaataccac ctctttcagg actgaaaagg atgccttcgg taacgcgaat 3421 agtgctagag ggtttcccgt gatcgataga atgaaagccg ctgtggagag cgcctgcccc 3481 cggacagtta gttgtgcaga ccttctgacc atagcggccc agcaatccgt gacgctggcc 3541 ggtggcccca gctggagggt ccccctggga aggagggact ctctgcaggc ttttctggac 3601 ttggcaaacg caaacctgcc tgctcccttc ttcacccttc cccagctgaa ggattccttc 3661 aggaatgtcg gcctgaaccg aagtagtgat ttggtggctc tgtccggagg ccacacattc 3721 ggcaagaatc agtgcaggtt catcatggac agactgtata acttttccaa caccggcttg 3781 cctgacccta ccctgaacac aacatacttg cagactctca gagggctgtg tccgctcaat 3841 ggcaacttgt ctgcactggt tgactttgac ctgcgcaccc caaccatctt cgacaataag 3901 tattacgtca atctggagga acagaagggc ctgattcagt ccgaccagga gctgttcagt 3961 tccccaaatg ctaccgacac tattcctctt gtgcgcagct tcgctaatag cacacaaacc 4021 tttttcaatg ctttcgtgga agctatggat cggatgggaa atataacgcc cctgacgggt 4081 acgcagggac agataagact gaactgtagg gtcgtgaaca gcaactctgg ctcgggctcg 4141 ggcggtaagc ccatccccaa ccccctgctg ggcctggaca gcaccaccgg tcactgccgg 4201 ggacaaggag tctacgctcc agcccaggcg cagatcgtgc atgcgggcca ggcatgtgtg 4261 gtgaaagagg acaatatcag cgagcgtgtc tacaccatcc gggaggggga caccctcatg 4321 ctgcagtgcc ttgtaacagg gcaccctcga ccccaggtac ggtggaccaa gacggcaggt 4381 agcgcctcgg acaagttcca ggagacatcg gtgttcaacg agacgctgcg catcgagcgt 4441 attgcacgca cgcagggcgg ccgctactac tgcaaggctg agaacggcgt gggggtgccg 4501 gccatcaagt ccatccgcgt ggacgtgcag tacctggatg agccaatgct gacggtgcac 4561 cagacggtga gcgatgtgcg aggcaacttc taccaggaga agacggtgtt cctgcgctgt 4621 actgtcaact ccaacccgcc tgcccgcttc atctggaagc ggggttccga taccctatcc 4681 cacagccagg acaatggggt tgacatctat gagcccctct acactcaggg ggagaccaag 4741 gtcctgaagc tgaagaacct gcggccccag gactatgcca gctacacctg ccaggtgtct 4801 gtgcgtaacg tgtgcggcat cccagacaag gccatcacct tccggctcac caacaccacg 4861 gcaccaccag ccctgaagct gtctgtgaac gaaactctgg tggtgaaccc tggggagaat 4921 gtgacggtgc agtgtctgct gacaggcggt gatcccctcc cccagctgca gtggtcccat 4981 gggcctggcc cactgcccct gggtgctctg gcccagggtg gcaccctcag catcccttca 5041 gtgcaggccc gggactctgg ctactacaac tgcacagcca ccaacaatgt gggcaaccct 5101 gccaagaaga ctgtcaacct gctggtgcga tccatgaaga acgctacatt ccagatcact 5161 cctgacgtga tcaaagagag tgagaacatc cagctgggcc aggacctgaa gctatcgtgc 5221 cacgtggatg cagtgcccca ggagaaggtg acctaccagt ggttcaagaa tggcaagccg 5281 gcacgcatgt ccaagcggct gctggtgacc cgcaatgatc ctgagctgcc cgcagtcacc 5341 agcagcctag agctcattga cctgcacttc agtgactatg gcacctacct gtgcatggct 5401 tctttcccag gggcacccgt gcccgacctc agcgtcgagg tcaacatctc ctctgagaca 5461 gtgccgccca ccatcagtgt gcccaagggt agggccgtgg tgaccgtgcg cgagggatcg 5521 cctgccgagc tgcaatgcga ggtgcggggc aagccgcggc cgccagtgct ctggtcccgc 5581 gtggacaagg aggctgcact gctgccctcg gggctgcccc tggaggagac tccggacggg 5641 aagctgcggc tggagcgagt gagccgagac atgagcggga cctaccgctg ccagacggcc 5701 cgctataatg gcttcaacgt gcgcccccgt gaggcccagg tgcagctgaa cgtgcagttc 5761 ccgccggagg tggagcccag ttcccaggac gtgcgccagg cgctgggccg gcccgtgctc 5821 ctgcgctgct cgctgctgcg aggcagcccc cagcgcatcg cctcggctgt gtggcgtttc 5881 aaagggcagc tgctgccgcc gccgcctgtt gttcccgccg ccgccgaggc gccggatcac 5941 gcggagctgc gcctcgacgc cgtaactcgc gacagcagcg gcagctacga gtgcagcgtc 6001 tccaacgatg tgggctcggc tgcctgcctc ttccaggtct ccgccaaagc ctacagcccg 6061 gagttttact tcgacacccc caaccccacc cgcagccaca agctgtccaa gaactactcc 6121 tacgtgctgc agtggactca gagggagccc gacgctgtcg accctgtgct caactacaga 6181 ctcagcatcc gccagttgaa ccagcacaat gcggtggtca aggccatccc ggtccggcgt 6241 gtggagaagg ggcagctgct ggagtacatc ctgaccgatc tccgtgtgcc ccacagctat 6301 gaggtccgcc tcacacccta taccaccttc ggggctggtg acatggcctc ccgcatcatc 6361 cactacacag agcccatcaa ctctccgaac ctttcagaca acacctgcca ctttgaggat 6421 gagaagatct gtggctatac ccaggacctg acagacaact ttgactggac gcggcagaat 6481 gccctcaccc agaaccccaa acgctccccc aacactggtc cccccaccga cataagtggc 6541 acccctgagg gctactacat gttcatcgag acatcgaggc ctcgggagct gggggaccgt 6601 gcaaggttag tgagtcccct ctacaatgcc agcgccaagt tctactgtgt ctccttcttc 6661 taccacatgt acgggaaaca catcggctcc ctcaacctcc tggtgcggtc ccggaacaaa 6721 ggggctctgg acacgcacgc ctggtctctc agtggcaata agggcaatgt gtggcagcag 6781 gcccatgtgc ccatcagccc cagtgggccc ttccagatta tttttgaggg ggttcgaggc 6841 ccgggctacc tgggggatat tgccatagat gacgtcacac tgaagaaggg ggagtgtccc 6901 cggaagcaga cggatcccaa taaagtggtg gtgatgccgg gcagtggagc cccctgccag 6961 tccagcccac agctgtgggg gcccatggcc atcttcctct tggcgttgca gagatgaggc 7021 gcgcccctgc agggaattcg atatcaagct tatcgataat caacctctgg attacaaaat 7081 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 7141 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 7201 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 7261 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 7321 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 7381 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 7441 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 7501 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 7561 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 7621 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca 7681 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc 7741 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac 7801 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 7861 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta 7921 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg 7981 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga 8041 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt 8101 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc 8161 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 8221 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 8281 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 8341 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc 8401 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca 8461 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta 8521 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc 8581 atgctgggga tgcggcgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta 8641 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 8701 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 8761 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 8821 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 8881 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 8941 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 9001 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 9061 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc 9121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 9181 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 9241 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 9301 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 9361 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 9421 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat 9481 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg 9541 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag 9601 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg 9661 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac 9721 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc 9781 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg 9841 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag 9901 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg 9961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 10021 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 10081 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 10141 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 10201 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 10261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 10321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 10381 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 10441 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 10501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 10561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 10621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 10681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 10741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 10801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 10861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 10921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 10981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 11041 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 11101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 11161 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 11221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 11281 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 11341 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 11401 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 11461 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 11521 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 11581 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 11641 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 11701 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 11761 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 11821 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 11881 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 11941 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 12001 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 12061 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 12121 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 12181 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 12241 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 12301 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 12361 cctgac //

pDONR221_SLC30A2

132113

Gateway entry clone with codon-optimized ORF sequence for subcloning into custom expression vectors

Superti-Furga et al Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 10.1038/d41573-020-00056-6.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/265280/3a99d68f-ee01-4d58-8d83-81b9a2028c83/addgene-plasmid-132113-sequence-265280.gbk

full

LOCUS sequence-265280- 3519 bp DNA circular SYN 26-AUG-2024 DEFINITION Gateway entry clone with codon-optimized ORF sequence for subcloning into custom expression vectors. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3519) AUTHORS Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov V, Wolf G, Crawford M, Everley R, ... Steppan CM TITLE The RESOLUTE consortium: unlocking SLC transporters for drug discovery. JOURNAL Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 10.1038/d41573-020-00056-6. PUBMED 32265506 REFERENCE 2 (bases 1 to 3519) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; pages: "429-430" FEATURES Location/Qualifiers source 1..3519 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator complement(268..295) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(387..473) /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1644..1743) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1760..1779) /label=T7 /note="T7 promoter, forward primer" promoter complement(1761..1779) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1784..1800) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1784..1800) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1902..1921) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1913..2722 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1980..1999) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2869..3457 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3358..3377 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agcaggctcc accatggagg ccaaggagaa gcagcacctg ctggacgcaa ggccagcaat 721 cagatcctac accggatctc tgtggcagga gggagcagga tggatcccac tgcctaggcc 781 aggactggat ctgcaggcaa tcgagctggc agcacagtcc aaccaccact gccacgcaca 841 gaagggacca gactctcact gtgatcctaa gaagggcaag gcccagagac agctgtatgt 901 ggccagcgcc atctgcctgc tgtttatgat aggcgaagta gttgagatcc tgggcgccct 961 ggtgtctgtg ctgagcatct gggtggtgac cggcgtgctg gtgtacctgg ccgtggagag 1021 gctgatctct ggcgactatg agatcgatgg cggcaccatg ctgatcacaa gcggctgtgc 1081 cgtggccgtg aatatcatca tgggactgac actgcaccag agcggacacg gacactccca 1141 cggcaccaca aaccagcagg aggagaatcc ttccgtgcgc gccgccttca tccacgtgat 1201 cggcgacttt atgcagtcta tgggcgtgct ggtggccgcc tacatcctgt atttcaagcc 1261 agagtacaag tatgtggacc ccatctgcac cttcgtgttt agcatcctgg tgctgggcac 1321 cacactgaca atcctgaggg atgtgatcct ggtgctgatg gagggcaccc ctaagggcgt 1381 ggactttaca gccgtgcgcg atctgctgct gtctgtggag ggagtggagg ccctgcacag 1441 cctgcacatc tgggcactga ccgtggcaca gccagtgctg tccgtgcaca tcgccatcgc 1501 ccagaacacc gatgcccagg ccgtgctgaa gacagccagc tcccggctgc agggcaagtt 1561 ccactttcac accgtgacaa tccagatcga ggactacagc gaggacatga aggattgcca 1621 ggcctgtcag ggcccctccg atgacccagc tttcttgtac aaagttggca ttataagaaa 1681 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat 1741 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg 1801 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc 1861 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca 1921 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg 1981 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg 2041 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt 2101 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa 2161 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac 2221 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc 2281 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg 2341 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga 2401 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact 2461 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat 2521 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg 2581 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa 2641 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt 2701 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat 2761 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc 2821 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 2881 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 2941 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 3001 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 3061 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 3121 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 3181 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 3241 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 3301 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 3361 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 3421 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 3481 acggttcctg gccttttgct ggccttttgc tcacatgtt //

pcDNA-Prom1 SV8 (-Ex19a)

184012

Mammalian Flag tagged expression clone for the mouse Prom1 splicing variant SV8 (GenBank accession BC028286). Deletion of exon 19a (amino acids 696-701).

Peter Stoilov

Matalkah et al PLoS One. 2022 Oct 10;17(10):e0274958. doi: 10.1371/journal.pone.0274958. eCollection 2022.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/362442/7f6a9064-1839-41ee-9789-a7e4a02b81d6/addgene-plasmid-184012-sequence-362442.gbk

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LOCUS sequence-362442- 7863 bp DNA circular SYN 27-AUG-2024 DEFINITION Mammalian Flag tagged expression clone for the mouse Prom1 splicing variant SV8 (GenBank accession BC028286). Deletion of exon 19a (amino acids 696-701).. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7863) AUTHORS Matalkah F, Rhodes S, Ramamurthy V, Stoilov P TITLE The mAB 13A4 monoclonal antibody to the mouse PROM1 protein recognizes a structural epitope. JOURNAL PLoS One. 2022 Oct 10;17(10):e0274958. doi: 10.1371/journal.pone.0274958. eCollection 2022. PUBMED 36215230 REFERENCE 2 (bases 1 to 7863) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One."; date: "2022-10-10"; volume: "17(10):e0274958. doi"; pages: " 10.1371/journal.pone.0274958. eCollection 2022" FEATURES Location/Qualifiers source 1..7863 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 894..903 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 3408..3431 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 3460..3684 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3730..4158 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3817..3836) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4027..4048 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4167..4187) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4172..4501 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4352..4487 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4414..4433 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4568..5362 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4622..4641) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5232..5251 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5536..5657 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5573..5592) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5627..5646 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5706..5722) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5706..5722) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5719..5741) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5730..5746 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5754..5784) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5799..5820 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5937..5954) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6108..6696) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6188..6207) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6867..7727) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7490..7509 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7728..7832) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctgccacca 901 tggctctcgt cttcagtgcc ctgctgttac tggggctgtg tggaaagatc tcttcagaag 961 gtcagcctgc attccataac actcctgggg ctatgaatta tgaattgcct accaccaaat 1021 atgagaccca agataccttc aatgctggga ttgttggccc tctctacaaa atggtgcaca 1081 tcttcctcaa cgtggtccag ccgaatgact tccctctaga tttgatcaaa aaactcatac 1141 agaacaagaa ctttgacatc tcagttgatt ccaaggagcc agaaatcata gtcttggctc 1201 tgaagattgc cctctatgag atcggagtcc ttatctgcgc catcctggga ctgctgttca 1261 ttatcctcat gcctctggtg ggctgcttct tttgtatgtg ccgttgctgc aacaaatgcg 1321 gcggagagat gcaccagcgg cagaagcaga atgcgccatg caggaggaag tgcttgggcc 1381 tctccctcct ggtgatttgt ctgctcatga gccttggcat tatatatggc tttgtggcta 1441 accagcagac caggactcgg atcaaaggga cccagaaact ggcaaagagc aatttcagag 1501 actttcaaac actcctgact gaaacaccaa agcaaattga ctatgtagtg gagcagtaca 1561 ccaacaccaa gaacaaggca ttctcagacc tggatggcat cggctccgtg ctgggaggca 1621 gaataaagga ccaactaaaa cccaaagtaa ctcctgtcct cgaagagatt aaggccatgg 1681 cgacagccat caaacagacc aaggatgccc tgcagaacat gagcagcagc ctgaaaagtc 1741 tccaagatgc agccacccag ctcaatacca acctgagctc tgtgagaaac agcatcgaga 1801 attcgctcag cagcagtgac tgtacctcag atccagccag caagatctgc gatagcatca 1861 gaccaagcct aagcagtctg gggagcagcc tcaattcaag tcagctccca tcagtggata 1921 gagaactcaa cactgttact gaagtcgaca aaactgatct ggagagcctc gtcaaaaggg 1981 ggtatacgac aattgatgaa atacccaata caatacaaaa ccaaactgtg gatgtcatca 2041 aagacgtcaa aaataccttg gactccatta gctccaacat taaggacatg agccaaagta 2101 ttcctattga ggatatgctg ttacaggtct cccattacct taataacagc aacagatact 2161 taaaccagga gctgcccaag ctggaagaat atgactcgta ctggtggctg ggtggcttga 2221 ttgtctgctt tctgctgact ctcattgtga ccttcttttt cctgggcttg ctgtgtggtg 2281 tgtttggcta tgacaagcat gccaccccaa ctagaagagg ctgtgtgtcc aacactggag 2341 gcatcttcct catggctggg gttggattcg gcttcctttt ttgctggata ttgatgatcc 2401 ttgtggttct tacgtttgtt gttggtgcaa atgtggaaaa gttgctctgc gaaccttatg 2461 aaaacaagaa attattacag gttttggaca ctccctatct gctcaaggaa caatggcaat 2521 tttatctttc tggcatgcta ttcaataacc cagacattaa catgaccttt gagcaagtct 2581 acagggattg caaaagaggt cgaggtatat atgctgcttt tcagcttgag aatgtcgtca 2641 acgtcagtga tcatttcaac attgaccaga tttctgaaaa cataaatacg gagttggaaa 2701 acctgaatgt gaacattgat agcattgaac tgttggataa cacaggaagg aagagcctcg 2761 aggactttgc acattctggg atagatacaa tcgattattc cacatacttg aaggagactg 2821 agaaatcccc tactgaagtg aatctgctga catttgcctc taccctggaa gcaaaagcaa 2881 accagttgcc tgaaggaaag ctgaaacagg ccttcttact ggatgtacag aatataagag 2941 ccatccacca gcatctcctc cctcctgtgc agcaatcact gaatacgtta agacaaagtg 3001 tctggaccct ccagcaaaca agcaacaagt tgccggagaa agtgaagaag atccttgcct 3061 ctttggactc tgttcagcat ttcctcacca ataacgtttc cctcatcgtt atcggggaaa 3121 cgaagaagtt tgggaaaaca atactaggct actttgaaca ttatctgcac tgggtctttt 3181 atgccatcac agagaagatg acatcctgca aacccatggc caccgcgatg gactctgctg 3241 ttaatggcat tctgtgtggc tatgttgcgg accctctgaa tttgttctgg ttcggcatag 3301 ggaaagccac ggtgctctta cttccggctg taatcattgc tatcaagctg gccaagtact 3361 atcgcaggat ggattcagag gatgtatacg acgacccgtc tcgatacgac tacaaagacg 3421 atgacgacaa gtgagtttaa acccgctgat cagcatcgac tgtgccttct agttgccagc 3481 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg 3541 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc 3601 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg 3661 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg 3721 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 3781 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 3841 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 3901 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 3961 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 4021 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 4081 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 4141 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 4261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 4321 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 4381 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 4441 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 4501 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc 4561 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 4621 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 4681 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 4741 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 4801 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 4861 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 4921 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 4981 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 5041 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 5101 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 5161 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 5221 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 5281 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 5341 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg 5401 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 5461 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 5521 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 5581 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 5641 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 5701 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 5761 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 5821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 5881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 5941 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 6001 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 6061 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 6121 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 6181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 6241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 6301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 6361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 6421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 6481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 6541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 6601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 6661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 6721 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 6781 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 6841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 6901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 6961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 7021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 7081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 7141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 7201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 7261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 7321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 7381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 7441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 7501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 7561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 7621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 7681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 7741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 7801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 7861 gtc //

pCRUSH

54480

(Empty Backbone)Mouse genomic ROSA26 targeting vector, expresses EGFP, cre/lox conditional U6 promoter pgk-puro

Laurie Jackson-Grusby

Brown et al Genesis. 2014 Jan;52(1):39-48. doi: 10.1002/dvg.22718. Epub 2013 Oct 26.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/91228/0adc9959-6673-4ec3-8c94-ec281530630c/addgene-plasmid-54480-sequence-91228.gbk

partial

LOCUS sequence-91228-e 836 bp DNA linear UNA 26-AUG-2024 DEFINITION Mouse genomic ROSA26 targeting vector, expresses EGFP, cre/lox conditional U6 promoter pgk-puro. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 836) AUTHORS Brown JR, Zetsche B, Jackson-Grusby L TITLE RUSH and CRUSH: a rapid and conditional RNA interference method in mice. JOURNAL Genesis. 2014 Jan;52(1):39-48. doi: 10.1002/dvg.22718. Epub 2013 Oct 26. PUBMED 24166816 REFERENCE 2 (bases 1 to 836) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1002/dvg.22718"; journalName: "Genesis"; date: "2014-01"; volume: "52"; issue: "1"; pages: "39-48" FEATURES Location/Qualifiers source 1..836 /mol_type="genomic DNA" /organism="unspecified" misc_feature 204..307 /label=Ad SA /note="adenovirus major late transcript splice acceptor (Friedrich and Soriano, 1991)" regulatory 355..364 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 361..825 /codon_start=1 /product="improved N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VN155(I152L) /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA" primer_bind complement(406..427) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(667..686) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" ORIGIN 1 taggcgggga gaaggccgca cccttctccg gaggggggag gggagtgttg caataccttt 61 ctgggagttc tctgctgcct cctggcttct gaggaccgcc ctgggcctgg gagaatccct 121 tccccctctt ccctcgtgat ctgcaactcc agtctttcta cggtcgaggc tagaactagt 181 ggatcccccg ggctgcagat ctgtagggcg cagtagtcca gggtttcctt gatgatgtca 241 tacttatcct gtcccttttt tttccacagc tcgcggttga ggacaaactc ttcgcggtct 301 ttccagtggg gatcgacggt atcgatggat ccgaattcag tggatccacc ggtcgccacc 361 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 421 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 481 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 541 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 601 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 661 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 721 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 781 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaa //

pNC-RFP-C

193413

(Empty Backbone)Nimble cloning-compatible vector for protein subcellular localization in plants

Pu Yan

Yan et al Plant Commun. 2023 Mar 13;4(2):100471. doi: 10.1016/j.xplc.2022.100471. Epub 2022 Nov 9.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/380181/b2050e65-3040-40cc-b3ad-12d2f380fe07/addgene-plasmid-193413-sequence-380181.gbk

full

LOCUS sequence-380181- 5454 bp DNA circular SYN 27-AUG-2024 DEFINITION Nimble cloning-compatible vector for protein subcellular localization in plants. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5454) AUTHORS Yan P, Tuo D, Shen W, Deng H, Zhou P, Gao X TITLE A Nimble Cloning-compatible vector system for high-throughput gene functional analysis in plants. JOURNAL Plant Commun. 2023 Mar 13;4(2):100471. doi: 10.1016/j.xplc.2022.100471. Epub 2022 Nov 9. PUBMED 36352791 REFERENCE 2 (bases 1 to 5454) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Commun."; date: "2023-03-13"; volume: "4(2):100471. doi"; pages: " 10.1016/j.xplc.2022.100471. Epub 2022 Nov 9" FEATURES Location/Qualifiers source 1..5454 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 184..529 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind 472..492 /label=35S promoter /note="CaMV 35S promoter, forward primer" promoter 725..827 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 858..1163 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 1042..1061 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 1221..1898 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" CDS 1221..1898 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" primer_bind complement(1351..1369) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1619..1638 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1813..1836 /label=DsRed1-C /note="DsRed1, forward primer" terminator 1921..2173 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(2191..2207) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2191..2207) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2244..2264) /label=T3 /note="T3 promoter, forward primer" promoter complement(2244..2262) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2283..2299) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2283..2299) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2296..2318) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2307..2323 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2331..2361) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2376..2397 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2514..2531) /label=L4440 /note="L4440 vector, forward primer" misc_feature 2636..2660 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" rep_origin complement(2751..3339) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2831..2850) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3510..4325) /codon_start=1 /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" CDS complement(3510..4325) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 4233..4252 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 4317..4336 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 4616..5051 /label=pSa ori /note="origin of replication from bacterial plasmid pSa" misc_feature 5178..5200 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA (truncated)" primer_bind 5350..5372 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5364..5381 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5365..5381 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 5391..5410 /label=T7 /note="T7 promoter, forward primer" promoter 5391..5409 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 5434..5450 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 5435..5451 /label=KS primer /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 catggagtca aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca 61 gttcatacag agtctcttac gactcaatga caagaagaaa atcttcgtca acatggtgga 121 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc 181 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc 241 tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca 301 ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg 361 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca 421 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc 481 gcaagaccct tcctctatat aaggaagttc atttcatttg gagagaacac gggggactct 541 tgacagtggt ctctgtccag tcctggcctc gtcggccatt ctcgactaag ttggcagcat 601 cacccgacgc actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa 661 taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag 721 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg 781 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaactt ttgctgacga 841 gaacagggac tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc 901 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc tgggcgacgg atggtgatcc 961 ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc 1021 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtatccg 1081 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta 1141 acctgatgtt ctggggaata taaatgtcag gctcccttat acacagggcc agtctggcca 1201 cttgtggtct gctgagaccg atggcctcct ccgaggacgt catcaaggag ttcatgcgct 1261 tcaaggtgcg catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg 1321 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc 1381 tgcccttcgc ctgggacatc ctgtcccctc agttccagta cggctccaag gcctacgtga 1441 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg 1501 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc 1561 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc 1621 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcggatg taccccgagg 1681 acggcgccct gaagggcgag atcaagatga ggctgaagct gaaggacggc ggccactacg 1741 acgccgaggt caagaccacc tacatggcca agaagcccgt gcagctgccc ggcgcctaca 1801 agaccgacat caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt 1861 acgagcgcgc cgagggccgc cactccaccg gcgcctgagg tgaccagctc gaatttcccc 1921 gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg 1981 atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc 2041 atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac 2101 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct 2161 atgttactag atcgggcctg cagcccgggg gatccactag ttctagagcg gccgccaccg 2221 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca 2281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 2341 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 2401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 2461 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 2521 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 2581 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgaa ggccttgaca 2641 ggatatattg gcgggtaaac taagtcgctg tatgtgtttg tttgagatct catgtgagca 2701 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 2761 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 2821 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2881 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2941 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 3001 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 3061 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 3121 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 3181 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaga 3241 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 3301 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 3361 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 3421 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 3481 agtatatatg tgtaacattg gtctagtgat tagaaaaact catcgagcat caaatgaaac 3541 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat 3601 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg 3661 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta 3721 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc 3781 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca 3841 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg 3901 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca 3961 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct 4021 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc 4081 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg 4141 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat 4201 cggtagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa 4261 tcagcatcca tgttggaatt taatcgcggc cttgagcaag acgtttcccg ttgaatatgg 4321 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat 4381 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga 4441 ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac 4501 cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc 4561 atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc gatccccatc 4621 caacagcccg ccgtcgagcg ggctttttta tccccggaag cctgtggata gagggtagtt 4681 atccacgtga aaccgctaat gccccgcaaa gccttgattc acggggcttt ccggcccgct 4741 ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg aaatcgctaa tcggagtacg 4801 tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa aggcacgtga gaacgctaat 4861 agccctttca gatcaacagc ttgcaaacac ccctcgctcc ggcaagtagt tacagcaagt 4921 agtatgttca attagctttt caattatgaa tatatatatc aattattggt cgcccttggc 4981 ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga caaccgcaag cggttgccca 5041 ccgtcgagcg cctttgccca caacccggcg gccggccgca acagatcgtt ttataaattt 5101 ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc tcgggcttct ggatttccga 5161 tccccggaat tagatcttgg caggatatat tgtggtgtaa cgttaacatt aacgcttaca 5221 atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt 5281 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc 5341 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact 5401 cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc gata //

pM5MF529

17686

Low

David Shalloway

Lin et al Oncogene. 1995 Jan 19. 10(2):401-5.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/251712/1a871fbb-6fc0-48ce-803f-35a843b80509/addgene-plasmid-17686-sequence-251712.gbk

full

LOCUS sequence-251712- 9448 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9448) AUTHORS Lin PH, Shenoy S, Galitski T, Shalloway D TITLE Transformation of mouse cells by wild-type mouse c-Src. JOURNAL Oncogene. 1995 Jan 19. 10(2):401-5. PUBMED 7530829 REFERENCE 2 (bases 1 to 9448) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Oncogene. 1995 Jan 19. 10(2):401-5." FEATURES Location/Qualifiers source 1..9448 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 1..470 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 535..892 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" primer_bind complement(981..1000) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" rep_origin complement(5995..6583) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6075..6094) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6754..7614) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7377..7396 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7615..7719) /gene="bla" /label=AmpR promoter primer_bind 7787..7805 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa 61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc 121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag 181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg 241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat 301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg 361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct 421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct 481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg 541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct 601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga 661 gttcggaaca cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg 721 tggcccgacc tgagtccaaa aatcccgatc gttttggact ctttggtgca ccccccttag 781 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg 841 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcaggcgca 901 gaactggtag gtatggaaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc 961 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg 1021 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg 1081 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg 1141 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaca 1201 gcacacaagg tagtttcacc aaaccaaaag cgctaccacc cgcaattttc ctcatttgtg 1261 gggatcgcgc atggcaagga attcccagtc gtccggtagg gggcccctgc tatttaggca 1321 agcttaccat gttagcacct aagcatacag atattctcaa ggtgcttgtc aattcatcgc 1381 ggacaggtat aagacgtaaa cgaagcacct cacacctgga tgatacatgc tcagatgaag 1441 tgcagctttg gggtcctaca gcaagaatct ttgcatctat cctagccccg ggggtagcag 1501 ctgcgcaagc cttaagagaa attgagagac tagcctgttg gtccgttaaa caggctaact 1561 tgacaacatc actcctcggg gacttattgg atgatgtcac gagtattcga cacgcggtcc 1621 tgcagaaccg agcggctatt gacttcttgc tcctagctca cggccatggc tgtgaggacg 1681 ttgccggaat gtgctgtttc aatttgagtg atcacagtga gtctatacag aagaagttcc 1741 agctaatgaa ggaacatgtc aataagatcg gcgtggacag cgacccaatt ggaagttggc 1801 tgcgaggact attcggggga ataggagaat gggccgttca tttgctgaaa ggactgcttt 1861 tggggcttgt agttattttg ttgctagtag tgtgcctgcc ttgccttttg caaatcgtat 1921 gcggtaacat cagaaagatg attaataact ccatcagcta ccacacggaa tataagaagc 1981 tgcaaaaggc ctgtgggcag cctgaaagca gaatagtata aggcagtaca tgggtggtgg 2041 tatagcgctt gcgagtcggg ttgtaacggg acatggctta actaagggga ctatggcatg 2101 tataggcgca aagcggggtt tcggttgtac gcgattagga gtccccttag gatatagtag 2161 acacgctttt gcatatgata cataacttcc ctgttttgcc cttagactat tcaagttgcc 2221 tctgtggatt agggctggag gcagctcgga tggtctgacg gccagataag gcaggaaaga 2281 cagctattgg taattgtgaa atacgctttt gtctgtgtgc tgcaggagct gagctgactc 2341 tgctggtggc ctcgcgtacc actgtggcca ggcggtagct gggacgtgca gccgaccacc 2401 atgggcagca acaagagcaa gcccaaggac gccagccagc ggcgccgcag cctggagccc 2461 tcggaaaacg tgcacggggc agggggcgcc ttcccggcct cacagacacc gagcaagccc 2521 gcctccgccg acggccaccg cgggcccagc gccgccttcg tgccgcccgc ggccgagccc 2581 aagctcttcg gaggcttcaa ctcctcggac accgtcacct ccccgcagag ggcggggcct 2641 ctggcaggtg gggtgaccac ctttgtggcc ctctatgact atgagtcacg gacagagact 2701 gacctgtcct tcaagaaagg ggagcggctg cagattgtta acaacacgga gggagactgg 2761 tggctggcac actcgctgag cacgggacag accggttaca tccccagcaa ctatgtggcg 2821 ccctccgact ccatccaggc tgaggagtgg tactttggca agatcactag acgggaatca 2881 gagcggctgc tgctcaacgc cgagaacccg agagggacct tcctcgtgag ggagagtgag 2941 accacaaaag gtgcctactg cctctctgta tccgacttcg acaatgccaa gggcctaaat 3001 gtgaaacact acaagatccg caagctggac agcggcggtt tctacatcac ctcccgcacc 3061 cagttcaaca gcctgcagca gctcgtggct tactactcca aacatgctga tggcctgtgt 3121 caccgcctca ctaccgtatg tcccacatcc aagcctcaga cccagggatt ggccaaggat 3181 gcgtgggaga tcccccggga gtccctgcgg ctggaggtca agctgggcca gggttgcttc 3241 ggagaggtgt ggatggggac ctggaacggc accacgaggg ttgccatcaa aactctgaag 3301 ccaggcacca tgtccccaga ggccttcctg caggaggccc aagtcatgaa gaaactgagg 3361 cacgagaaac tggtgcagct gtatgctgtg gtgtcggaag aacccattta cattgtgaca 3421 gagtacatga acaaggggag tctgctggac tttctcaagg gggaaacggg caaatatttg 3481 cggctacccc agctggtgga catgtctgct cagatcgctt caggcatggc ctatgtggag 3541 cggatgaact atgtgcaccg ggaccttcga gccgccaata tcctagtagg ggagaacctg 3601 gtgtgcaaag tggccgactt tgggttggcc cggctcatag aagacaacga atacacagcc 3661 cggcaaggtg ccaaattccc catcaagtgg acagcccccg aggcagccct ctatggccgg 3721 ttcaccatca agtcggatgt ctggtccttc ggcatcctgc tgactgagct gaccaccaag 3781 ggccgggtgc catacccagg tgagagttag catcaacccc acaccccgtg ggttgggcac 3841 agcgtgtgac ccggccccgc tgcccttctt tccacaggga tggtcaacag ggaggtgctg 3901 gaccaggtgg agaggggcta ccgcatgccc tgcccgcccg agtgccccga gtcgctgcat 3961 gacctcatgt gccagtgctg gcggaaggac cctgaggagc ggcccacttt tgagtacctg 4021 caggccttcc tggaggacta cttcacctcg acagagcccc agttccagcc tggagagaac 4081 ctatagatct ctagaagctt atcgattagt ccaatttgtt aaagacagga tatcagtggt 4141 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata 4201 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg 4261 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact 4321 gagaatagaa aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa 4381 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca 4441 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa 4501 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg 4561 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc 4621 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt 4681 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc 4741 cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa 4801 gtatttacat taaatggcca tagtacttaa agttacattg gcttccttga aataaacatg 4861 gagtattcag aatgtgtcat aaatatttct aattttaaga tagtatctcc attggctttc 4921 tactttttct tttatttttt tttgtcctct gtcttccatt tgttgttgtt gttgtttgtt 4981 tgtttgtttg ttggttggtt ggttaatttt tttttaaaga tcctacacta tagttcaagc 5041 tagactatta gctactctgt aacccagggt gaccttgaag tcatgggtag cctgctgttt 5101 tagccttccc acatctaaga ttacaggtat gagctatcat ttttggtata ttgattgatt 5161 gattgattga tgtgtgtgtg tgtgattgtg tttgtgtgtg tgattgtgta tatgtgtgta 5221 tggttgtgtg tgattgtgtg tatgtatgtt tgtgtgtgat tgtgtgtgtg tgattgtgca 5281 tgtgtgtgtg tgtgattgtg tttatgtgta tgattgtgtg tgtgtgtgtg tgtgtgtgtg 5341 tgtgtgtgtg tgtgtgtgtg ttgtgtatat atatttatgg tagtgagagg caacgctccg 5401 gctcaggtgt caggttggtt tttgagacag agtctttcac ttagcttgga attctcatgt 5461 ttgacagctt atcatcgatg ataagctgtc aaacatgaga atttcgaccg atgcccttga 5521 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac 5581 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca 5641 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat 5701 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg 5761 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg 5821 cgttccgacg cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat 5881 cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca 5941 gcttcaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 6001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 6061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 6121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 6181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6421 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 6481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 6541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 6601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 6661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 6721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 6781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 6841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 6901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 6961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 7021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt 7081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 7141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 7201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa 7381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 7441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 7501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 7561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 7621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 7681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 7741 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 7801 gaggcccttt cgtcttcaag aattcattct tttattgagt ccagattaga aaaagaagag 7861 aatttatgaa cggacctttt gacagatttc tttttgaaaa tactcattcc tcttcagatt 7921 tgttttaaat gaagttttct ttctttaact tgtttgcacg cagctcagct gtgttctttc 7981 ttaaatgagg tctggtaaag aaaaggccct accaaggtct tagctgtact tgtgacatga 8041 tctctctatt aggttaaagt ttttttctgt tatgtgatgc tcataaaact gagttataac 8101 aatattaata ataacttcac taattgattc tgtgagggat gagaagaggg aggaatagga 8161 tgagaaaaac agagggaact atttgccttg tatttgacat acttagaaga taagggaaac 8221 atgtttggaa tgtcagttcc ccaggctact ttatgtcaat gttagaaatg gaaacaatga 8281 ttttcattgg ctacaaacag gagaggaggc tgtggtgatt ttggggtatg cgttgtgttc 8341 attgtgattc tataagcacc ccatcttcca aagagagctc aacaatagtc aggaagcctc 8401 gccctaaagc ccatcctccc ccccgcccca cccccatgac tttgtttcct tgtcagactt 8461 tacctgagag gaaatgaaat taataagctg tgaaaaagta tcccatgtgg gctgctctta 8521 agtctcccgt tgccttttcc tcctgtgttc tgcgttctag ctgtttgcga gtaggtgagg 8581 ctggtgactc actgccttca agcagcagca ggactggtga gtcatagctt tcaggctggg 8641 gttgtggttc actcccatgg acggtttaac ccgtggttgt ctcttagggg gtcagtggga 8701 acatccttcg tatggtcgtg atcttcttaa atcctctccc caggaccagg caataacagt 8761 gtcctggcaa aagctccagt actccatctt ctcaggacat cccagtccac atgactaatc 8821 tttcccacct tctacatttg agcgtttcct gccttcacat tatagaaact attttgggaa 8881 gtattatttt taaacatgac ttttccctta attctaccat aataagtgct gggttaagct 8941 ttgctcttag gagtttccta atacatccca aactcaaata tataaagcat ttgacttgtt 9001 ctatgcccta gggggcgggg ggaagctaag ccagcttttt ttaacattta aaatgttaat 9061 tccattttaa atgcacagat gtttttattt cataagggtt tcaatgtgca tgaatgctgc 9121 aatattcctg ttaccaaagc tagtataaat aaaaatagat aaacgtggaa attacttaga 9181 gtttctgtca ttaacgtttc cttcctcagt tgacaacata aatgcgctgc tgagaagcca 9241 gtttgcatct gtcaggatca atttcccatt atgccagtca tattaattac tagtcaatta 9301 gttgattttt atttttgaca tatacatgtg aaagacccca cctgtaggtt tggcaagcta 9361 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agaaaagttc 9421 agatcaaggt caggaacaga tggaacag //

pAc5.1B-DmAdhV5-intronless-5BoxB_X

147911

Insect Expression of DmAdh-intronless-5BoxB

Elisa Izaurralde

Izaurralde lab plasmids (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/290358/dba0a908-f5e3-4c44-b6e8-7d110e2d1f03/addgene-plasmid-147911-sequence-290358.gbk

full

LOCUS sequence-290358- 6433 bp DNA circular SYN 26-AUG-2024 DEFINITION Insect Expression of DmAdh-intronless-5BoxB. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6433) TITLE Izaurralde lab plasmids REFERENCE 2 (bases 1 to 6433) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..6433 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 56..2568 /label=Ac5 promoter /note="Drosophila melanogaster actin 5C promoter" CDS 3378..3419 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" protein_bind 3504..3522 /label=boxB /bound_moiety="lambda-N peptide" /note="lambda N peptide binding site consisting of an RNA hairpin within the N-utliization (nut) site of the lambda bacteriophage transcript (Tan and Frankel, 1995)" protein_bind 3546..3564 /label=boxB /bound_moiety="lambda-N peptide" /note="lambda N peptide binding site consisting of an RNA hairpin within the N-utliization (nut) site of the lambda bacteriophage transcript (Tan and Frankel, 1995)" protein_bind 3570..3588 /label=boxB /bound_moiety="lambda-N peptide" /note="lambda N peptide binding site consisting of an RNA hairpin within the N-utliization (nut) site of the lambda bacteriophage transcript (Tan and Frankel, 1995)" protein_bind 3628..3646 /label=boxB /bound_moiety="lambda-N peptide" /note="lambda N peptide binding site consisting of an RNA hairpin within the N-utliization (nut) site of the lambda bacteriophage transcript (Tan and Frankel, 1995)" protein_bind 3652..3670 /label=boxB /bound_moiety="lambda-N peptide" /note="lambda N peptide binding site consisting of an RNA hairpin within the N-utliization (nut) site of the lambda bacteriophage transcript (Tan and Frankel, 1995)" CDS 3718..3759 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 3769..3786 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3809..3826) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3859..3993 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3883..3902) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3937..3956 /label=SV40pA-R /note="SV40 polyA, reverse primer" rep_origin complement(4615..5203) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4695..4714) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5374..6234) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5997..6016 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6235..6339) /gene="bla" /label=AmpR promoter primer_bind 6407..6425 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga 61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac 121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg 181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga 241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg 301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc 361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac 421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct 481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag 541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt 601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac 661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt 721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg 781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct 841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca 901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg 961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg 1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc 1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg 1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa 1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa 1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag 1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa 1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt 1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag 1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta 1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga 1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc 1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc 1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca 1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat 1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta 1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag 1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta 2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac 2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa 2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga 2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca 2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa 2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact 2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg 2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct 2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg 2581 tacctactag tccagtgtgg tggaattcca acatgtcgtt tactttgacc aacaagaacg 2641 tgattttcgt tgccggtctg ggaggcattg gtctggacac cagcaaggag ctgctcaagc 2701 gcgatctgaa gaacctggtg atcctcgacc gcattgagaa cccggctgcc attgccgagc 2761 tgaaggcaat caatccaaag gtgaccgtca ccttctaccc ctatgatgtg accgtgccca 2821 ttgccgagac caccaagctg ctgaagacca tcttcgccca gctgaagacc gtcgatgtcc 2881 tgatcaacgg agctggtatc ctggacgatc accagatcga gcgcaccatt gccgtcaact 2941 acactggcct ggtcaacacc acgacggcca ttctggactt ctgggacaag cgcaagggcg 3001 gtcccggtgg tatcatctgc aacattggat ccgtcactgg attcaatgcc atctaccagg 3061 tgcccgtcta ctccggcacc aaggccgccg tggtcaactt caccagctcc ctggcgaaac 3121 tggcccccat taccggcgtg accgcttaca ccgtgaaccc cggcatcacc cgcaccaccc 3181 tggtgcacaa gttcaactcc tggttggatg ttgagcccca ggttgctgag aagctcctgg 3241 ctcatcccac ccagccatcg ttggcctgcg ccgagaactt cgtcaaggct atcgaactga 3301 accagaacgg agccatctgg aaactggact tgggcaccct ggaggccatc cagtggacca 3361 agcactggga ctccggcggt aagcctatcc ctaaccctct cctcggtctc gattctacgt 3421 aaatctcgag ggaattcgct agcttccccg ctcgagggaa ttcgctagct tccctaagtc 3481 caactaccaa actggggatt cctgggccct gaagaagggc ccctcgacta gtccaaatac 3541 aaactgggcc ctgaagaagg gcccatatag ggccctgaag aagggcccta tcgaggatat 3601 tatctcgact aagtccaact acaaactggg ccctgaagaa gggcccatat agggccctga 3661 agaagggccc tatcgaggat attatctcga agtcacccat tcgaaccgcg gttcgaaggt 3721 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat 3781 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc 3841 tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt tggacaaacc 3901 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta 3961 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg 4021 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt 4081 ggtatggctg attatgatca gtcgaccgat gcccttgaga gccttcaacc cagtcagctc 4141 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat 4201 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg 4261 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc 4321 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc 4381 cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat 4441 ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc 4501 catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc 4561 tcttaccagc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 4621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 4681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 4741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 4801 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 4861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 4921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 4981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 5041 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 5101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 5161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 5221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 5281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 5341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 5401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 5461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 5521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 5581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 5641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt 5701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 5761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 5821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 5881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 5941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa 6001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 6061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 6121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 6181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 6241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 6301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 6361 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 6421 gaggcccttt cgt //

pLXMA L1A

153347

(Empty Backbone)Expression vector for plants with medium-copy number origin of replication

Naomi Nakayama

Andreou et al bioRxiv, March 31, 2021

https://media.addgene.org/snapgene-media/v2.0.0/sequences/414833/64c52f10-cb85-4013-8e61-1686ab0547ed/addgene-plasmid-153347-sequence-414833.gbk

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LOCUS sequence-414833- 3799 bp DNA circular SYN 26-AUG-2024 DEFINITION Expression vector for plants with medium-copy number origin of replication. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3799) AUTHORS Andreou , Andreas I., Nirkko, Jessica , Ochoa-Villarreal, Marisol , Nakayama, Naomi TITLE Mobius Assembly for Plant Systems highlights promoter-terminator interaction in gene regulation JOURNAL bioRxiv, March 31, 2021 REFERENCE 2 (bases 1 to 3799) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv, March 31, 2021" FEATURES Location/Qualifiers source 1..3799 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 723..747 /label=LB T-DNA repeat /note="left border repeat from octopine Ach5 T-DNA" misc_feature 774..798 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" promoter 840..874 /label=BBa_J23106 /note="Anderson constitutive promoter (https://parts.igem.org/Promoters/Catalog/Anderson). Measured strength relative to BBa_J223100: 0.47" RBS 883..894 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" terminator 1583..1654 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1670..1697 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(1927..2742) /codon_start=1 /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 2734..2753 /label=pENTR-R /note="pENTR vectors, reverse primer" CDS complement(2868..3530) /codon_start=1 /product="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" /label=pBBR1 Rep /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR" rep_origin join(3531..3799,1..503) /label=pBBR1 oriV /note="replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication" ORIGIN 1 ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga 61 tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact 121 cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc ccttcgactg cccccactcg 181 cataggcttg ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag 241 ccttgacccg ccttccactt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg 301 gaggcttttc cccagagaaa attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt 361 tggagccggt gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag 421 ctcgtgggca ggcgcagcct gtccatcagc ttgtccagca gggttgtcca cgggccgagc 481 gaagcgagcc agccggtggc cgctcgcggc catcgtccac atatccacgg gctggcaagg 541 gagcgcagcg accgcgcagg gcgaagcccg gagagcaagc ccgtaggggg ccatcatcag 601 ttcggtggtc ttccgacgaa caataaggcc gcaaatcgcg gcctttttta ttgataacaa 661 aaccggctca gttctgcgta gaaaccaaca tgcaagctcc accgggtgca aagcggcagc 721 ggcggcagga tatattcaat tgtaaatggc ttcatgtccg ggaaatctac tggtggcagg 781 atatattgtg gtgtaaacaa tggagaaaaa gagaattcac ctgcatatgg agagagacct 841 ttacggctag ctcagtccta ggtatagtgc tagctactag agaaagagga gaaatactaa 901 atgtcgcact caaaacaagc actggcagat acgatgaaaa tgacgtggct gatggaaggt 961 tcggtcaacg gtcacgcctt tacgattgaa ggcgaaggta ccggcaaacc gtatgaaggt 1021 aaacagagtg gcacctttcg tgttacgaaa ggcggtccgc tgccgtttgc gtttgatatt 1081 gtcgctccga ccctgaaata cggtttcaaa tgcttcatga aatacccggc ggatattccg 1141 gactacttta aactggcctt cccggaaggc ctgacgtacg atcgtaaaat tgcgtttgaa 1201 gacggcggtt gtgcgaccgc cacggtcgaa atgagcctga agggtaacac cctggtgcat 1261 aaaacgaact ttcagggcgg taatttcccg attgatggtc cggtgatgca aaaacgcacc 1321 ctgggctggg aaccgacctc tgaaaaaatg acgccgtgcg atggtattat caaaggcgac 1381 accatcatgt acctgatggt tgaaggcggt aaaacgctga aatgtcgtta tgaaaacaat 1441 taccgcgcca acaaaccagt gctgatgccg ccgagccact ttgtggatct gcgtctgacc 1501 cgcacgaatc tggataaaga aggtctggcg ttcaaactgg aagaatatgc tgttgcccgt 1561 gtgctggaag tgtaataagc ttcaaataaa acgaaaggct cagtcgaaag actgggcctt 1621 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc 1681 gggtgggcct ttctgcgggt ctcacgctca gaatatgcag gtgctgcagg aattgttgat 1741 tttgtgatga ctgatggcag gatatatgcg gttgtaattc atttttattg tctaaatttc 1801 tgtatttgtt tgtttgttcg gttgtaaatt tttttggaag aacaagaaaa gaaaaaacac 1861 ccgttagggt gtttttagtt agtgtggcgc gccgacttgc gacatgcggt cctttgcaat 1921 caactattag aaaaattcat ccagcatcag atgaaattgc agtttgttca tatccggatt 1981 atcaatgcca tatttctgaa acagacgttt ttgcaggctc gggctaaatt cgcccaggca 2041 gttccacaga atggccagat cctgataacg atccgcaatg cccacacggc ccacatcaat 2101 gcagccaatc agtttgcctt catcgaaaat caggttatcc aggctaaaat cgccgtgggt 2161 caccacgcta tccgggctaa acggcagcag tttatgcatt tctttccaca cctgttccac 2221 cggccagccg ttacgttcat catcaaaatc gctcgcatcc accaggccgt tgttcatacg 2281 gctctgcgcc tgggccagac gaaacacacg atcgctgtta aacgggcagt tgcacaccgg 2341 aatgctatgc agacgacgca gaaacacggc cagcgcatcc acaatgtttt cgccgctatc 2401 cggatattct tccagcacct gaaacgcggt tttgcccgga atcgcggtgg tcagcagcca 2461 cgcatcatcc ggggtgcgaa taaaatgttt aatggtcggc agcggcataa attcggtcag 2521 ccagttcaga cgcaccattt catcggtcac atcgttcgcc acgctgcctt tgccatgttt 2581 cagaaacagt tccggcgcat ccggtttgcc atacagacga taaatggtcg cgccgctctg 2641 acccacgtta tcacgcgccc atttatagcc atacagatcc gcatccatgt tgctgttcag 2701 acgcggacgg ctacagctcg tttcacgctg aatatggctc ataacacccc ttgtattact 2761 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta 2821 acatcagaga ttttgagaca caagatcgga ttggcggtta tgcggttcta ccggcgcggc 2881 agcgttaccc gtgtcggcgg ctccaacggc tcgccatcgt ccagaaaaca cggctcatcg 2941 ggcatcggca ggcgctgctg cccgcgccgt tcccattcct ccgtttcggt caaggctggc 3001 aggtctggtt ccatgcccgg aatgccgggc tggctgggcg gctcctcgcc ggggccggtc 3061 ggtagttgct gctcgcccgg atacagggtc gggatgcggc gcaggtcgcc atgccccaac 3121 agcgattcgt cctggtcgtc gtgatcaacc accacggcgg cactgaacac cgacaggcgc 3181 aactggtcgc ggggctggcc ccacgccacg cggtcattga ccacgtaggc cgacacggtg 3241 ccggggccgt tgagcttcac gacggagatc cagcgctcgg ccaccaagtc cttgactgcg 3301 tattggaccg tccgcaaaga acgtccgatg agcttggaaa gtgtcttctg gctgaccacc 3361 acggcgttct ggtggcccat ctgcgccacg aggtgatgca gcagcattgc cgccgtgggt 3421 ttcctcgcaa taagcccggc ccacgcctca tgcgctttgc gttccgtttg cacccagtga 3481 ccgggcttgt tcttggcttg aatgccgatt tctctggact gcgtggccat gcttatctcc 3541 atgcggtagg ggtgccgcac ggttgcggca ccatgcgcaa tcagctgcaa cttttcggca 3601 gcgcgacaac aattatgcgt tgcgtaaaag tggcagtcaa ttacagattt tctttaacct 3661 acgcaatgag ctattgcggg gggtgccgca atgagctgtt gcgtaccccc cttttttaag 3721 ttgttgattt ttaagtcttt cgcatttcgc cctatatcta gttctttggt gcccaaagaa 3781 gggcacccct gcggggttc //

pMIR-His-Halo-Tev-PTEN

58241

expresses halo tagged PTEN protein

Yimon Aye

Fang et al J Am Chem Soc. 2013 Oct 2;135(39):14496-9. doi: 10.1021/ja405400k. Epub 2013 Sep 18.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/175309/44f0df21-7123-4a6c-a057-26dee2ea0384/addgene-plasmid-58241-sequence-175309.gbk

full

LOCUS sequence-175309- 6876 bp DNA circular SYN 25-AUG-2024 DEFINITION expresses halo tagged PTEN protein. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6876) AUTHORS Fang X, Fu Y, Long MJ, Haegele JA, Ge EJ, Parvez S, Aye Y TITLE Temporally controlled targeting of 4-hydroxynonenal to specific proteins in living cells. JOURNAL J Am Chem Soc. 2013 Oct 2;135(39):14496-9. doi: 10.1021/ja405400k. Epub 2013 Sep 18. PUBMED 24015839 REFERENCE 2 (bases 1 to 6876) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1021/ja405400k"; journalName: "J Am Chem Soc"; date: "2013-10-2- 2"; volume: "135"; issue: "39"; pages: "14496-9" FEATURES Location/Qualifiers source 1..6876 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 44..643 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(44..63) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 540..560 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 873..954 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(910..929) /label=SV40pA-R /note="SV40 polyA, reverse primer" promoter 1042..1146 /gene="bla" /label=AmpR promoter CDS 1147..2007 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1365..1384) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2178..2766 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2667..2686 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2920..2937 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3054..3075 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3090..3120 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3128..3144 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 3133..3155 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3152..3168 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3152..3168 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 3188..3206 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 3188..3205 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 3218..3237 /label=T7 /note="T7 promoter, forward primer" promoter 3218..3236 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" enhancer 3358..3661 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3662..3865 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3816..3836 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 3902..3919 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 3929..3946 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 3950..4840 /codon_start=1 /product="modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives" /label=HaloTag(R) /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL IGSEIARWLSTLEISG" CDS 4847..4867 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" primer_bind complement(6101..6118) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6101..6117) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6110..6132) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(6326..6345) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6445..6467 /label=pGEX 3' /note="pGEX vectors, reverse primer" promoter 6507..6823 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6674..6809 /label=SV40 ori /note="SV40 origin of replication" ORIGIN 1 ccctgaccca cgcccctgac ccctcacaag gagacgacct tccatgaccg agtacaagcc 61 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc 121 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt 181 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt 241 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc 301 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca 361 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc 421 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc 481 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg 541 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga 601 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc 661 gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc acccggggcg 721 gccccgccga ccccgcaccc gcccccgagg cccaccgact ctagaggatc ataatcagcc 781 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc 841 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt 901 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcaatctc 961 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 1021 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 1081 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 1141 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 1201 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 1261 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 1321 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 1381 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 1441 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 1501 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 1561 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 1621 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 1681 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 1741 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 1801 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 1861 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 1921 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 1981 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 2041 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 2101 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 2161 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 2221 aaaaaaccac cgctaccagc ggtggttttt ttgccggatc aagagctacc aactcttttt 2281 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 2341 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 2401 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 2461 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 2521 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 2581 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 2641 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 2701 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 2761 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 2821 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 2881 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 2941 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 3001 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 3061 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 3121 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat 3181 tgcaacgatt taggtgacac tatagaagag aaggaattaa tacgactcac tatagggaga 3241 gagagagaat taccctcact aaagggagga gaagcatgaa ttcaaggtac cagatcttag 3301 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt 3361 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 3421 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg 3481 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 3541 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat 3601 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat 3661 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt 3721 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga 3781 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg 3841 gtgggaggtc tatataagca gagctggttt agtgaaccgt ggatccacca tgggcagcag 3901 ccatcatcat catcatcaca gcagcggcct ggtgcctcgt ggtagccata tggcagaaat 3961 cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta 4021 cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc 4081 ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc 4141 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca 4201 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat 4261 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa 4321 aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt 4381 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga 4441 tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt 4501 cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg 4561 cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga 4621 atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg 4681 cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc 4741 tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag 4801 cgagatcgcg cgctggctgt cgacgctcga gatttccggc tccggagaaa acttgtattt 4861 ccagggctca gggatgacag ccatcatcaa agagatcgtt agcagaaaca aaaggagata 4921 tcaagaggat ggattcgact tagacttgac ctatatttat ccaaacatta ttgctatggg 4981 atttcctgca gaaagacttg aaggcgtata caggaacaat attgatgatg tagtaaggtt 5041 tttggattca aagcataaaa accattacaa gatatacaat ctttgtgctg aaagacatta 5101 tgacaccgcc aaatttaatt gcagagttgc acaatatcct tttgaagacc ataacccacc 5161 acagctagaa cttatcaaac ccttttgtga agatcttgac caatggctaa gtgaagatga 5221 caatcatgtt gcagcaattc actgtaaagc tggaaaggga cgaactggtg taatgatatg 5281 tgcatattta ttacatcggg gcaaattttt aaaggcacaa gaggccctag atttctatgg 5341 ggaagtaagg accagagaca aaaagggagt aactattccc agtcagaggc gctatgtgta 5401 ttattatagc tacctgttaa agaatcatct ggattataga ccagtggcac tgttgtttca 5461 caagatgatg tttgaaacta ttccaatgtt cagtggcgga acttgcaatc ctcagtttgt 5521 ggtctgccag ctaaaggtga agatatattc ctccaattca ggacccacac gacgggaaga 5581 caagttcatg tactttgagt tccctcagcc gttacctgtg tgtggtgata tcaaagtaga 5641 gttcttccac aaacagaaca agatgctaaa aaaggacaaa atgtttcact tttgggtaaa 5701 tacattcttc ataccaggac cagaggaaac ctcagaaaaa gtagaaaatg gaagtctatg 5761 tgatcaagaa atcgatagca tttgcagtat agagcgtgca gataatgaca aggaatatct 5821 agtacttact ttaacaaaaa atgatcttga caaagcaaat aaagacaaag ccaaccgata 5881 cttttctcca aattttaagg tgaagctgta cttcacaaaa acagtagagg agccgtcaaa 5941 tccagaggct agcagttcaa cttctgtaac accagatgtt agtgacaatg aacctgatca 6001 ttatagatat tctgacacca ctgactctga tccagagaat gaaccttttg atgaagatca 6061 gcatacacaa attacaaaag tctgaggccg ctagcttggc actggccgtc gttttacaac 6121 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 6181 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 6241 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt 6301 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc 6361 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg 6421 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat 6481 caccgaaacg cgcgagacga aagattggtg tggaaagtcc ccaggctccc cagcaggcag 6541 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 6601 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 6661 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 6721 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 6781 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctag cttgcatgcc 6841 tgcaggtcgg ccgccacgac cggtgccgcc accatc //

pMSR:MSMEG_0777:Dendra

174913

C-term fluorescent fusion

Judd et al mBio. 2021 Mar 2;12(2). pii: mBio.02401-20. doi: 10.1128/mBio.02401-20.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/345462/9e3db63d-0347-46ce-9dad-891d76669db2/addgene-plasmid-174913-sequence-345462.gbk

full

LOCUS sequence-345462- 5888 bp DNA circular SYN 26-AUG-2024 DEFINITION C-term fluorescent fusion. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5888) AUTHORS Judd JA, Canestrari J, Clark R, Joseph A, Lapierre P, Lasek-Nesselquist E, Mir M, Palumbo M, Smith C, Stone M, Upadhyay A, Wirth SE, Dedrick RM, Meier CG, Russell DA, Dills A, Dove E, Kester J, Wolf ID, Zhu J, Rubin ER, Fortune S, Hatfull GF, Gray TA, Wade JT, Derbyshire KM TITLE A Mycobacterial Systems Resource for the Research Community. JOURNAL mBio. 2021 Mar 2;12(2). pii: mBio.02401-20. doi: 10.1128/mBio.02401-20. PUBMED 33653882 REFERENCE 2 (bases 1 to 5888) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "mBio."; date: "2021-03-2"; volume: "12(2). pii: mBio.02401-20. doi"; pages: " 10.1128/mBio.02401-20" FEATURES Location/Qualifiers source 1..5888 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 712..731 /label=T7 /note="T7 promoter, forward primer" promoter 712..730 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2498..2521 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 2778..3578 /codon_start=1 /gene="aac(3)IV" /product="aminoglycoside 3-N-acetyltransferase type IV " /label=ApmR /note="confers resistance to apramycin (Paget and Davies, 1996)" /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP LEDGPLGLIEALRATLGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV KRSAHPFAFAAAGPQAEQIISDPLPLPPYSPASPVARVHELDGQVLLLGVGHDANTTLH LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG" rep_origin 3703..4291 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4192..4211 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ggtgaccacg acgcagggca accggctgtc gaagtccgcg ttcaccaagt cgctgaagcg 61 tggctacgcc aagatcggtc ggccggaact ccgcatccac gacctccgcg ctgtcggcgc 121 tacgttcgcc gctcaggcag gtgcgacgac caaggagctg atggcccgtc tcggtcacac 181 gactcctagg atggcgatga agtaccagat ggcgtctgag gcccgcgacg aggctatcgc 241 tgaggcgatg tccaagctgg ccaagacctc ctgaaacgca aaaagccccc ctcccaagga 301 cactgagtcc taaagagggg ggtttcttgt cagtacgcga agaaccacgc ctggccgcga 361 gcgccagcac cgccgctctg tgcggagacc tgggcaccag ccccgccgcc gccaggagca 421 ttgccgttcc cgccagaaat ctagacggtg accacaacgc gcccgctttg atcggggacg 481 tcgaaatatt ggatcgtcgg caccgtcacg gccgtgggag gcggcacgat ccgcgacgtg 541 atgatcggcc gcatccccac ggtgctgcgc agtgagctct acgccatccc ggcgttgatc 601 tgtgcgttcg cacgcacagg cccggtgtga gaagggtctc tgacgagcgg gagaactccc 661 tatcagtgat agagtttgtc ctccctatca gtgatagata ggcttaagga gtaatacgac 721 tcactatagg gagaaggagg ccattatggc tgaattgaag ctaggttaca aggcatctgc 781 ggagcagttt gctcctcgcg agttggtcga gttggcggtt ttggccgagt cggcaggtat 841 ggacagtgcg actgtcagtg atcatttcca gccgtggcgt cacgaaggtg ggcatgcgcc 901 gttctcgctg gcgtggatga ccgcggtggg cgagcgcacc aagaacctgg tgctggggac 961 gtcggtgctg acgccgacgt tccggtacaa cccggcggtg atcgcccagg cgttcgcgac 1021 gatgggatgt ctgtatccgg ggcggatctt cctcggtgtg ggcaccggtg aggcgctcaa 1081 cgagatcgcg accgggtatg ccggcgagtg gccggagttc aaggagcggt tcgcgcggct 1141 acgcgaatcg gtgcggttga tgcgtgagct gtggctgggg gaccgggtcg atttcgacgg 1201 cgagtactac cgcaccaagg gcgcgtcgat ctatgacgtg cccgagggtg gcatcccggt 1261 gtacatcgcc gcgggtggtc cggtggtggc caagtacgcc ggccgtgccg gtgacgggtt 1321 catctgcacc tcgggcaagg gtgaggagct ctatgccgag aagctgatcc cggcggtcaa 1381 ggagggtgct gcggccgccg atcgcgacgc cgacgcgatc gaccggatga tcgagatcaa 1441 gatctcctat gacaccgatc ccgagttggc gctggagaac acccggttct gggcgccgct 1501 gtcgttgacc gccgagcaga agcattcgat cgacgatccg atcgagatgg agaaggctgc 1561 cgacgcgttg cccatcgagc aggtcgccaa gcgctggatc gtcgcgtcgg accccgacga 1621 ggcggtcgag aaggtcggcc agtacgtcaa gtgggggctg aaccacctgg tgttccacgc 1681 cccgggccat gaccagcgtc gattcctcga gctgttcaaa cgcgatctgg aaccgcggtt 1741 gcgcaagctg gccaagcttg gtgcggccgc gggcaacacg cctggcatca atctgatcaa 1801 ggaggacatg cgtgtcaagg tgcatatgga gggcaacgtg aacggtcacg ccttcgtgat 1861 cgagggggag ggtaagggta agccttacga agggacccag accgccaacc tcaccgtcaa 1921 ggagggcgcc cctctcccat tctcgtacga tatcctcact accgcggtgc attacgggaa 1981 ccgcgtgttc accaagtacc ccgaggatat ccctgactac ttcaagcaga gcttcccgga 2041 gggatactcc tgggagcgga cgatgacctt cgaagacaag ggtatctgta cgatccgcag 2101 cgacatctcg ctggaagggg actgtttctt ccagaacgtc cgtttcaagg ggaccaactt 2161 tccgccgaac ggcccggtca tgcagaagaa gaccctcaag tgggagccga gcacggagaa 2221 gctgcacgtt cgggacggcc tcctggtcgg caacatcaac atggcgctgc tgctggaagg 2281 cggcggccac tacctatgcg acttcaagac cacgtacaaa gcgaagaagg tcgtccagct 2341 gcccgacgcg cacttcgtgg accaccgcat cgaaatcctg gggaacgatt cggactacaa 2401 caaggtcaag ttgtacgagc acgctgtggc acgctatagc ccgctcccgt cgcaggtctg 2461 gttcgaaggc ggcggcgggg agaacctgta tttccaggac tacaaggacg acgacgacaa 2521 gtgatttaaa taaccccttg gggcctctaa acgggtcttg aggggttttt tgttaactag 2581 cgtacgatcg aattcctgca gctcacggta actgatgccg tatttgcagt accagcgtac 2641 ggcccacaga atgatgtcac gctgaaaatg ccggcctttg aatgggttca tgtgcagctc 2701 catcagcaaa aggggatgat aagtttatca ccaccgacta tttgcaacag tgccgttgat 2761 cgtgctatga tcgactgatg tcatcagcgg tggagtgcaa tgtcgtgcaa tacgaatggc 2821 gaaaagccga gctcatcggt cagcttctca accttggggt tacccccggc ggtgtgctgc 2881 tggtccacag ctccttccgt agcgtccggc ccctcgaaga tgggccactt ggactgatcg 2941 aggccctgcg tgctacgctg ggtccgggag ggacgctcgt catgccctcg tggtcaggtc 3001 tggacgacga gccgttcgat cctgccacgt cgcccgttac accggacctt ggagttgtct 3061 ctgacacatt ctggcgcctg ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg 3121 cagcggggcc acaggcagag cagatcatct ctgatccatt gcccctgcca ccttactcgc 3181 ctgcaagccc ggtcgcccgt gtccatgaac tcgatgggca ggtacttctc ctcggcgtgg 3241 gacacgatgc caacacgacg ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg 3301 tgccgagaca ctgcaccatt cttcaggatg gcaagttggt acgcgtcgat tatctcgaga 3361 atgaccactg ctgtgagcgc tttgccttgg cggacaggtg gctcaaggag aagagccttc 3421 agaaggaagg tccagtcggt catgcctttg ctcggttgat ccgctcccgc gacattgtgg 3481 cgacagccct gggtcaactg ggccgagatc cgttgatctt cctgcatccg ccagaggcgg 3541 gatgcgaaga atgcgatgcc gctcgccagt cgattggctg agctcatgag cggagaacga 3601 gatgacgttg gaggggcaag gtcgcgctga ttgctggggc aacacggggg atccactagt 3661 tccatgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 3721 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 3781 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 3841 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 3901 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 3961 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 4021 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 4081 cctacagcgt gagctatgag aaagcgccac gcttcccgag gggagaaagg cggacaggta 4141 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 4201 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 4261 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 4321 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 4381 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 4441 gcgcaacgcg tgcggccgcg gtacccgggg atcctctaga gtcgaccacc aagggcacca 4501 tctctgcttg ggccaccccg ttggccgcag ccagctcgct gagagccgtg aacgacaggg 4561 cgaacgccag cccgccgacg gcgagggttc cgaccgctgc aactcccggt gcaaccttgt 4621 cccggtctat tctcttcact gcaccagctc caatctggtg tgaatgcccc tcgtctgttc 4681 gcgcaggcgg ggggctctat tcgtttgtca gcatcgaaag tagccagatc agggatgcgt 4741 tgcaaccgcg tatgcccagg tcagaagagt cgcacaagag ttgcagaccc ctggaaagaa 4801 aaatggccag agggcgaaaa caccctctga ccagcggagc gggcgacggg aatcgaaccc 4861 gcgtagctag tttggaagaa tgggtgtctg ccgaccacat atgggccggt caagataggt 4921 ttttaccccc tctcggctgc atcctctaag tggaaagaaa ttgcaggtcg tagaagcgcg 4981 ttgaagcctg agagttgcac aggagttgca acccggtagc cttgttcacg acgagaggag 5041 acctagttgg cacgtcgcgg atggggatcg ctgaagactc agcgcagcgg gaggatccaa 5101 gcctcatacg tcaacccgca ggacggtgtg aggtactacg cgctgcagac ctacgacaac 5161 aagatggacg ccgaagcctg gctcgcgggc gagaagcggc tcatcgagat ggagacctgg 5221 acccctccac aggaccgggc gaagaaggca gccgccagcg ccatcacgct ggaggagtac 5281 acccggaagt ggctcgtgga gcgcgacctc gcagacggca ccagggatct gtacagcggg 5341 cacgcggagc gccgcatcta cccggtgcta ggtgaagtgg cggtcacaga gatgacgcca 5401 gctctggtgc gtgcgtggtg ggccgggatg ggtaggaagc acccgactgc ccgccggcat 5461 gcctacaacg tcctccgggc ggtgatgaac acagcggtcg aggacaagct gatcgcagag 5521 aacccgtgcc ggatcgagca gaaggcagcc gatgagcgcg acgtagaggc gctgacgcct 5581 gaggagctgg acatcgtcgc cgctgagatc ttcgagcact accggatcgc ggcatacatc 5641 ctggcgtgga cgagcctccg gttcggagag ctgatcgagc ttcgccgcaa ggacatcgtg 5701 gacgacggca tgacgatgaa gctccgggtg cgccgtggcg cttcccgcgt ggggaacaag 5761 atcgtcgttg gcaacgccaa gaccgtccgg tcgaagcgtc ctgtgacggt tccgcctcac 5821 gtcgcggaga tgatccgagc gcacatgaag gaccgtacga agatgaacaa gggccccgag 5881 gcattcct //

GRM1-Tango

66387

Medium

Expression of G protein-coupled receptors for PRESTO-Tango: parallel receptorome expression and screening via transcriptional output, with transcriptional activation following arrestin translocation

Bryan Roth

Kroeze et al Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/119869/60e0068c-c854-47db-8574-63f4c0f5e061/addgene-plasmid-66387-sequence-119869.gbk

partial

LOCUS sequence-119869- 919 bp DNA linear UNA 25-AUG-2024 DEFINITION Expression of G protein-coupled receptors for PRESTO-Tango: parallel receptorome expression and screening via transcriptional output, with transcriptional activation following arrestin translocation. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 919) AUTHORS Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, Sciaky N, Roth BL TITLE PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome. JOURNAL Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20. PUBMED 25895059 REFERENCE 2 (bases 1 to 919) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nsmb.3014"; journalName: "Nat Struct Mol Biol"; date: "2015-05"; volume: "22"; issue: "5"; pages: "362-9" FEATURES Location/Qualifiers source 1..919 /mol_type="genomic DNA" /organism="unspecified" primer_bind 48..67 /label=T7 /note="T7 promoter, forward primer" promoter 48..66 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac 61 tatagggaga cccaagctgg ctagcgttta aacttaagct tggtaccgag ctcggatcca 121 ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc gccaccatga 181 agacgatcat cgccctgagc tacatcttct gcctggtatt cgccgactac aaggacgatg 241 atgacgccag catcgatatg gttgggcttc tgctgttttt ttttccggca atatttttgg 301 aggtgagtct gctgccccgg tcacctggaa ggaaagttct gctcgccgga gctagtagcc 361 agcgcagcgt ggcaagaatg gacggagacg ttatcatcgg cgcactgttc agtgtacacc 421 accaaccacc agctgaaaag gtgccagagc ggaaatgtgg cgagattaga gagcaatacg 481 gaattcagcg agttgaggcc atgttccata cactcgacaa aataaatgca gaccctgtcc 541 tgctgcctaa tatcactttg ggaagcgaga tcagagacag ctgctggcac agctcagttg 601 ccttggagca gtctatagag ttcatcaggg acagcctcat ttccatccgc gacgagaagg 661 acggaataaa ccgatgcctg ccagacgggc agtcactgcc acccggtaga actaagaaac 721 ccatagccgg ggttataggg ccaggatcct ctagcgtggc catccaggta cagaatctgc 781 tgcaactgtt tgacatcccc cagatagcct actccgccac ttctatagac ttgagcgaca 841 agaccctcta caagtacttc cttagggttg tgccttccga taccctccag gcacgcgcga 901 tgctggatat cgtgaaacg //

pcDNA3-HA-REDD1 1-225 (#431)

99927

mammalian expression

James Brugarolas

Vega-Rubin-de-Celis et al Biochemistry. 2010 Mar 23;49(11):2491-501. doi: 10.1021/bi902135e.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/198007/aa1327ed-e0d0-42ca-81f1-924551b6dab6/addgene-plasmid-99927-sequence-198007.gbk

full

LOCUS sequence-198007- 6131 bp DNA circular SYN 26-AUG-2024 DEFINITION mammalian expression. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6131) AUTHORS Vega-Rubin-de-Celis S, Abdallah Z, Kinch L, Grishin NV, Brugarolas J, Zhang X TITLE Structural analysis and functional implications of the negative mTORC1 regulator REDD1. JOURNAL Biochemistry. 2010 Mar 23;49(11):2491-501. doi: 10.1021/bi902135e. PUBMED 20166753 REFERENCE 2 (bases 1 to 6131) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: "Biochemistry"; date: "2010-03-23- 23"; volume: "49"; issue: "11"; pages: "2491-501" FEATURES Location/Qualifiers source 1..6131 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1360..1386 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(1450..1468) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1451..1468) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1488..1505) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1494..1718 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1764..2192 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1851..1870) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2061..2082 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2201..2221) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2206..2535 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2386..2521 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2448..2467 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2602..3396 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2656..2675) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3266..3285 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3570..3691 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3607..3626) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3661..3680 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3740..3756) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3740..3756) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3753..3775) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3764..3780 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3788..3818) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3833..3854 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3971..3988) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4142..4730) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4222..4241) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4901..5761) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5524..5543 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5762..5866) /gene="bla" /label=AmpR promoter primer_bind complement(5941..5960) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt 661 ggtaccgagc tcggatccgc caccatgcct agcctttggg accgcttctc gtcgtcgtcc 721 acctcctctt cgccctcgtc cttgccccga actcccaccc cagatcggcc gccgcgctca 781 gcctgggggt cggcgacccg ggaggagggg tttgaccgct ccacgagcct ggagagctcg 841 gactgcgagt ccctggacag cagcaacagt ggcttcgggc cggaggaaga cacggcttac 901 ctggatgggg tgtcgttgcc cgactttgag ctgctcagtg accctgagga tgaacacttg 961 tgtgccaacc tgatgcagct gctgcaggag agcctggccc aggcgcggct gggctctcga 1021 cgccctgcgc gcctgctgat gcctagccag ttggtaagcc aggtgggcaa agaactactg 1081 cgcctggcct acagcgagcc gtgcggcctg cggggggcgc tgctggacgt ctgcgtggag 1141 cagggcaaga gctgccacag cgtgggccag ctggcactcg accccagcct ggtgcccacc 1201 ttccagctga ccctcgtgct gcgcctggac tcacgactct ggcccaagat ccaggggctg 1261 tttagctccg ccaactctcc cttcctccct ggcttcagcc agtccctgac gctgagcact 1321 ggcttccgag tcatcaagaa gaagctgtac agctcggaat atccctatga cgtcccagat 1381 tacgcttgag aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga 1441 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc 1501 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1561 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1621 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 1681 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 1741 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 1801 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 1861 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 1921 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 1981 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 2041 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 2101 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 2161 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 2221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2281 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2341 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 2401 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 2461 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 2521 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 2581 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 2641 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 2701 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 2761 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 2821 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 2881 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 2941 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 3001 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 3061 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 3121 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 3181 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 3241 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 3301 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 3361 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 3421 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 3481 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 3541 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 3601 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 3661 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 3721 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 3781 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 3841 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 3901 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 3961 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4021 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4081 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4561 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 4621 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 4681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 4741 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 4801 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 4861 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 4921 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 4981 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5041 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5101 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5161 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 5221 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 5281 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 5341 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 5401 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 5461 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 5521 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 5581 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 5641 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 5701 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 5761 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 5821 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 5881 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag 5941 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga 6001 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa 6061 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag 6121 atatacgcgt t //

pMM766

133603

Part Plasmid for dCas9 NLS Stop, Part 3b(coding sequence)

Megan McClean

Geller et al Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/267108/70eedd74-33be-46d2-8ac7-cdfdf9a50069/addgene-plasmid-133603-sequence-267108.gbk

full

LOCUS sequence-267108- 5817 bp DNA circular SYN 26-AUG-2024 DEFINITION Part Plasmid for dCas9 NLS Stop, Part 3b(coding sequence). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5817) AUTHORS Geller SH, Antwi EB, Di Ventura B, McClean MN TITLE Optogenetic Repressors of Gene Expression in Yeasts Using Light-Controlled Nuclear Localization. JOURNAL Cell Mol Bioeng. 2019 Sep 24;12(5):511-528. doi: 10.1007/s12195-019-00598-9. eCollection 2019 Oct. PUBMED 31719930 REFERENCE 2 (bases 1 to 5817) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1007/s12195-019-00598-9"; journalName: "Cell Mol Bioeng"; date: "2019-09-24- 24"; volume: "12"; issue: "5"; pages: "511-528" FEATURES Location/Qualifiers source 1..5817 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 4168..4188 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" terminator complement(4226..4260) /label=lambda t0 terminator /note="minimal transcription terminator from phage lambda (Scholtissek and Grosse, 1987)" CDS complement(4331..4990) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 4905..4924 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" rep_origin 5172..5760 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5661..5680 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atgttctttc ctgcgttatc ccctgattct gtggataacc gtagtcggtc tcattctgac 61 aagaagtatt ctatcggact ggccatcggg actaatagcg tcgggtgggc cgtgatcact 121 gacgagtaca aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc 181 atcaagaaaa atctgatcgg agctctcctc tttgattcag gggaaaccgc tgaagcaacc 241 cgcctcaagc ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt 301 caagagatat tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa 361 gaatcattcc tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc 421 gtcgacgagg tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg 481 gtggactcaa ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc 541 aagttcagag gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat 601 aaactgttca tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat 661 gcaagcggag tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt 721 gagaatctta tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt 781 gctctttcac ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc 841 aagctgcaac tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc 901 ggtgaccaat acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg 961 tctgacattc tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt 1021 aagcggtatg atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag 1081 cttccggaga agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac 1141 atcgacggag gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag 1201 atggacggca ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag 1261 cgcactttcg acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc 1321 cttcggcgcc aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa 1381 attctcactt tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt 1441 gcttggatga ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg 1501 gacaagggag ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc 1561 cccaatgaga aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac 1621 gagctgacca aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc 1681 gaacaaaaga aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag 1741 cagctgaagg aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg 1801 gtcgaagatc ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag 1861 gacaaggatt ttctggacaa tgaggagaaa gaggacatcc ttgaggacat tgtcctgact 1921 ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg 1981 ttcgacgata aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt 2041 agccgcaaac tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc 2101 cttaagagcg acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg 2161 acctttaagg aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag 2221 catatcgcaa atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa 2281 gtcgtggacg agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag 2341 atggccaggg aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa 2401 agaattgagg aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa 2461 aacacccagc ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg 2521 tatgtggacc aagagcttga tatcaacagg ctctcagact acgacgtgga cgccatcgtc 2581 cctcagagct tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag 2641 aaccggggaa agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat 2701 tggcgccagc ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag 2761 gctgagaggg gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg 2821 gagactcggc agattactaa acatgtagcc caaatccttg actcacgcat gaataccaag 2881 tacgacgaaa acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg 2941 gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat 3001 catgctcatg acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca 3061 aagctggaaa gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt 3121 gccaaatctg agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc 3181 atgaatttct tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc 3241 atcgagacta atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc 3301 gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc 3361 ggaggctttt ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg 3421 aagaaggatt gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct 3481 gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa 3541 ctgctgggta tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc 3601 gaagccaaag gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc 3661 ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag 3721 ggtaatgagc tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac 3781 gagaagctga aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat 3841 aagcattatc tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc 3901 gctgatgcca acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt 3961 cgggagcagg ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc 4021 gcattcaagt acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg 4081 ctggacgcca ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc 4141 tcacagctcg gaggggatga gggagctccc aagaaaaagc gcaaggtagg tagttccgga 4201 tcctagggat cctgagacca gaccaataaa aaacgcccgg cggcaaccga gcgttctgaa 4261 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct 4321 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca 4381 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca 4441 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt 4501 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgaaacga 4561 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca 4621 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg 4681 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata 4741 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg 4801 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa 4861 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg 4921 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt 4981 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg 5041 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaatcatg 5101 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 5161 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 5221 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 5281 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 5341 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 5401 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 5461 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 5521 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 5581 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 5641 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 5701 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 5761 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcac //

pRSET a MatryoshCaMP6s

100019

Bacterial expression of fluorescent reporter for calcium signaling, based off of GCaMP6s. Contains a stable reference Large Stokes Shift (LSS) mOrange nested within the reporting cpEGFP.

Wolf Frommer

Ast et al Nat Commun. 2017 Sep 5;8(1):431. doi: 10.1038/s41467-017-00400-2.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/194295/a2df309e-9e67-433a-97b0-b86413f68407/addgene-plasmid-100019-sequence-194295.gbk

full

LOCUS sequence-194295- 4932 bp DNA circular SYN 26-AUG-2024 DEFINITION Bacterial expression of fluorescent reporter for calcium signaling, based off of GCaMP6s. Contains a stable reference Large Stokes Shift (LSS) mOrange nested within the reporting cpEGFP.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4932) AUTHORS Ast C, Foret J, Oltrogge LM, De Michele R, Kleist TJ, Ho CH, Frommer WB TITLE Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins. JOURNAL Nat Commun. 2017 Sep 5;8(1):431. doi: 10.1038/s41467-017-00400-2. PUBMED 28874729 REFERENCE 2 (bases 1 to 4932) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun."; date: "2017-09-5"; volume: "8(1):431. doi"; pages: " 10.1038/s41467-017-00400-2" FEATURES Location/Qualifiers source 1..4932 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 51..73 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 93..110 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 114..146 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 150..173 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" regulatory 180..189 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 201..257 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 470..491 /label=EGFP-C /note="EGFP, forward primer" CDS 543..1250 /codon_start=1 /product="monomeric orange fluorescent protein with a large Stokes shift (Shcherbakova et al., 2012)" /label=LSSmOrange /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind 956..975 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(1305..1326) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1566..1585) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind complement(2314..2332) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2328..2375 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2472..2927 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2559..2578) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2769..2790 /label=F1ori-F /note="F1 origin, forward primer" promoter 2953..3057 /gene="bla" /label=AmpR promoter CDS 3058..3918 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3276..3295) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4089..4677 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4578..4597 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4831..4848 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac 61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta 121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatctcg 181 ccaccatggt cgactcatca cgtcgtaagt ggaataagac aggtcacgca gtcagagcta 241 taggtcggct gagctcactc gagaacgtct atatcaaggc cgacaagcag aagaacggca 301 tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc 361 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 421 tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc 481 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta 541 ccatggtgag caagggcgag gagaataaca tggccatcat caaggagttc atgcgcttca 601 aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg 661 gccgccccta cgagggcttt cagaccgtta agctgaaggt gaccaagggt ggccccctgc 721 ccttcgcctg ggacatcttg tcccctcagt tcacctacgg ctccaaggcc tacgtgaagc 781 accccgccga catccccgac tacctcaagc tgtccttccc cgagggcttc aagtgggagc 841 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac tcaggactcc tccctgcagg 901 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg 961 taatgcagaa gaagaccatg ggcatggagg cctcctccga gcggatgtac cccgaggacg 1021 gcgccctgaa gggcgaggac aagctcaggc tgaagctgaa ggacggcggc cactacacct 1081 ccgaggtcaa gaccacctac aaggccaaga agcccgtgca gttgcccggc gcctacatcg 1141 tcgacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg 1201 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggagggagca 1261 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 1321 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg 1381 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 1441 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 1501 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct 1561 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 1621 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 1681 agctggagta caacctgccg gaccaactga ctgaagagca gatcgcagaa tttaaagagg 1741 ctttctccct atttgacaag gacggggatg ggacaataac aaccaaggag ctggggacgg 1801 tgatgcggtc tctggggcag aaccccacag aagcagagct gcaggacatg atcaatgaag 1861 tagatgccga cggtgacggc acaatcgact tccctgagtt cctgacaatg atggcaagaa 1921 aaatgaaata cagggacacg gaagaagaaa ttagagaagc gttcggtgtg tttgataagg 1981 atggcaatgg ctacatcagt gcagcagagc ttcgccacgt gatgacaaac cttggagaga 2041 agttaacaga tgaagaggtt gatgaaatga tcagggaagc agacatcgat ggggatggtc 2101 aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gtgagcggcc gcgactctag 2161 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 2221 ctccccctga acctgaaaca taaaatgaat gcaattcgaa gcttgatccg gctgctaaca 2281 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc 2341 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat 2401 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 2461 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 2521 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 2581 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 2641 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 2701 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 2761 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 2821 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 2881 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt 2941 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 3001 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 3061 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 3121 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 3181 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 3241 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 3301 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 3361 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 3421 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 3481 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 3541 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 3601 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 3661 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 3721 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 3781 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 3841 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 3901 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 3961 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 4021 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 4081 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 4141 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 4201 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 4261 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 4321 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 4381 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 4441 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 4501 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 4561 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 4621 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 4681 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 4741 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 4801 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 4861 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggatctcg 4921 atcccgcgaa at //

pEPYC1CB0608

196172

MoClo Level 1 luciferase reporter: MinSyn_311 reporter [MinSyn_308::TMV::LucF::flag::ocsT]

Nicola Patron

Cai et al ACS Synth Biol. 2023 Nov 17;12(11):3482-3486. doi: 10.1021/acssynbio.3c00094. Epub 2023 Oct 19.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/386918/2b10e3b0-8209-4dc9-a3f0-ce0dd4a5c3fa/addgene-plasmid-196172-sequence-386918.gbk

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LOCUS sequence-386918- 7006 bp DNA circular SYN 27-AUG-2024 DEFINITION MoClo Level 1 luciferase reporter: MinSyn_311 reporter [MinSyn_308::TMV::LucF::flag::ocsT]. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7006) AUTHORS Cai YM, Witham S, Patron NJ TITLE Tuning Plant Promoters Using a Simple Split Luciferase Method to Assess Transcription Factor-DNA Interactions. JOURNAL ACS Synth Biol. 2023 Nov 17;12(11):3482-3486. doi: 10.1021/acssynbio.3c00094. Epub 2023 Oct 19. PUBMED 37856867 REFERENCE 2 (bases 1 to 7006) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1021/acssynbio.3c00094"; journalName: "ACS Synth Biol"; date: "2023-11-17- 17"; volume: "12"; issue: "11"; pages: "3482-3486" FEATURES Location/Qualifiers source 1..7006 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 42..66 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS complement(222..1370) /codon_start=1 /product="trans-acting replication protein that binds to and activates oriV" /label=trfA /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" rep_origin complement(1706..2294) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1786..1805) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2465..3325) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3088..3107 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3326..3430) /gene="bla" /label=AmpR promoter rep_origin 3517..4148 /label=oriV /note="incP origin of replication" misc_feature 4240..4264 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" misc_feature 4488..4543 /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" CDS 4545..6194 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(4611..4629) /label=LucNrev /note="Firefly luciferase, reverse primer" CDS 6258..6281 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 6292..6999 /label=OCS terminator /note="octopine synthase terminator" ORIGIN 1 gcaattgtct tctgcacgaa gtggtttaaa ctatcagtgt ttgacaggat atattggcgg 61 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa 121 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc agatcaggcg 181 ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc 241 aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct 301 tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg 361 aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt 421 cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca 481 gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca 541 cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg 601 gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa 661 taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc 721 ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca 781 tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa 841 atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca 901 gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac 961 aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg 1021 gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata 1081 gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcaagacga 1141 cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg 1201 tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg gaaggtttcg 1261 cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc 1321 gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct 1381 tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt tgacccgcct 1441 ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc gtagaccgtc 1501 tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac cagcgacccc 1561 ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat gcggaagaag 1621 tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggatct gccaggaacc 1681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 1741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 1801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 1861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 1921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 1981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 2041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 2101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 2161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 2221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 2281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 2341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 2581 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa 2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 2761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 3061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 3361 aaggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 3421 gggttccgcg cacgaattgg ccagcgctgc catttttggg gtgaggccgt tcgcggccga 3481 ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag 3541 ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa 3601 caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa 3661 acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca 3721 ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc 3781 atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg 3841 tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg 3901 gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg 3961 gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt 4021 ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc acacggcttc gacggcgttt 4081 ctggcgcgtt tgcagggcca tagacggccg ccagcccagc ggcgagggca accagcccgg 4141 tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg cctgtatcga gtggtgattt 4201 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt 4261 aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg 4321 tggatgcagt gggccccact ctgtgaagac aatgccgaat tcggatccgg agggaggcgt 4381 gtcgttttag tgaggcttac gtctgggatt acgtctacta tataaggttt tgctattcat 4441 tgaaagcagt agtgactgat ttgtatataa tgtactgtat ttttacaaca attaccaaca 4501 acaacaaaca acaaacaaca ttacaattac tatttacaat tacaatggag gacgccaaaa 4561 acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac 4621 tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac 4681 atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag 4741 ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc 4801 ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga 4861 acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg 4921 tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca 4981 tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca 5041 cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct 5101 tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta 5161 aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta 5221 tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg 5281 gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt 5341 atagatttga agaagaactg tttctgagga gccttcagga ttacaagatt caaagtgcgc 5401 tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt 5461 tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag 5521 cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta 5581 catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg 5641 ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc 5701 aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg 5761 aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact 5821 gggacgagga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag 5881 gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg 5941 acgcaggtgt cgcaggtctg cccgacgatg acgccggtga acttcccgcc gccgttgttg 6001 ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag 6061 taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc 6121 ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg 6181 gaaagatcgc cgtgggttcg gattataagg accatgacgg agactataag gaccatgacc 6241 tcgacgctgc agcagcggat tataaggacg atgacgataa gtgagcttgt cctgctttaa 6301 tgagatatgc gagaagccta tgatcgcatg atatttgctt tcaattctgt tgtgcacgtt 6361 gtaaaaaacc tgagcatgtg tagctcagat ccttaccgcc ggtttcggtt cattctaatg 6421 aatatatcac ccgttactat cgtattttta tgaataatat tctccgttca atttactgat 6481 tgtaccctac tacttatatg tacaatatta aaatgaaaac aatatattgt gctgaatagg 6541 tttatagcga catctatgat agagcgccac aataacaaac aattgcgttt tattattaca 6601 aatccaattt taaaaaaagc ggcagaaccg gtcaaaccta aaagactgat tacataaatc 6661 ttattcaaat ttcaaaagtg ccccaggggc tagtatctac gacacaccga gcggcgaact 6721 aataacgctc actgaaggga actccggttc cccgccggcg cgcatgggtg agattccttg 6781 aagttgagta ttggccgtcc gctctaccga aagttacggg caccattcaa cccggtccag 6841 cacggcggcc gggtaaccga cttgctgccc cgagaattat gcagcatttt tttggtgtat 6901 gtgggcccca aatgaagtgc aggtcaaacc ttgacagtga cgacaaatcg ttgggcgggt 6961 ccagggcgaa ttttgcgaca acatgtcgag gctcagcagg accgct //

pORFE0002

112071

AtCAS9 nickase D10A module CDS1

Christian Chevalier

Chevalier Lab plasmids (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/217629/8a3c04e8-f14e-459a-a393-665b698cfbb6/addgene-plasmid-112071-sequence-217629.gbk

full

LOCUS sequence-217629- 6390 bp DNA circular SYN 26-AUG-2024 DEFINITION AtCAS9 nickase D10A module CDS1. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6390) TITLE Chevalier Lab plasmids REFERENCE 2 (bases 1 to 6390) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..6390 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 1..4104 /codon_start=1 /product="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9(D10A) /note="generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 4117..4137 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(4180..4198) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4266..4370 /gene="bla" /label=AmpR promoter CDS 4592..5383 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 5554..6142 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6043..6062 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6296..6313 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 atggataaga agtactctat cggactcgct atcggaacta actctgtggg atgggctgtg 61 atcaccgatg agtacaaggt gccatctaag aagttcaagg ttctcggaaa caccgatagg 121 cactctatca agaaaaacct tatcggtgct ctcctcttcg attctggtga aactgctgag 181 gctaccagac tcaagagaac cgctagaaga aggtacacca gaagaaagaa caggatctgc 241 tacctccaag agatcttctc taacgagatg gctaaagtgg atgattcatt cttccacagg 301 ctcgaagagt cattcctcgt ggaagaagat aagaagcacg agaggcaccc tatcttcgga 361 aacatcgttg atgaggtggc ataccacgag aagtacccta ctatctacca cctcagaaag 421 aagctcgttg attctactga taaggctgat ctcaggctca tctacctcgc tctcgctcac 481 atgatcaagt tcagaggaca cttcctcatc gagggtgatc tcaaccctga taactctgat 541 gtggataagt tgttcatcca gctcgtgcag acctacaacc agcttttcga agagaaccct 601 atcaacgctt caggtgtgga tgctaaggct atcctctctg ctaggctctc taagtcaaga 661 aggcttgaga acctcattgc tcagctccct ggtgagaaga agaacggact tttcggaaac 721 ttgatcgctc tctctctcgg actcacccct aacttcaagt ctaacttcga tctcgctgag 781 gatgcaaagc tccagctctc aaaggatacc tacgatgatg atctcgataa cctcctcgct 841 cagatcggag atcagtacgc tgatttgttc ctcgctgcta agaacctctc tgatgctatc 901 ctcctcagtg atatcctcag agtgaacacc gagatcacca aggctccact ctcagcttct 961 atgatcaaga gatacgatga gcaccaccag gatctcacac ttctcaaggc tcttgttaga 1021 cagcagctcc cagagaagta caaagagatt ttcttcgatc agtctaagaa cggatacgct 1081 ggttacatcg atggtggtgc atctcaagaa gagttctaca agttcatcaa gcctatcctc 1141 gagaagatgg atggaaccga ggaactcctc gtgaagctca atagagagga tcttctcaga 1201 aagcagagga ccttcgataa cggatctatc cctcatcaga tccacctcgg agagttgcac 1261 gctatcctta gaaggcaaga ggatttctac ccattcctca aggataacag ggaaaagatt 1321 gagaagattc tcaccttcag aatcccttac tacgtgggac ctctcgctag aggaaactca 1381 agattcgctt ggatgaccag aaagtctgag gaaaccatca ccccttggaa cttcgaagag 1441 gtggtggata agggtgctag tgctcagtct ttcatcgaga ggatgaccaa cttcgataag 1501 aaccttccaa acgagaaggt gctccctaag cactctttgc tctacgagta cttcaccgtg 1561 tacaacgagt tgaccaaggt taagtacgtg accgagggaa tgaggaagcc tgcttttttg 1621 tcaggtgagc aaaagaaggc tatcgttgat ctcttgttca agaccaacag aaaggtgacc 1681 gtgaagcagc tcaaagagga ttacttcaag aaaatcgagt gcttcgattc agttgagatt 1741 tctggtgttg aggataggtt caacgcatct ctcggaacct accacgatct cctcaagatc 1801 attaaggata aggatttctt ggataacgag gaaaacgagg atatcttgga ggatatcgtt 1861 cttaccctca ccctctttga agatagagag atgattgaag aaaggctcaa gacctacgct 1921 catctcttcg atgataaggt gatgaagcag ttgaagagaa gaagatacac tggttgggga 1981 aggctctcaa gaaagctcat taacggaatc agggataagc agtctggaaa gacaatcctt 2041 gatttcctca agtctgatgg attcgctaac agaaacttca tgcagctcat ccacgatgat 2101 tctctcacct ttaaagagga tatccagaag gctcaggttt caggacaggg tgatagtctc 2161 catgagcata tcgctaacct cgctggatct cctgcaatca agaagggaat cctccagact 2221 gtgaaggttg tggatgagtt ggtgaaggtg atgggaaggc ataagcctga gaacatcgtg 2281 atcgaaatgg ctagagagaa ccagaccact cagaagggac agaagaactc tagggaaagg 2341 atgaagagga tcgaggaagg tatcaaagag cttggatctc agatcctcaa agagcaccct 2401 gttgagaaca ctcagctcca gaatgagaag ctctacctct actacctcca gaacggaagg 2461 gatatgtatg tggatcaaga gttggatatc aacaggctct ctgattacga tgttgatcat 2521 atcgtgccac agtcattctt gaaggatgat tctatcgata acaaggtgct caccaggtct 2581 gataagaaca ggggtaagag tgataacgtg ccaagtgaag aggttgtgaa gaaaatgaag 2641 aactattgga ggcagctcct caacgctaag ctcatcactc agagaaagtt cgataacttg 2701 actaaggctg agaggggagg actctctgaa ttggataagg caggattcat caagaggcag 2761 cttgtggaaa ccaggcagat cactaagcac gttgcacaga tcctcgattc taggatgaac 2821 accaagtacg atgagaacga taagttgatc agggaagtga aggttatcac cctcaagtca 2881 aagctcgtgt ctgatttcag aaaggatttc caattctaca aggtgaggga aatcaacaac 2941 taccaccacg ctcacgatgc ttaccttaac gctgttgttg gaaccgctct catcaagaag 3001 tatcctaagc tcgagtcaga gttcgtgtac ggtgattaca aggtgtacga tgtgaggaag 3061 atgatcgcta agtctgagca agagatcgga aaggctaccg ctaagtattt cttctactct 3121 aacatcatga atttcttcaa gaccgagatt accctcgcta acggtgagat cagaaagagg 3181 ccactcatcg agacaaacgg tgaaacaggt gagatcgtgt gggataaggg aagggatttc 3241 gctaccgtta gaaaggtgct ctctatgcca caggtgaaca tcgttaagaa aaccgaggtg 3301 cagaccggtg gattctctaa agagtctatc ctccctaaga ggaactctga taagctcatt 3361 gctaggaaga aggattggga ccctaagaaa tacggtggtt tcgattctcc taccgtggct 3421 tactctgttc tcgttgtggc taaggttgag aagggaaaga gtaagaagct caagtctgtt 3481 aaggaacttc tcggaatcac tatcatggaa aggtcatctt tcgagaagaa cccaatcgat 3541 ttcctcgagg ctaagggata caaagaggtt aagaaggatc tcatcatcaa gctcccaaag 3601 tactcactct tcgaactcga gaacggtaga aagaggatgc tcgcttctgc tggtgagctt 3661 caaaagggaa acgagcttgc tctcccatct aagtacgtta actttcttta cctcgcttct 3721 cactacgaga agttgaaggg atctccagaa gataacgagc agaagcaact tttcgttgag 3781 cagcacaagc actacttgga tgagatcatc gagcagatct ctgagttctc taaaagggtg 3841 atcctcgctg atgcaaacct cgataaggtg ttgtctgctt acaacaagca cagagataag 3901 cctatcaggg aacaggcaga gaacatcatc catctcttca cccttaccaa cctcggtgct 3961 cctgctgctt tcaagtactt cgatacaacc atcgatagga agagatacac ctctaccaaa 4021 gaagtgctcg atgctaccct catccatcag tctatcactg gactctacga gactaggatc 4081 gatctctcac agctcggtgg tgattcaagg gctgatccta agaagaagag gaaggtttga 4141 gctttgagac cacgaagtgg ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt 4201 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa 4261 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 4321 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgcgctcac 4381 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca 4441 tggcttgtta tgactgtttt tttttggggt acagtctatg cctcgggcat ccaagcagca 4501 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca 4561 gggcagtcgc cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc 4621 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc 4681 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagcg atattgattt 4741 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct 4801 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat 4861 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg 4921 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga 4981 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc 5041 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt 5101 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt 5161 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga 5221 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc 5281 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt 5341 ccattacgta aaaggcgaga tcaccaaggt agtcggcaaa taactgtcag accaagttta 5401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 5461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 5521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 5581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 5641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 5701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 5761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 5821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 5881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 5941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 6001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 6061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 6121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 6181 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 6241 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 6301 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 6361 gccgattcat taatcactct gtggtctcaa //

pcDNA3.2/V5 mmu-mir-133a-1

26326

Low

David Bartel

Chiang et al Genes Dev. 2010 May 15. 24(10):992-1009.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/32478/35cc0c29-13e6-4e4b-9c90-124b89d78631/addgene-plasmid-26326-sequence-32478.gbk

partial

LOCUS sequence-32478-e 954 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 954) AUTHORS Chiang HR, Schoenfeld LW, Ruby JG, Auyeung VC, Spies N, Baek D, Johnston WK, Russ C, Luo S, Babiarz JE, Blelloch R, Schroth GP, Nusbaum C, Bartel DP TITLE Mammalian microRNAs: experimental evaluation of novel and previously annotated genes. JOURNAL Genes Dev. 2010 May 15. 24(10):992-1009. PUBMED 20413612 REFERENCE 2 (bases 1 to 954) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genes Dev. 2010 May 15. 24(10):992-1009." FEATURES Location/Qualifiers source 1..954 /mol_type="genomic DNA" /organism="unspecified" primer_bind 40..59 /label=T7 /note="T7 promoter, forward primer" promoter 40..58 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 88..112 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(431..455) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 481..522 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(562..581) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 606..654 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 ggctactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga 61 gacccaagct ggctagttaa gctatcaaca agtttgtaca aaaaagcagg ctgcctaaac 121 acgtgaattt tctgtttaac aactggtaca ctagtgtggg aacctctaat acctgtcatg 181 ctatatttct acaaaagagc atttaacctg tttggatcct acaccagcag cggcaatgct 241 ttgctaaagc tggtaaaatg gaaccaaatc gcctcttcaa tggatttggt ccccttcaac 301 cagctgtagc tatgcattga ttacacatgg acagccagtg ttttatgagc atgaattaca 361 tctcaactta ctcaattcat acaaactgta actaaaagca aatacctagc aactgtcaac 421 cttatttgga acccagcttt cttgtacaaa gtggttgatc tagagggccc gcggttcgaa 481 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg ttagtaatga 541 gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc 601 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac 661 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc 721 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag 781 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct 841 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggg tgtggtggtt 901 acccgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgct //

pBBK72 pHomL-RPL18Bp-Scarlet(rfp)-Link-SCS2tmh-SSA1t-HomR (AmpR)

179056

Vector containing a yeast integration cassette for RPL18B-driven expression of Scarlet(rfp)-Link-SCS2tmh, a marker for the endoplasmic reticulum

Vincent Martin

Bean et al G3 (Bethesda). 2022 Jun 16. pii: 6609175. doi: 10.1093/g3journal/jkac154.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/360861/3609e4b1-a6a2-40ba-9345-1bf8522c1c34/addgene-plasmid-179056-sequence-360861.gbk

full

LOCUS sequence-360861- 5724 bp DNA circular SYN 26-AUG-2024 DEFINITION Vector containing a yeast integration cassette for RPL18B-driven expression of Scarlet(rfp)-Link-SCS2tmh, a marker for the endoplasmic reticulum. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5724) AUTHORS Bean BDM, Whiteway M, Martin VJJ TITLE The MyLo CRISPR-Cas9 Toolkit: A Markerless Yeast Localization and Overexpression CRISPR-Cas9 Toolkit. JOURNAL G3 (Bethesda). 2022 Jun 16. pii: 6609175. doi: 10.1093/g3journal/jkac154. PUBMED 35708612 REFERENCE 2 (bases 1 to 5724) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "G3 (Bethesda). 2022 Jun 16. pii: 6609175. doi: 10.1093/g3journal/jkac154." FEATURES Location/Qualifiers source 1..5724 /mol_type="other DNA" /organism="synthetic DNA construct" CDS complement(1..861) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 624..643 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(862..966) /gene="bla" /label=AmpR promoter rep_origin 1069..1657 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1558..1577 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 121 cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa 181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 961 tccgcgcaca tttccccgaa aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt 1021 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 1081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 1141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 1201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 1261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 1321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 1381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 1441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 1501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 1561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 1621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 1681 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 1741 ttctgtggat aaccgtgcgg ccgccaatgt tatctattgg cagatgttca tacggggctt 1801 tcaaatattg atgaaattat gtgatgttta gaagaagatt cgaactgttt tcagtagatt 1861 tggtaactgt gcaaccataa ctcatgcccc ctatttatcg gcattgagat tgtaaaggaa 1921 gtaaaacgca tttttgcaga tctgttctct tatgtatttt taatcgtcct tgtatggaag 1981 tatcaaaggg gacgttcttc acctccttgg aaaacgcaaa ctttcttttt gcctatcaca 2041 gtatcttatc gagctaacta ttttcgacac acatgaaaaa gcagaaatat taacgaaaaa 2101 gaaaagaaag accatgtcat gtacgggcaa tcagaatatg caagatgagc gggaacgggc 2161 gaaatgtgca cgcgcacaac ttacgaaacg cggatgagtc actgacagcc accgcagagg 2221 ttctgactcc tactgagctc tattggaggt ggcagaaccg gtaatcccac agtgagcaac 2281 ttgacaaacc aacagatata ggctgtgtct taaaatatac catccaatac cttgatgaac 2341 ttttcaaaga tgactaaaat aagtgaaatt tcaacattaa cttcgagctg taaacacagt 2401 ggtttctttg cataaacacc atcagcctca agtcgtcaag taaagatttc gtgttcatgc 2461 agatagataa caatctatat gttgataatt agcgttgcct catcaatgcg tacacgaaga 2521 atatactaaa aaatgagcag gcaagataaa cgaaggcaaa gatgacagag cagaaagccc 2581 tagtaaagcg tattacaaat gaaaccaaga ttcagattgc gatccctgaa ttcgcatcta 2641 gatggtagag ccacaaacag ccggtacaag caacgatctc caggaccatc tgaatcatgc 2701 gcggatgaca cgaactcacg acggcgatca cagacattaa cccacagtac agacactgcg 2761 acaacgtggc aattcgtcgc aataccgtct cactgaactg gccgataatt gcagacgaac 2821 gaagaggatg tccaatattt tttttaagga ataaggatac ttcaagacta gattcccccc 2881 tgcattccca tcagaaccgt aaaccttggc gctttccttg ggaagtattc aagaagtgcc 2941 ttgtccggtt tctgtggctc acaaaccagc gcgcccgata tggctttctt ttcacttatg 3001 aatgtaccag tacgggacaa ttagaacgct cctgtaacaa tctctttgca aatgtggggt 3061 tacattctaa ccatgtcaca ctgctgacga aattcaaagt aaaaaaaaat gggaccacgt 3121 cttgagaacg atagattttc tttattttac attgaacagt cgttgtctca gcgcgcttta 3181 tgttttcatt catacttcat attataaaat aacaaaagaa gaatttcata ttcacgccca 3241 agaaatcagg ctgctttcca aatgcaattg acacttcatt agccatcaca caaaactctt 3301 tcttgctgga gcttctttta aaaaagacct cagtacacca aacacgttac ccgacctcgt 3361 tattttacga caactatgat aaaattctga agaaaaaata aaaaaatttt catacttctt 3421 gcttttattt aaaccattga atgatttctt ttgaacaaaa ctacctgttt caccaaagga 3481 aatagaaaga aaaaatcaat tagaagaaaa caaaaaacaa aagatctatg gttagtaaag 3541 gtgaagctgt tataaaagaa ttcatgaggt ttaaagttca tatggaaggt tcaatgaatg 3601 gtcatgaatt tgaaattgaa ggtgaaggtg aaggtagacc atatgaaggt acacaaactg 3661 ctaaattgaa agttactaaa ggtggtccat taccattttc ttgggatatt ttgtctccac 3721 aattcatgta tggttctaga gcttttatta agcatccagc tgatattcca gattattata 3781 aacaatcttt tcctgaaggt tttaaatggg aaagagttat gaatttcgaa gatggtggtg 3841 ctgttactgt tactcaagat acttctttgg aagatggtac tttaatctat aaagttaaat 3901 tgagaggtac taattttcca ccagatggtc cagttatgca aaagaaaact atgggttggg 3961 aagcatctac tgaaagattg tatccagaag atggtgtttt gaaaggagac attaagatgg 4021 ctttgagatt gaaagatggt ggtcggtacc tggccgactt caagaccacc tataaagcta 4081 aaaaaccagt tcaaatgcca ggtgcatata atgttgatag aaagttagat ataacgtcgc 4141 ataacgagga ctatactgta gttgaacaat atgaacgtag tgaaggtaga catagtaccg 4201 gaggaatgga tgaattgtat aaatctggtc cttctggccc aactggttct aatgaatcat 4261 ccagcatggg tatattcata ttggttgcac tccttatctt ggttttagga tggttctaca 4321 gatagcgaga tcctaactcg aggccaattg gtgcggcaat tgataataac gaaaatgtct 4381 tttaatgatc tgggtataat gaggaatttt ccgaacgttt ttactttata tatatatata 4441 catgtaacat atattctata cgctatagag aaaggaaatt tttcaattaa aaaaaaaata 4501 gagaaagagt ttcacttctt gattatcgct aacactaatg gttgaagtac tgctacttta 4561 attttatgct gccaatgaga cgacggggtc atcacggctc atcatgcgcc aaacaaatgt 4621 gtgcaataca cgctcggatg actgcatgat gaccgcactg actggggaca gcagatccac 4681 ctaagcctgt gagagaagca gacacccgac agatcaaggc agttaactag tgcactgcag 4741 tacaaatcgt ccccaacaaa agtgggctct caaaattcat cacatttaaa tgcatatagg 4801 aagagcaaca gttggtttgc atctgatgtt ccttaaagat ttcgacataa tgtgcgaagt 4861 agatcccttc agcactgtac ttgtttttgc gactagattg taaatcattc tttatttaat 4921 ctctttcttt aactactgct taaagtataa tttggtccgt agtttaataa ctatactaag 4981 cgaacgctct ctcgaaattt ttcttaacgc gtccttgtac tgcgtctaac gcttttgcca 5041 cttggatttc tattatagga aatagtctca cttactgggc gacgaatttt cgcgttttga 5101 tgaagcacag gaagaatttc tttatgctca gaagtttgac agcaagcaag ttcatcattc 5161 gaactagcct tattgtttta gttcagtgac agcgaactgc cgtactcgat gctttatttc 5221 tcacggtaga gcggaagaac agatagatcc ctacatattg ctaccacttc tattacacaa 5281 tagtttcaat agcttgcagc gtagctaaac tctaaaattt atctaaatca ctcatataaa 5341 ccgaaccctt ccccttccgc ttatagtaca gctgatgctc cttcactatt ttaacatgtg 5401 gaattcttga aagaatgaaa tcgccatgcc aagccatcac acggtctttt atgcaattga 5461 ttgaccgcct gcaacacata ggcagtaaaa tttttacaag cagtagcaga acaggccaca 5521 caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat gatacatgct 5581 ctggccaagc attccggctg gtcgctaatc gttgagtgcc gagcggccgc gattatcaaa 5641 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5701 atatgagtaa acttggtctg acag //

VDAC2_R (OZ524)

27199

Keith Joung

Maeder et al Mol Cell. 2008 Jul 25. 31(2):294-301.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/33611/d937a7cd-4901-4099-af0f-11f8e35eb2c9/addgene-plasmid-27199-sequence-33611.gbk

partial

LOCUS sequence-33611-e 872 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 872) AUTHORS Maeder ML, Thibodeau-Beganny S, Osiak A, Wright DA, Anthony RM, Eichtinger M, Jiang T, Foley JE, Winfrey RJ, Townsend JA, Unger-Wallace E, Sander JD, Muller-Lerch F, Fu F, Pearlberg J, Goebel C, Dassie JP, Pruett-Miller SM, Porteus MH, Sgroi DC, Iafrate AJ, Dobbs D, McCray PB, Cathomen T, Voytas DF, Joung JK TITLE Rapid ""open-source"" engineering of customized zinc-finger nucleases for highly efficient gene modification. JOURNAL Mol Cell. 2008 Jul 25. 31(2):294-301. PUBMED 18657511 REFERENCE 2 (bases 1 to 872) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 2008 Jul 25. 31(2):294-301." FEATURES Location/Qualifiers source 1..872 /mol_type="genomic DNA" /organism="unspecified" CDS 45..68 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" rep_origin 470..850 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(557..576) /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 attgtgcaaa ggccctgaga aaaatccaag ctttggcggc cgctgactac aaagacgatg 61 atgacaaatc tagacccggg gagcgcccct tccagtgtcg catttgcatg cggaactttt 121 cgaccgcggc ggtgcttacc cggcataccc gtactcatac cggtgaaaaa ccgtttcagt 181 gtcggatctg tatgcgaaat ttctccctcc cccaccactt gcagcggcat ctacgtacgc 241 acaccggcga gaagccattc caatgccgaa tatgcatgcg caacttcagt ctcaagcacg 301 acctgcggag gcacctaaaa acccacctga ggggatcctc tgcacagtag taactagaga 361 ctagaaaaag gccgacaagt cccgctccgc tgaagatcct ggcgtaatag cgaagaggcc 421 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt 481 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 541 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 601 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 661 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 721 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaataatgg actcttgttc 781 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 841 ccgatttcgg cctattgtaa atatatacaa ta //

pT7-EGFP-C1-HseIF4G1-L640AV646A_AD

148520

Mammalian Expression of HseIF4G1-L640AV646A

Elisa Izaurralde

Gruner et al Mol Cell. 2016 Nov 3;64(3):467-479. doi: 10.1016/j.molcel.2016.09.020. Epub 2016 Oct 20.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/290968/288f2682-323d-4eb5-a530-a3477869e40d/addgene-plasmid-148520-sequence-290968.gbk

full

LOCUS sequence-290968- 9603 bp DNA circular SYN 26-AUG-2024 DEFINITION Mammalian Expression of HseIF4G1-L640AV646A. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9603) AUTHORS Gruner S, Peter D, Weber R, Wohlbold L, Chung MY, Weichenrieder O, Valkov E, Igreja C, Izaurralde E TITLE The Structures of eIF4E-eIF4G Complexes Reveal an Extended Interface to Regulate Translation Initiation. JOURNAL Mol Cell. 2016 Nov 3;64(3):467-479. doi: 10.1016/j.molcel.2016.09.020. Epub 2016 Oct 20. PUBMED 27773676 REFERENCE 2 (bases 1 to 9603) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.molcel.2016.09.020"; journalName: "Mol Cell"; date: "2016-11-3- 3"; volume: "64"; issue: "3"; pages: "467-479" FEATURES Location/Qualifiers source 1..9603 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 613..632 /label=T7 /note="T7 promoter, forward primer" promoter 613..631 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 659..668 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 665..1381 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 665..1381 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(710..731) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(971..990) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1318..1339 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 6391..6512 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6428..6447) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6482..6501 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(6519..6974) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6656..6677) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6868..6887 /label=F1ori-R /note="F1 origin, reverse primer" promoter 7001..7105 /gene="bla" /label=AmpR promoter promoter 7107..7464 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 7107..7298 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(7130..7150) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 7315..7450 /label=SV40 ori /note="SV40 origin of replication" CDS 7499..8293 /codon_start=1 /product="aminoglycoside phosphotransferase" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS 7499..8293 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7553..7572) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 8163..8182 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(8481..8500) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 8525..8572 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 8901..9489 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9390..9409 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg 601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc 661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct 721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac 781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc 841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat 901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat 961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac 1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg 1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa 1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct 1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa 1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat 1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa 1381 gtccggactc agatctcgag ctatgaacaa agctccacag tccacaggcc ccccacccgc 1441 cccatccccc ggactcccac agccagcgtt tcccccgggg cagacagcgc cggtggtgtt 1501 cagtacgcca caagcgacac aaatgaacac gccttctcag ccccgccagg gaggattcag 1561 gtctctgcag cacttctacc ctagccgggc ccagcccccg agcagtgcag cctcccgagt 1621 gcagagtgca gcccctgccc gccctggccc agctgcccat gtctaccctg ctggatccca 1681 agtaatgatg atcccttccc agatctccta cccagcctcc cagggggcct actacatccc 1741 tggacagggg cgttccacat acgttgtccc gacacagcag taccctgtgc agccaggagc 1801 cccaggcttc tatccaggtg caagccctac agaatttggg acctacgctg gcgcctacta 1861 tccagcccaa ggggtgcagc agtttcccac tggcgtggcc cccgccccag ttttgatgaa 1921 ccagccaccc cagattgctc ccaagaggga gcgtaagacg atccgaattc gagatccaaa 1981 ccaaggagga aaggatatca cagaggagat catgtctggg gcccgcactg cctccacacc 2041 cacccctccc cagacgggag gcggtctgga gcctcaagct aatggggaga cgccccaggt 2101 tgctgtcatt gtccggccag atgaccggtc acagggagca atcattgctg accggccagg 2161 gctgcctggc ccagagcata gcccttcaga atcccagcct tcgtcgcctt ctccgacccc 2221 atcaccatcc ccagtcttgg aaccggggtc tgagcctaat ctcgcagtcc tctctattcc 2281 tggggacact atgacaacta tacaaatgtc tgtagaagaa tcaaccccca tctcccgtga 2341 aactggggag ccatatcgcc tctctccaga acccactcct ctcgccgaac ccatactgga 2401 agtagaagtg acacttagca aaccggttcc agaatctgag ttttcttcca gtcctctcca 2461 ggctcccacc cctttggcat ctcacacagt ggaaattcat gagcctaatg gcatggtccc 2521 atctgaagat ctggaaccag aggtggagtc aagcccagag cttgctcctc ccccagcttg 2581 cccctccgaa tcccctgtgc ccattgctcc aactgcccaa cctgaggaac tgctcaacgg 2641 agccccctcg ccaccagctg tggacttaag cccagtcagt gagccagagg agcaggccaa 2701 ggaggtgaca gcatcagtgg cgccccccac catcccctct gctactccag ctacggctcc 2761 ttcagctact tccccagctc aggaggagga aatggaagaa gaagaagaag aggaagaagg 2821 agaagcagga gaagcaggag aagctgagag tgagaaagga ggagaggaac tgctcccccc 2881 agagagtacc cctattccag ccaacttgtc tcagaatttg gaggcagcag cagccactca 2941 agtggcagta tctgtgccaa agaggagacg gaaaattaag gagctaaata agaaggaggc 3001 tgttggagac cttctggatg ccttcaagga ggcgaacccg gcagtaccag aggtggaaaa 3061 tcagcctcct gcaggcagca atccaggccc agagtctgag ggcagtggtg tgcccccacg 3121 tcctgaggaa gcagatgaga cctgggactc aaaggaagac aaaattcaca atgctgagaa 3181 catccagccc ggggaacaga agtatgaata taagtcagat cagtggaagc ctctaaacct 3241 agaggagaaa aaacgttacg accgtgagtt cctgcttggt tttcagttca tctttgccag 3301 tatgcagaag ccagagggag ccccacatat cagtgacgcg gtgctggaca aggccaataa 3361 aacaccactg cggccactgg atcccactag actacaaggc ataaattgtg gcccagactt 3421 cactccatcc tttgccaacc ttggccggac aacccttagc acccgtgggc ccccaagggg 3481 tgggccaggt ggggagctgc cccgtgggcc ggctggcctg ggaccccggc gctctcagca 3541 gggaccccga aaagaaccac gcaagatcat tgccacagtg ttaatgaccg aagatataaa 3601 actgaacaaa gcagagaaag cctggaaacc cagcagcaag cggacggcgg ctgataagga 3661 tcgaggggaa gaagatgctg atggcagcaa aacccaggac ctattccgca gggtgcgctc 3721 catcctgaat aaactgacac cccagatgtt ccagcagctg atgaagcaag tgacgcagct 3781 ggccatcgac accgaggaac gcctcaaagg ggtcattgac ctcatttttg agaaggccat 3841 ttcagagccc aacttctctg tggcctatgc caacatgtgc cgctgcctca tggcgctgaa 3901 agtgcccact acggaaaagc caacagtgac tgtgaacttc cgaaagctgt tgttgaatcg 3961 atgtcagaag gagtttgaga aagacaaaga tgatgatgag gtttttgaga agaagcaaaa 4021 agagatggat gaagctgcta cggcagagga acgaggacgc ctgaaggaag agctggaaga 4081 ggctcgggac atagcccggc ggcgctcttt agggaatatc aagtttattg gagagttgtt 4141 caaactgaag atgttaacag aggcaataat gcatgactgt gtggtcaaac tgcttaagaa 4201 ccatgatgaa gagtcccttg agtgcctttg tcgtctgctc accaccattg gcaaagacct 4261 ggactttgaa aaagccaagc cccgaatgga tcagtatttc aaccagatgg aaaaaatcat 4321 taaagaaaag aagacgtcat cccgcatccg ctttatgctg caggacgtgc tggatctgcg 4381 agggagcaat tgggtgccac gccgagggga tcagggtccc aagaccattg accagatcca 4441 taaggaggct gagatggaag aacatcgaga gcacatcaaa gtgcagcagc tcatggccaa 4501 gggcagtgac aagcgtcggg gcggtcctcc aggccctccc atcagccgtg gacttcccct 4561 tgtggatgat ggtggctgga acacagttcc catcagcaaa ggtagccgcc ccattgacac 4621 ctcacgactc accaagatca ccaagcctgg ctccatcgat tctaacaacc agctctttgc 4681 acctggaggg cgactgagct ggggcaaggg cagcagcgga ggctcaggag ccaagccctc 4741 agacgcagca tcagaagctg ctcgcccagc tactagtact ttgaatcgct tctcagccct 4801 tcaacaagcg gtacccacag aaagcacaga taatagacgt gtggtgcaga ggagtagctt 4861 gagccgagaa cgaggcgaga aagctggaga ccgaggagac cgcctagagc ggagtgaacg 4921 gggaggggac cgtggggacc ggcttgatcg tgcgcggaca cctgctacca agcggagctt 4981 cagcaaggaa gtggaggagc ggagtagaga acggccctcc cagcctgagg ggctgcgcaa 5041 ggcagctagc ctcacggagg atcgggaccg tgggcgggat gccgtgaagc gagaagctgc 5101 cctaccccca gtgagccccc tgaaggcggc tctctctgag gaggagttag agaagaaatc 5161 caaggctatc attgaggaat atctccatct caatgacatg aaagaggcag tccagtgcgt 5221 gcaggagctg gcctcaccct ccttgctctt catctttgta cggcatggtg tcgagtctac 5281 gctggagcgc agtgccattg ctcgtgagca tatggggcag ctgctgcacc agctgctctg 5341 tgctgggcat ctgtctactg ctcagtacta ccaagggttg tatgaaatct tggaattggc 5401 tgaggacatg gaaattgaca tcccccacgt gtggctctac ctagcggaac tggtaacacc 5461 cattctgcag gaaggtgggg tgcccatggg ggagctgttc agggagatta caaagcctct 5521 gagaccgttg ggcaaagctg cttccctgtt gctggagatc ctgggcctcc tgtgcaaaag 5581 catgggtcct aaaaaggtgg ggacgctgtg gcgagaagcc gggcttagct ggaaggaatt 5641 tctacctgaa ggccaggaca ttggtgcatt cgtcgctgaa cagaaggtgg agtataccct 5701 gggagaggag tcggaagccc ctggccagag ggcactcccc tccgaggagc tgaacaggca 5761 gctggagaag ctgctgaagg agggcagcag taaccagcgg gtgttcgact ggatagaggc 5821 caacctgagt gagcagcaga tagtatccaa cacgttagtt cgagccctca tgacggctgt 5881 ctgctattct gcaattattt ttgagactcc cctccgagtg gacgttgcag tgctgaaagc 5941 gcgagcgaag ctgctgcaga aatacctgtg tgacgagcag aaggagctac aggcgctcta 6001 cgccctccag gcccttgtag tgaccttaga acagcctccc aacctgctgc ggatgttctt 6061 tgacgcactg tatgacgagg acgtggtgaa ggaggatgcc ttctacagtt gggagagtag 6121 caaggacccc gctgagcagc agggcaaggg tgtggccctt aaatctgtca cagccttctt 6181 caagtggctc cgtgaagcag aggaggagtc tgaccacaac tgaaagcttc gaattctgca 6241 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat 6301 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg 6361 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac 6421 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 6481 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt 6541 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 6601 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt 6661 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 6721 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag 6781 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 6841 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 6901 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 6961 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 7021 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 7081 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt 7141 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 7201 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 7261 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 7321 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 7381 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 7441 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat 7501 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 7561 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 7621 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 7681 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 7741 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 7801 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 7861 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 7921 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 7981 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg 8041 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 8101 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 8161 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 8221 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 8281 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg 8341 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 8401 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 8461 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct 8521 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc 8581 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 8641 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 8701 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt 8761 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 8821 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 8881 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 8941 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 9001 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 9061 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 9121 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 9181 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 9241 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 9301 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 9361 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 9421 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 9481 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 9541 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg 9601 cat //

pLB(N)CX-FLAG-CH3 Del33

133724

Retroviral vector with CMV promoter driving mutant form of CH3 domain fragment of human p300 (EP300), with N-terminal FLAG tag.

James A. DeCaprio

Borger et al J Virol. 2006 May . 80(9):4292-303.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/276246/718dd2d6-4a98-4573-8337-4362b748cb21/addgene-plasmid-133724-sequence-276246.gbk

full

LOCUS sequence-276246- 6610 bp DNA circular SYN 26-AUG-2024 DEFINITION Retroviral vector with CMV promoter driving mutant form of CH3 domain fragment of human p300 (EP300), with N-terminal FLAG tag.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6610) AUTHORS Borger DR, DeCaprio JA TITLE Targeting of p300/CREB binding protein coactivators by simian virus 40 is mediated through p53. JOURNAL J Virol. 2006 May . 80(9):4292-303. PUBMED 16611888 REFERENCE 2 (bases 1 to 6610) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Virol. 2006 May . 80(9):4292-303." FEATURES Location/Qualifiers source 1..6610 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 414..589 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 652..851 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1052..1468 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1398..1420 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1436..1452 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1511..1909 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" enhancer 2391..2770 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2771..2974 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2925..2945 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2971..2995 /label=LNCX /note="Human CMV promoter, forward primer" CDS 3089..3112 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" LTR 3512..4105 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(4208..4230) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 4316..4456 /label=bom /note="basis of mobility region from pBR322" primer_bind 4371..4390 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(4471..4488) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4642..5230) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4722..4741) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5401..6261) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6024..6043 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6262..6366) /gene="bla" /label=AmpR promoter primer_bind 6434..6452 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat 61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc 121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca 181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg 241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa 301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac 361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa 421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac 481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg 541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt 601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc 661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg 721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt 781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg 841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt 901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa 961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct 1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt 1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa 1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc 1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc 1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt 1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc 1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta 1441 tccagccctc actccttctc taggcgccgg aattccgatc tgatcaagag acaggatggg 1501 gtcgtttcgc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac 1561 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc 1621 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc 1681 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt 1741 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct 1801 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc 1861 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cgggactctg 1921 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 1981 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 2041 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccgggctcga tcccctcgcg 2101 agttggttca gctgctgcct gaggctggac gacctcgcgg agttctaccg gcagtgcaaa 2161 tccgtcggca tccaggaaac cagcagcggc tatccgcgca tccatgcccc cgaactgcag 2221 gagtggggag gcacgatggc cgctttggtc gaggcggatc cggccattag ccatattatt 2281 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatccata 2341 tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt 2401 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 2461 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 2521 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 2581 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 2641 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 2701 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 2761 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 2821 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 2881 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 2941 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg 3001 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg 3061 cggccccaag cttggatcct gaaccatgga ctacaaggac gacgatgaca agtcggccgc 3121 tggaggaggc cttggcttag atgatgagag caacaaccag caggctgcag ccacccagag 3181 cccaggcgat tctcgccgcc tgagtatcca gcagcggact ggtgtggttg ggcagcaaca 3241 gggcctccct tcccccactc ctgccactcc aacgacacca actggccaac agccaaccac 3301 cccgcagacg ccccagccca cttctcagcc tcagcctacc cctcccaata gcatgccacc 3361 ctacttgccc aggactcaag ctgctggccc tgtgtcccag ggtaaggcag caggccaggt 3421 gacccctcca acccctcctc agacttagga attcaagctt gttaacatcg ataaaataaa 3481 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca 3541 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga 3601 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg 3661 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc 3721 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc 3781 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc 3841 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct 3901 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg 3961 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg 4021 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac 4081 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca 4141 gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg 4201 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg 4261 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag 4321 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga 4381 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 4441 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 4501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 4561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 4981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5041 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 5101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5701 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 5821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 5881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 5941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6001 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct 6061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6361 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 6421 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat tcataccaga 6481 tcaccgaaaa ctgtcctcca aatgtgtccc cctcacactc ccaaattcgc gggcttctgc 6541 ctcttagacc actctaccct attccccaca ctcaccggag ccaaagccgc ggcccttccg 6601 tttctttgct //

pFusX1-24

63242

Encodes RVD positions 1-3 in TALEN assembly

Stephen Ekker

Ma et al Hum Gene Ther. 2016 Jun;27(6):451-63. doi: 10.1089/hum.2015.172

https://media.addgene.org/snapgene-media/v2.0.0/sequences/114298/b213a366-da4e-4f15-a6fe-310232f4486a/addgene-plasmid-63242-sequence-114298.gbk

partial

LOCUS sequence-114298- 694 bp DNA linear UNA 26-AUG-2024 DEFINITION Encodes RVD positions 1-3 in TALEN assembly. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 694) AUTHORS Ma AC, McNulty MS, Tanya PL, Jarryd CM, Martinez-Galvez G, Argue DP, Lee HB, Urban MD, Bullard CE, Blackburn PR, Man TK, Clark KJ, Ekker SC TITLE FusX: A rapid one-step TALE assembly system for genome science. JOURNAL Hum Gene Ther. 2016 Jun;27(6):451-63. doi: 10.1089/hum.2015.172 PUBMED 26854857 REFERENCE 2 (bases 1 to 694) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1089/hum.2015"; journalName: "Hum Gene Ther"; date: "2016-06"; volume: "27"; issue: "6"; pages: "451-63" FEATURES Location/Qualifiers source 1..694 /mol_type="genomic DNA" /organism="unspecified" primer_bind complement(398..415) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(398..414) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(407..429) /label=M13/pUC Forward /note="In lacZ gene" terminator 478..564 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 656..683 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" ORIGIN 1 tcacccggca accttgggta gcagcgaagt cgaggcattt ctgtcctggc tggtctagac 61 gtctcgtcag gccatggtcc tggcacagca ccggcaacag ccgctgcacc gtttcgagcg 121 cttgcttgcc gccaccgttg ctggcgatag ccaccacttg gtccggggtc aggccatggt 181 cctggcacag caccggcaac agccgctgca ccgtttcgag cgcttgcttg ccgccatcgt 241 ggctggcgat agccaccact tggtccggag tcaggccatg gtcctggcac agcaccggca 301 acagccgctg caccgtttcg agcgcttgct tgccgccatc gtggctggcg atagccacca 361 cttggtccgg ggtcaggttc agggggagac gcttaagact ggccgtcgtt ttacaacgtc 421 gtgactggga aaacatccat gctagcgtta acgcgagagt agggaactgc caggcatcaa 481 ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg 541 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg 601 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag 661 gccatcctga cggatggcct ttttgcgttt ctac //

pTUU

159749

Ubiquitin promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target sequence (U6-26 promoter). Selection of transgenic seeds by mTomato fluoresce.

Wolfgang Lukowitz

Angulo et al BioRxiv

https://media.addgene.org/snapgene-media/v2.0.0/sequences/313818/4f970a3e-6b98-49a6-bbed-418085c73209/addgene-plasmid-159749-sequence-313818.gbk

full

LOCUS sequence-313818- 17426 bp DNA circular SYN 25-AUG-2024 DEFINITION Ubiquitin promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target sequence (U6-26 promoter). Selection of transgenic seeds by mTomato fluoresce.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 17426) AUTHORS Angulo J, Astin CP, Bauer O , Blash KJ, Bowen NM, Chukwudinma NJ, Dinofrio AS, Faletti DO, Ghulam AM, Gusinde-Duffy CM, Horace KJ, Ingram AM, Isaack KE, Kiser RJ, Kobylanski JS, Long MR, Manning GA, Morales JM, Nguyen KH, Pham RT, Phillips MH, Reel TW, Seo JE, Vo HD, Wukuson AM, Yeary KA, Zheng GY, Lukowitz W TITLE Targeted mutagenesis of the Arabidopsis GROWTH-REGULATING FACTOR (GRF) gene family suggests competition of multiplexed sgRNAs for Cas9 apoprotein JOURNAL BioRxiv REFERENCE 2 (bases 1 to 17426) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BioRxiv" FEATURES Location/Qualifiers source 1..17426 /mol_type="other DNA" /organism="synthetic DNA construct" misc_RNA 1..76 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS 2277..2342 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 2349..2369 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2394..6494 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 2394..6494 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" RBS 6407..6415 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS 6495..6542 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 8033..8731 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein (Shaner et al., 2004)" /label=dTomato /note="mammalian codon-optimized" /translation="MSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH LFLYGMDELYK" primer_bind 8431..8450 /label=mCherry-F /note="mCherry, forward primer" primer_bind 8625..8648 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 8996..9170 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" misc_feature 9248..9272 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 9697..10491 /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" rep_origin 10578..11166 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11067..11086 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11320..11337 /label=L4440 /note="L4440 vector, forward primer" misc_feature 11352..11492 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(11418..11437) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 11578..11600 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 11836..12030 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(12096..13169) /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" CDS complement(13598..14227) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" misc_feature 15528..15552 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" primer_bind 15740..15762 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 15754..15771 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 15755..15771 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 16484..17275 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" ORIGIN 1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt 61 ggcaccgagt cggtgctttt ttttgcaaaa ttttccagat cgatttcttc ttcctctgtt 121 cttcggcgtt caatttctgg ggttttctct tcgttttctg taactgaaac ctaaaatttg 181 acctaaaaaa aatctcaaat aatatgattc agtggttttg tacttttcag ttagttgagt 241 tttgcagttc cgatgagata aaccaatacc atggatacac agtctagctc aacagagctt 301 ttaacccaaa ttggtacaat agaatacaac tttagatcat aattctcaaa agaaagagat 361 tccttagcta ttctatctgc cactccattt ccttctcggc ttgtatgcac aagcataaaa 421 tcctcaaact tgctaagtag atactttatg tcttggataa ttggattgag acttgacaag 481 cataactttc atgtaaccaa agacacaagt tgctgagaat ccacctcaaa aatgatcttc 541 ctataattga atcgggataa tgacagcaca gcccatctaa gagcctccac ttctacttcc 601 agcacgcttc ttacttttac cacagctctt gcacctaacc ataacacctt ccctgtatga 661 tcgcgaagca cccaccctaa gccacatttt aatccttctg ttggccatgc cccatcaaag 721 ttgcacttaa cccaagattg tggtggagct tcccatgttt ctcgtctgtc ccgacggtgt 781 tgtggttggt gctttcctta cattctgagc ctctttcctt ctaatccact catctgcatc 841 ttcttgtgtc cttactaata cctcattggt tccaaattcc ctccctttaa gcaccagctc 901 gtttctgttc ttccacagcc tcccaagtat ccaagggact aaagcctcca cattcttcag 961 atcaggatat tcttgtttaa gatgttgaac tctatggagg tttgtatgaa ctgatgatct 1021 aggaccggat aagttccctt cttcatagcg aacttattca aagaatgttt tgtgtatcat 1081 tcttgttaca ttgttattaa tgaaaaaata ttattggtca ttggactgaa cacgagtgtt 1141 aaatatggac caggccccaa ataagatcca ttgatatatg aattaaataa caagaataaa 1201 tcgagtcacc aaaccacttg ccttttttaa cgagacttgt tcaccaactt gatacaaaag 1261 tcattatcct atgcaaatca ataatcatac aaaaatatcc aataacacta aaaaattaaa 1321 agaaatggat aatttcacaa tatgttatac gataaagaag ttacttttcc aagaaattca 1381 ctgattttat aagcccactt gcattagata aatggcaaaa aaaaacaaaa aggaaaagaa 1441 ataaagcacg aagaattcta gaaaatacga aatacgcttc aatgcagtgg gacccacggt 1501 tcaattattg ccaattttca gctccaccgt atatttaaaa aataaaacga taatgctaaa 1561 aaaatataaa tcgtaacgat cgttaaatct caacggctgg atcttatgac gaccgttaga 1621 aattgtggtt gtcgacgagt cagtaataaa cggcgtcaaa gtggttgcag ccggcacaca 1681 cgagtcgtgt ttatcaactc aaagcacaaa tacttttcct caacctaaaa ataaggcaat 1741 tagccaaaaa caactttgcg tgtaaacaac gctcaataca cgtgtcattt tattattagc 1801 tattgcttca ccgccttagc tttctcgtga cctagtcgtc ctcgtctttt cttcttcttc 1861 ttctataaaa caatacccaa agagctcttc ttcttcacaa ttcagatttc aatttctcaa 1921 aatcttaaaa actttctctc aattctctct accgtgatca aggtaaattt ctgtgttcct 1981 tattctctca aaatcttcga ttttgttttc gttcgatccc aatttcgtat atgttctttg 2041 gtttagattc tgttaatctt agatcgaaga cgattttctg ggtttgatcg ttagatatca 2101 tcttaattct cgattagggt ttcatagata tcatccgatt tgttcaaata atttgagttt 2161 tgtcgaataa ttactcttcg atttgtgatt tctatctaga tctggtgtta gtttctagtt 2221 tgtgcgatcg aatttgtcga ttaatctgag tttttctgat taacaggttc tagatggatt 2281 acaaggacca cgacggggat tacaaggacc acgacattga ttacaaggat gatgatgaca 2341 agatggctcc gaagaagaag aggaaggttg gcatccacgg ggtgccagct gctgacaaga 2401 agtactcgat cggcctcgat attgggacta actctgttgg ctgggccgtg atcaccgacg 2461 agtacaaggt gccctcaaag aagttcaagg tcctgggcaa caccgatcgg cattccatca 2521 agaagaatct cattggcgct ctcctgttcg acagcggcga gacggctgag gctacgcggc 2581 tcaagcgcac cgcccgcagg cggtacacgc gcaggaagaa tcgcatctgc tacctgcagg 2641 agattttctc caacgagatg gcgaaggttg acgattcttt cttccacagg ctggaggagt 2701 cattcctcgt ggaggaggat aagaagcacg agcggcatcc aatcttcggc aacattgtcg 2761 acgaggttgc ctaccacgag aagtacccta cgatctacca tctgcggaag aagctcgtgg 2821 actccacaga taaggcggac ctccgcctga tctacctcgc tctggcccac atgattaagt 2881 tcaggggcca tttcctgatc gagggggatc tcaacccgga caatagcgat gttgacaagc 2941 tgttcatcca gctcgtgcag acgtacaacc agctcttcga ggagaacccc attaatgcgt 3001 caggcgtcga cgcgaaggct atcctgtccg ctaggctctc gaagtctcgg cgcctcgaga 3061 acctgatcgc ccagctgccg ggcgagaaga agaacggcct gttcgggaat ctcattgcgc 3121 tcagcctggg gctcacgccc aacttcaagt cgaatttcga tctcgctgag gacgccaagc 3181 tgcagctctc caaggacaca tacgacgatg acctggataa cctcctggcc cagatcggcg 3241 atcagtacgc ggacctgttc ctcgctgcca agaatctgtc ggacgccatc ctcctgtctg 3301 atattctcag ggtgaacacc gagattacga aggctccgct ctcagcctcc atgatcaagc 3361 gctacgacga gcaccatcag gatctgaccc tcctgaaggc gctggtcagg cagcagctcc 3421 ccgagaagta caaggagatc ttcttcgatc agtcgaagaa cggctacgct gggtacattg 3481 acggcggggc ctctcaggag gagttctaca agttcatcaa gccgattctg gagaagatgg 3541 acggcacgga ggagctgctg gtgaagctca atcgcgagga cctcctgagg aagcagcgga 3601 cattcgataa cggcagcatc ccacaccaga ttcatctcgg ggagctgcac gctatcctga 3661 ggaggcagga ggacttctac cctttcctca aggataaccg cgagaagatc gagaagattc 3721 tgactttcag gatcccgtac tacgtcggcc cactcgctag gggcaactcc cgcttcgctt 3781 ggatgacccg caagtcagag gagacgatca cgccgtggaa cttcgaggag gtggtcgaca 3841 agggcgctag cgctcagtcg ttcatcgaga ggatgacgaa tttcgacaag aacctgccaa 3901 atgagaaggt gctccctaag cactcgctcc tgtacgagta cttcacagtc tacaacgagc 3961 tgactaaggt gaagtatgtg accgagggca tgaggaagcc ggctttcctg tctggggagc 4021 agaagaaggc catcgtggac ctcctgttca agaccaaccg gaaggtcacg gttaagcagc 4081 tcaaggagga ctacttcaag aagattgagt gcttcgattc ggtcgagatc tctggcgttg 4141 aggaccgctt caacgcctcc ctggggacct accacgatct cctgaagatc attaaggata 4201 aggacttcct ggacaacgag gagaatgagg atatcctcga ggacattgtg ctgacactca 4261 ctctgttcga ggaccgggag atgatcgagg agcgcctgaa gacttacgcc catctcttcg 4321 atgacaaggt catgaagcag ctcaagagga ggaggtacac cggctggggg aggctgagca 4381 ggaagctcat caacggcatt cgggacaagc agtccgggaa gacgatcctc gacttcctga 4441 agagcgatgg cttcgcgaac cgcaatttca tgcagctgat tcacgatgac agcctcacat 4501 tcaaggagga tatccagaag gctcaggtga gcggccaggg ggactcgctg cacgagcata 4561 tcgcgaacct cgctggctcg ccagctatca agaaggggat tctgcagacc gtgaaggttg 4621 tggacgagct ggtgaaggtc atgggcaggc acaagcctga gaacatcgtc attgagatgg 4681 cccgggagaa tcagaccacg cagaagggcc agaagaactc acgcgagagg atgaagagga 4741 tcgaggaggg cattaaggag ctggggtccc agatcctcaa ggagcacccg gtggagaaca 4801 cgcagctgca gaatgagaag ctctacctgt actacctcca gaatggccgc gatatgtatg 4861 tggaccagga gctggatatt aacaggctca gcgattacga cgtcgatcat atcgttccac 4921 agtcattcct gaaggatgac tccattgaca acaaggtcct caccaggtcg gacaagaacc 4981 ggggcaagtc tgataatgtt ccttcagagg aggtcgttaa gaagatgaag aactactggc 5041 gccagctcct gaatgccaag ctgatcacgc agcggaagtt cgataacctc acaaaggctg 5101 agaggggcgg gctctctgag ctggacaagg cgggcttcat caagaggcag ctggtcgaga 5161 cacggcagat cactaagcac gttgcgcaga ttctcgactc acggatgaac actaagtacg 5221 atgagaatga caagctgatc cgcgaggtga aggtcatcac cctgaagtca aagctcgtct 5281 ccgacttcag gaaggatttc cagttctaca aggttcggga gatcaacaat taccaccatg 5341 cccatgacgc gtacctgaac gcggtggtcg gcacagctct gatcaagaag tacccaaagc 5401 tcgagagcga gttcgtgtac ggggactaca aggtttacga tgtgaggaag atgatcgcca 5461 agtcggagca ggagattggc aaggctaccg ccaagtactt cttctactct aacattatga 5521 atttcttcaa gacagagatc actctggcca atggcgagat ccggaagcgc cccctcatcg 5581 agacgaacgg cgagacgggg gagatcgtgt gggacaaggg cagggatttc gcgaccgtca 5641 ggaaggttct ctccatgcca caagtgaata tcgtcaagaa gacagaggtc cagactggcg 5701 ggttctctaa ggagtcaatt ctgcctaagc ggaacagcga caagctcatc gcccgcaaga 5761 aggactggga tccgaagaag tacggcgggt tcgacagccc cactgtggcc tactcggtcc 5821 tggttgtggc gaaggttgag aagggcaagt ccaagaagct caagagcgtg aaggagctgc 5881 tggggatcac gattatggag cgctccagct tcgagaagaa cccgatcgat ttcctggagg 5941 cgaagggcta caaggaggtg aagaaggacc tgatcattaa gctccccaag tactcactct 6001 tcgagctgga gaacggcagg aagcggatgc tggcttccgc tggcgagctg cagaagggga 6061 acgagctggc tctgccgtcc aagtatgtga acttcctcta cctggcctcc cactacgaga 6121 agctcaaggg cagccccgag gacaacgagc agaagcagct gttcgtcgag cagcacaagc 6181 attacctcga cgagatcatt gagcagattt ccgagttctc caagcgcgtg atcctggccg 6241 acgcgaatct ggataaggtc ctctccgcgt acaacaagca ccgcgacaag ccaatcaggg 6301 agcaggctga gaatatcatt catctcttca ccctgacgaa cctcggcgcc cctgctgctt 6361 tcaagtactt cgacacaact atcgatcgca agaggtacac aagcactaag gaggtcctgg 6421 acgcgaccct catccaccag tcgattaccg gcctctacga gacgcgcatc gacctgtctc 6481 agctcggggg cgacaagcgg ccagcggcga cgaagaaggc ggggcaggcg aagaagaaga 6541 agtgagctca gagctttcgt tcgtatcatc ggtttcgaca acgttcgtca agttcaatgc 6601 atcagtttca ttgcgcacac accagaatcc tactgagttt gagtattatg gcattgggaa 6661 aactgttttt cttgtaccat ttgttgtgct tgtaatttac tgtgtttttt attcggtttt 6721 cgctatcgaa ctgtgaaatg gaaatggatg gagaagagtt aatgaatgat atggtccttt 6781 tgttcattct caaattaata ttatttgttt tttctcttat ttgttgtgtg ttgaatttga 6841 aattataaga gatatgcaaa cattttgttt tgagtaaaaa tgtgtcaaat cgtggcctct 6901 aatgaccgaa gttaatatga ggagtaaaac acttgtagtt gtaccattat gcttattcac 6961 taggcaacaa atatattttc agacctagaa aagctgcaaa tgttactgaa tacaagtatg 7021 tcctcttgtg ttttagacat ttatgaactt tcctttatgt aattttccag aatccttgtc 7081 agattctaat cattgcttta taattatagt tatactcatg gatttgtagt tgagtatgaa 7141 aatatttttt aatgcatttt atgacttgcc aattgattga caacgaattc gggcccattt 7201 aaataggcga aaggggttta gggtttaagc tcagacttct attggagtaa atgggacggt 7261 gtcacatttt ccgttttgga aatgaacttt gggctcacgt tatgggctat tagatatttg 7321 atgggctttc tagtaaatac aatataagtt attgggctta gtttaaataa gcccatgttg 7381 gaaatatttg acacatgtct tggctactag tgctaaacat gcaaccgaac agttgtcgag 7441 acaagtcgca gcatatacaa tggatcaaac acgcctagtg tcgccgcgtc tcgctcatgt 7501 gtcaccttgt ttcctcgttt ttttttaatt tttcataagt tcttttgttt tatcttcaat 7561 acaaattttt ggctgtatct tgcaaactct tcgatcatat cgccaatata cgtgaacact 7621 ggtgatctaa tttgttgtgt taattgttaa atttagattc tattctccgg tttaaaagtg 7681 aattatatgt atcatggtta aaacattgta agtaagatga taataaaatg ataaatttag 7741 ttgatggata acgtgaagca aaaaatgaga tagatacatt tgattttgtc gtattttgac 7801 atatgcggag agtgagctac gcgcatgaag atcaagagac acttgctcga gctcacagag 7861 tgacgtgtaa aaagcttaga ctgaagtccc catgcaaacc taatcctacg tggctcaaac 7921 cacgagctca cttgacaata tataaactcc tcctaagtcc cgttctcttc atccatctct 7981 cacaacaaac aaaaagaaaa tggttaagct cagcaatctc ctcgttggat ctatgagcaa 8041 gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa 8101 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac 8161 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc 8221 ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa 8281 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg 8341 tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa 8401 gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg 8461 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca 8521 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc 8581 caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc 8641 ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct 8701 gttcctgtac ggcatggacg agctgtacaa gcttaagaag aagaaattca agcgtgaaga 8761 gcaagcaagg aaagctaagg tgaacaatga gaaaaagacg gaaatagtga aaccagagag 8821 ttgtagcaat gaaggagatg tcaaggatct gaaaagaaag gactctgagg atggaaacga 8881 gggtgaggaa gaagaagctt cttcgaaacc gaaaaagcca aaagttgctc tttctcatct 8941 tcaggacatt gacgacacag aagctgatca agaagaagag taatctagtc tcgagtttct 9001 ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct 9061 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc 9121 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc ccccgaatta 9181 attcggcgtt aattcagtac attaaaaacg tccgcaatgt gttattaagt tgtctaagcg 9241 tcaatttgtt tacaccacaa tatatcctgc caccagccag ccaacagctc cccgaccggc 9301 agctcggcac aaaatcacca ctcgatacag gcagcccatc agtccgggac ggcgtcagcg 9361 ggagagccgt tgtaaggcgg cagactttgc tcatgttacc gatgctattc ggaagaacgg 9421 caactaagct gccgggtttg aaacacggat gatctcgcgg agggtagcat gttgattgta 9481 acgatgacag agcgttgctg cctgtgatca ccgcggtttc aaaatcggct ccgtcgatac 9541 tatgttatac gccaactttg aaaacaactt tgaaaaagct gttttctggt atttaaggtt 9601 ttagaatgca aggaacagtg aattggagtt cgtcttgtta taattagctt cttggggtat 9661 ctttaaatac tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag aatatcaccg 9721 gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct 9781 gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc 9841 cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa 9901 ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg 9961 ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct 10021 gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg 10081 gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat 10141 aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag acactccatt taaagatccg 10201 cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac 10261 ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat 10321 cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgatc 10381 agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag 10441 cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta gtacctagaa 10501 tgcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 10561 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 10621 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 10681 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 10741 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 10801 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 10861 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 10921 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 10981 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 11041 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 11101 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 11161 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 11221 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 11281 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 11341 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 11401 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc 11461 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg 11521 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 11581 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagggtgcct 11641 tgatgtgggc gccggcggtc gagtggcgac ggcgcggctt gtccgcgccc tggtagattg 11701 cctggccgta ggccagccat ttttgagcgg ccagcggccg cgataggccg acgcgaagcg 11761 gcggggcgta gggagcgcag cgaccgaagg gtaggcgctt tttgcagctc ttcggctgtg 11821 cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat aagttttaaa 11881 gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta catgtgtgac 11941 cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc aatgtacggc 12001 tttgggttcc caatgtacgt gctatccaca ggaaacagac cttttcgacc tttttcccct 12061 gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga tgcttcgccc 12121 tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc ctcctccttc 12181 aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa acagaacttc 12241 ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca tctggcttct 12301 gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc 12361 aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt gatctcgcgg 12421 tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc gctctttacg 12481 atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg gccgttcttg 12541 gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca ggtttctacc 12601 aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac gtccgcaacg 12661 tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg gttcatggat 12721 tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact ggccatgccg 12781 gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat cacctcgcca 12841 gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct gcgactatcg 12901 cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc gcccttgggc 12961 ggcttcctaa tcgacggcgc accggctgcc ggcggttgcc gggattcttt gcggattcga 13021 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg 13081 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt 13141 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc 13201 cattgcaggg ccggcaggca acccagccgc ttacgcctgg ccaaccgccc gttcctccac 13261 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt 13321 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg 13381 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc 13441 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc 13501 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg 13561 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt 13621 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg 13681 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa 13741 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga 13801 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct 13861 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa 13921 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc 13981 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc 14041 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac 14101 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg 14161 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa 14221 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac 14281 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt 14341 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag 14401 gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc 14461 gatcatctcc gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg 14521 cggtttcatg cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg 14581 gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg 14641 ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct 14701 ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg 14761 agggtagggc gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg 14821 gtgcggtcga tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc 14881 atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg 14941 ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg 15001 tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc 15061 tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc 15121 gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag 15181 cccagcaggc cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa 15241 gtattctact ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg 15301 gcagcctgtc gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact 15361 gaacgtcaga agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc 15421 cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac 15481 gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa 15541 tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatccaag 15601 ctcaagctgc tctagcattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 15661 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt 15721 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt 15781 cgacttgcct tccgcacaat acatcatttc ttcttagctt tttttcttct tcttcgttca 15841 tacagttttt ttttgtttat cagcttacat tttcttgaac cgtagctttc gttttcttct 15901 ttttaacttt ccattcggag tttttgtatc ttgtttcata gtttgtccca ggattagaat 15961 gattaggcat cgaaccttca agaatttgat tgaataaaac atcttcattc ttaagatatg 16021 aagataatct tcaaaaggcc cctgggaatc tgaaagaaga gaagcaggcc catttatatg 16081 ggaaagaaca atagtatttc ttatataggc ccatttaagt tgaaaacaat cttcaaaagt 16141 cccacatcgc ttagataaga aaacgaagct gagtttatat acagctagag tcgaagtagt 16201 gattgggaga ccaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta 16261 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc 16321 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc 16381 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga 16441 tgttacgcag cagggcagtc gccctaaaac aaagttaaac atcatggggg aagcggtgat 16501 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc 16561 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag 16621 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt 16681 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt 16741 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga 16801 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac 16861 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt 16921 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct 16981 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt 17041 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga 17101 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga 17161 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt 17221 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataatgtct 17281 agctagaaat tcgttcaagc cgacgccgct tcgcggcgcg gcttaactca agcgttagat 17341 gcactaagca cataattgct cacagccaaa ctatcaggtc aagtctgctt ttattatttt 17401 taagcgtgca taataagccg gtctcg //

pGEX4T3-GST-PolB(K206A/K244A/T304I)

177134

Bacterial vector for expression of an N-terminal GST fusion of PolB(K206A/K244A/T304I) with a TEV protease site located between the GST tag and PolB(K206A/K244A/T304I)

Robert Sobol

Fang et al Nucleic Acids Res. 2019 Jul 9;47(12):6269-6286. doi: 10.1093/nar/gkz293.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/350560/e3ace548-9ab7-4ef0-b4eb-ea3b41682cf6/addgene-plasmid-177134-sequence-350560.gbk

full

LOCUS sequence-350560- 5993 bp DNA circular SYN 26-AUG-2024 DEFINITION Bacterial vector for expression of an N-terminal GST fusion of PolB(K206A/K244A/T304I) with a TEV protease site located between the GST tag and PolB(K206A/K244A/T304I). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5993) AUTHORS Fang Q, Andrews J, Sharma N, Wilk A, Clark J, Slyskova J, Koczor CA, Lans H, Prakash A, Sobol RW TITLE Stability and sub-cellular localization of DNA polymerase beta is regulated by interactions with NQO1 and XRCC1 in response to oxidative stress. JOURNAL Nucleic Acids Res. 2019 Jul 9;47(12):6269-6286. doi: 10.1093/nar/gkz293. PUBMED 31287140 REFERENCE 2 (bases 1 to 5993) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: "Nucleic Acids Res"; date: "2019-07-9- 9"; volume: "47"; issue: "12"; pages: "6269-6286" FEATURES Location/Qualifiers source 1..5993 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 175..205 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 213..229 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 237..253 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 237..253 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 249..776 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR QPSEAVVWLCRS" primer_bind complement(269..286) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(269..285) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(278..300) /label=M13/pUC Forward /note="In lacZ gene" promoter 878..906 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 914..930 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." CDS 953..1606 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 1564..1586 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 1613..1630 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1631..1651 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" primer_bind complement(2738..2760) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2905..2923) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2991..3095 /gene="bla" /label=AmpR promoter CDS 3096..3956 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3314..3333) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4127..4715 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4616..4635 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4869..4886 /label=L4440 /note="L4440 vector, forward primer" promoter 4959..5036 /gene="lacI" /label=lacI promoter CDS join(5037..5993,1..126) /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(5056..5075) /label=LacI-R /note="LacI, reverse primer" ORIGIN 1 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc 61 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg 121 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca 181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg 241 aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg 301 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc 361 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg 421 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg 481 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg 541 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg 601 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag 661 gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca cggtgcacca 721 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca 781 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac 841 atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt 901 ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagtat tcatgtcccc 961 tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata 1021 tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa 1081 caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt 1141 taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg 1201 tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag 1261 atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct 1321 tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt 1381 aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt 1441 atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat 1501 tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt 1561 gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggttcc 1621 gcgtggatcc gaaaacctgt attttcaggg ctcgaattcc cgggtcgaca tgagcaaacg 1681 gaaggcgccg caggagactc tcaacggggg aatcaccgac atgctcacag aactcgcaaa 1741 ctttgagaag aacgtgagcc aagctatcca caagtacaat gcttacagaa aagcagcatc 1801 tgttatagca aaatacccac acaaaataaa gagtggagct gaagctaaga aattgcctgg 1861 agtaggaaca aaaattgctg aaaagattga tgagttttta gcaactggaa aattacgtaa 1921 actggaaaag attcggcagg atgatacgag ttcatccatc aatttcctga ctcgagttag 1981 tggcattggt ccatctgctg caaggaagtt tgtagatgaa ggaattaaaa cactagaaga 2041 tctcagaaaa aatgaagata aattgaacca tcatcagcga attgggctga aatattttgg 2101 ggactttgaa aaaagaattc ctcgtgaaga gatgttacaa atgcaagata ttgtactaaa 2161 tgaagttaaa aaagtggatt ctgaatacat tgctacagtc tgtggcagtt tcagaagagg 2221 tgcagagtcc agtggtgaca tggatgttct cctgacccat cccagcttca cttcagaatc 2281 aaccgcacag ccaaaactgt tacatcaggt tgtggagcag ttacaaaagg ttcattttat 2341 cacagatacc ctgtcaaagg gtgagacaaa gttcatgggt gtttgccagc ttcccagtgc 2401 aaatgatgaa aaagaatatc cacacagaag aattgatatc aggttgatac ccaaagatca 2461 gtattactgt ggtgttctct atttcactgg gagtgatatt ttcaataaga atatgagggc 2521 tcatgcccta gaaaagggtt tcacaatcaa tgagtacacc atccgtccct tgggagtcat 2581 tggagttgca ggagaacccc tgccagtgga tagtgaaaaa gacatctttg attacatcca 2641 gtggaaatac cgggaaccca aggaccggag cgaatgagcg gccgcatcgt gactgactga 2701 cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 2761 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 2821 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag 2881 tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca 2941 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 3001 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 3061 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 3121 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 3181 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 3241 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 3301 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac 3361 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 3421 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 3481 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 3541 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 3601 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc 3661 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 3721 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 3781 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc 3841 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 3901 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 3961 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 4021 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 4081 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 4141 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 4201 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 4261 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 4321 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 4381 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 4441 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 4501 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 4561 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 4621 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 4681 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 4741 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 4801 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 4861 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc 4921 ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa tggcgcaaaa 4981 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga 5041 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc 5101 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga 5161 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt 5221 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg 5281 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa 5341 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc 5401 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta 5461 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct 5521 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa 5581 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc 5641 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg 5701 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga 5761 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc 5821 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt 5881 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg 5941 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttg //

pGBW-m4046326

145561

Yeast Expression plasmid for SARS-CoV-2 ORF7a protein

SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/287738/b34264a7-03cb-4f97-8658-da95ba44ea45/addgene-plasmid-145561-sequence-287738.gbk

full

LOCUS sequence-287738- 5172 bp DNA circular SYN 26-AUG-2024 DEFINITION Yeast Expression plasmid for SARS-CoV-2 ORF7a protein. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5172) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5172) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5172 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 25..466 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 25..142 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 437..460 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 841..858 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 859..882 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(927..945) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1296..2099 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin 2182..3164 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3242..3271 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3293..3880 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator complement(3892..3919) /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3943..4134) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4135..4995) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4758..4777 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4996..5098) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator complement(5099..5130) /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg 61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg 121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat 181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta 241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag 301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt 361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta 421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg 481 aaaattatcc tattcttagc attgataacc ctggctacgt gtgagcttta tcactaccag 541 gaatgcgtta gggggacaac tgtcttgttg aaggaacctt gttcatctgg aacttacgaa 601 ggcaattcgc catttcatcc attggctgat aacaagttcg ccttaacctg ttttagtact 661 caattcgctt tcgcttgtcc cgacggtgtg aaacacgttt atcaattgag agcgagatcc 721 gtaagcccaa agttatttat ccgtcaagaa gaagttcaag agttgtactc cccaattttt 781 ctaattgtcg ccgctatcgt tttcattacc ttgtgtttca cactgaagcg aaagactgaa 841 ctcgtgccga gaggttcttg gagccatccg cagtttgaaa aataaacagg ccccttttcc 901 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc 961 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt 1021 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt 1081 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg 1141 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat 1201 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc 1261 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac 1321 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc 1381 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg 1441 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt 1501 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct 1561 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg 1621 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta 1681 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga 1741 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg 1801 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg 1861 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca 1921 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg 1981 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg 2041 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa 2101 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat 2161 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc 2221 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa 2281 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa 2341 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac 2401 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct 2461 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc 2521 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag 2581 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact 2641 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc 2701 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt 2761 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac 2821 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt 2881 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg 2941 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca 3001 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata 3061 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc 3121 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact 3181 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac 3241 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc 3301 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 3361 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc 3421 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc 3481 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 3541 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 3601 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 3661 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 3721 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 3781 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg 3841 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag 3901 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc 3961 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc 4021 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt 4081 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca 4141 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 4201 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc 4261 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 4321 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat 4381 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 4441 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 4501 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 4561 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt 4621 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac 4681 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 4741 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 4801 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc 4861 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 4921 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 4981 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac 5041 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag 5101 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa 5161 tggtcagtta ga //

Gr4-T2A-QF2 HDR plasmid

140944

Plasmid for CRISPR-generated knock-in line that expresses a QF2 transcriptional activator by knocking-in a T2A-QF2 fragment into the coding sequence of the endogenous Aedes aegypti Gr4 gene

Leslie Vosshall

Jove et al Neuron. 2020 Dec 23;108(6):1163-1180.e12. doi: 10.1016/j.neuron.2020.09.019. Epub 2020 Oct 12.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/313602/2d9602aa-c6a0-4e4d-8ecb-c59cad69cdf3/addgene-plasmid-140944-sequence-313602.gbk

full

LOCUS sequence-313602- 7630 bp DNA circular SYN 26-AUG-2024 DEFINITION Plasmid for CRISPR-generated knock-in line that expresses a QF2 transcriptional activator by knocking-in a T2A-QF2 fragment into the coding sequence of the endogenous Aedes aegypti Gr4 gene. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7630) AUTHORS Jove V, Gong Z, Hol FJH, Zhao Z, Sorrells TR, Carroll TS, Prakash M, McBride CS, Vosshall LB TITLE Sensory Discrimination of Blood and Floral Nectar by Aedes aegypti Mosquitoes. JOURNAL Neuron. 2020 Dec 23;108(6):1163-1180.e12. doi: 10.1016/j.neuron.2020.09.019. Epub 2020 Oct 12. PUBMED 33049200 REFERENCE 2 (bases 1 to 7630) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.neuron.2020.09.019"; journalName: "Neuron"; date: "2020-12-23- 23"; volume: "108"; issue: "6"; pages: "1163-1180.e12" FEATURES Location/Qualifiers source 1..7630 /mol_type="other DNA" /organism="synthetic DNA construct" regulatory 1092..1101 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 1098..1775 /codon_start=1 /product="improved tetrameric variant of DsRed fluorescent protein" /label=DsRed2 /note="mammalian codon-optimized" /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH LFL" primer_bind complement(1278..1298) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 1690..1713 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 1896..2017 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1933..1952) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1987..2006 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3559..3575) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3559..3575) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3572..3594) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3583..3599 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3607..3637) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3652..3673 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3790..3807) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3961..4549) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4041..4060) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4720..5580) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5343..5362 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5581..5685) /gene="bla" /label=AmpR promoter primer_bind 5753..5771 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5809..5831) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5931..5950 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6144..6166 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6158..6175 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6159..6175 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 7082..7135 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" ORIGIN 1 ctccatgccc gttgggctac tagccgtgtc aataagagca ttacccgcct cctccgtcag 61 ttggagctcc ctcctaccgc cacggctacg gcctcgataa tgccgcacgt gatggagcag 121 cctctcagta ccagcattaa ccccgtcaac gaccgcttca acggtattcc caaccccact 181 ccgtataact ccgatgcagc tctcgatgct atcactcaga ccaacgatta tggaagcgta 241 aatacacatg gtatcctctc tacttacccg ccaccggcta cgcaccttaa tgaagcttcc 301 gtcgctctcg ctcccggtgg cgcccccccc cgaccgcctc ctccgtatgt tgacagcacg 361 accaatcacc cgccgtacca ctcgaatctg gttccaatgg cgaactttgg ttactcgacc 421 gttgattacg atgccatggt tgacgatttg gctagcattg aatacacgga cgctgtggat 481 gtcgacccac agtttatgac caatctggga ttcgttcctg gatgtaactt ctccgacatt 541 aatacatacg aacagtgatg acgtctatcg ataccgtcga ctaaagccaa atagaaaatt 601 attcagttcc tggcttaagt ttttaaaagt gatattattt atttggttgt aaccaaccaa 661 aagaatgtaa ataactaata cataattatg ttagttttaa gttagcaaca aattgatttt 721 agctatatta gctacttggt taataaatag aatatattta tttaaagata attgcgtttt 781 tattgtcagg gagtgagttt gcttaaaaac tcgtttagat ccaattcgag ctcgcccggg 841 gatctaattc aattagagac taattcaatt agagctaatt caattaggat ccaagcttat 901 cgatttcgaa ccctcgaccg ccggagtata aatagaggcg cttcgtctac ggagcgacaa 961 ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc 1021 gcagctgaac aagctaaaca atcggggtac cgctagagtc gacggtaccg cgggcccggg 1081 atccaccggt cgccaccatg gcctcctccg agaacgtcat caccgagttc atgcgcttca 1141 aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc gagggcgagg 1201 gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc ggccccctgc 1261 ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg tacgtgaagc 1321 accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc aagtgggagc 1381 gcgtgatgaa cttcgaggac ggcggcgtgg cgaccgtgac ccaggactcc tccctgcagg 1441 acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc gacggccccg 1501 tgatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg 1561 gcgtgctgaa gggcgagacc cacaaggccc tgaagctgaa ggacggcggc cactacctgg 1621 tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc tactactacg 1681 tggacgccaa gctggacatc acctcccaca acgaggacta caccatcgtg gagcagtacg 1741 agcgcaccga gggccgccac cacctgttcc tgtagcggcc gcgactctag atcataatca 1801 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga 1861 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg 1921 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 1981 ctagttgtgg tttgtccaaa ctcatcaatg tatcttacag ggaaaactgg atccatgacg 2041 gatcgtttca cgatgcagtt tctggactgc tgattacggc ccaactgttc tcaataatgc 2101 cagtttgtgg aattggacag aaggatacta ctaagcttca tttctcttgg aaaagtaagc 2161 gcatcttcta ctcatacgcg gcatgtatgg gaactgcttt cctggcggtg actagtacga 2221 ttcgattcgt ggatagaaat ttcaactttt ctaggctgac gggagtcttt ttctatttct 2281 acaacctcta cggaatgtac tgtttcgtga gagttgcaca gaaatggccg gttctgatgc 2341 agaagtggtt taatgttgaa caactgctgc cgcagagttc caacattata gaacgtggaa 2401 agcttgccaa taagatcaag ctgatatcga ttcttgtgat cacgctctcc ctgatggaac 2461 atatgctgtc gattgtggca gccgtttact ataccccgaa ttgcccaaac atcaaggatc 2521 cggtcaaaat gttcttcaag tccaactttt tgttcgtatt ctactacttt gagtattcgg 2581 agattcgagg attcgtggtc aaatttatca acgtgatcag cacctttgtt tggagttaca 2641 tagatttgtt cgtcattatc gtaagcattg gactatctca cacgttccgc aggatcaaca 2701 accatctgat gaatcataaa cgagaggtac gaagttcaga acattaaacc gtagattact 2761 aaacaatgtg cccttccaga aaatgaccga acagttttgg ggtgagcaac gtcaaaacta 2821 tcgaaatatt tgtgaccttg ttcggttcgt tgacgacgct atttcgatca ttaccatgct 2881 atcgatttca aacaacctgt tcttcatctg cgcctcaatt cttaacagtt tgaagtattg 2941 tccaatccga ttccttcata atacacactg gttgatatct ctgtcttcat tccagcaccc 3001 acccaacctt ggtccataca gtgtacttct ggttcgggct ggccttcctg attggacgaa 3061 ctttggccgt gtccatgtgc accgcagcag tcaacgatga atcgcagcgt ccgtttgaag 3121 ttcttcgggc tattccccgt gacggatggt gcgtggaagc gaaacgcttt gccgaagagg 3181 tcatcaacga tacggtggcc ctcaccggga tgaagttctt caacatgacg cgcaaactgg 3241 tgctcaaagt cacgggttcg atcatcacct acgaactggt gctgatccag tttcaccagg 3301 acgagacggc cgattacgat ctgtgcacgt ttaggcgtac gtaggtgaag cggattaatg 3361 ggagatgaat aaatatgttg cattcagcat taaaataata ttaatttaga ataggttttc 3421 aatcgaatga atttgttttt atgttcagca catcgtacac acttggataa ttctaatttc 3481 tgtaaccttt ataattctac tgaggctcca aatacaccta gagtcgacct gcaggcatgc 3541 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 3601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 3661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 3721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 3901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 3961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 4021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 4081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 4141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 4201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 4261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 4321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 4381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 4441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 5701 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 5761 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 5821 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 5881 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 5941 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 6001 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 6061 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 6121 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt 6181 cgagctcggt acccgggact ctcctaaaat ctcaagtata cataaagtca cgatgatttc 6241 actttcaaac tattcactgt ttccattaga gtctgggtct cgctgctttt taaatatcct 6301 tttaataaat aattgcactc acattcgtta actgaaatca atttattctt atttcaaaag 6361 ctgttccaac atggcccttc cacgcgtatt gacatggttt ccatagaaat tcccaaaaac 6421 acacacatca aacccaaatc tagacccact tgacgattcg ctattaatcc gactgaaacc 6481 atgtagtggg ctgcaacaaa gtggtaagcc aacccaccga tggagtcttt tatcaggaaa 6541 tcaacagttg attcatatct acctttccgc agtcgtctcc tctttattat attccgccaa 6601 cacgaaacgg ccacggagtt ctggttttac tcggtaatta ttactgcacg gtatgccaaa 6661 tggttggctg ctgcattaaa cacctttcgt gttaagagtc aggttgtttt tccatacaga 6721 cttcctagct atttaataca gctgtttgtt ttcccatgct tctctccggg actgcaccaa 6781 caatgaactg catatactgt aggattactc cctttagttt tatcctgtgg gttaaccgat 6841 tgtgaggatc aaaaggatga tgcgggttag ccaattgtcg accatggcga gaaaattgct 6901 tccccaggtg ttggaacccc gacaaacagg aaagatagtg aggtaaaatg tgtaacaatt 6961 aacatttaaa gtcaaaattt tatcaactcc catggattca aggtttaacg gcgagttggt 7021 ccccgtccaa agtaagagtc gagcttttct gaagcacttc aagtatccca aacgtgcagg 7081 agagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgcc 7141 acccaagcgc aaaacgctta acgctgcggc tgaggctaac gctcatgccg acggacacgc 7201 cgacggaaac gccgacggac acgtggccaa tacggccgcg tcctcgaata atgcgaggtt 7261 cgctgatctc actaacatcg atactccggg tctgggaccc acaactacga ccctgctcgt 7321 ggaaccagca cgctcaaagc gtcaacgagt gtcccgcgca tgcgaccagt gccgtgcagc 7381 ccgagagaaa tgcgacggaa tacagcctgc gtgtttcccg tgcgtttccc agggaaggtc 7441 ctgcacttat caggcttcgc cgaaaaagag gggagttcaa accggttata ttcgtacgct 7501 ggagctcgcc ctcgcctgga tgtttgaaaa tgtcgcgcgt tccgaagatg ccttgcataa 7561 cctcctcgtc cgtgacgccg gacaaggatc agctctgctc gttggtaaag attcgccggc 7621 tgccgagcga //

TAL3202

41376

Keith Joung

Sander et al Nat Biotechnol. 2011 Aug 5;29(8):697-8. doi: 10.1038/nbt.1934.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/56341/2329d975-bfe5-4e44-b18b-36eaf471ec1e/addgene-plasmid-41376-sequence-56341.gbk

partial

LOCUS sequence-56341-e 892 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 892) AUTHORS Sander JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung JK, Yeh JR TITLE Targeted gene disruption in somatic zebrafish cells using engineered TALENs. JOURNAL Nat Biotechnol. 2011 Aug 5;29(8):697-8. doi: 10.1038/nbt.1934. PUBMED 21822241 REFERENCE 2 (bases 1 to 892) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nbt"; journalName: "Nat Biotechnol"; date: "2011-08-5- 5"; volume: "29"; issue: "8"; pages: "697-8" FEATURES Location/Qualifiers source 1..892 /mol_type="genomic DNA" /organism="unspecified" ORIGIN 1 gcgctgctga ctgtggcggg tgagcttagg gggcctccgc tccagctcga caccgggcag 61 ctgctgaaga tcgcgaagag agggggagta acagcggtag aggcagtgca cgcctggcgc 121 aatgcgctca ccggggcccc cttgaacctg accccagacc aggtagtcgc aatcgcgaac 181 aataatgggg gaaagcaagc cctggaaacc gtgcaaaggt tgttgccggt cctttgtcaa 241 gaccacggcc ttacaccgga gcaagtcgtg gccattgcat cccacgacgg tggcaaacag 301 gctcttgaga cggttcagag acttctccca gttctctgtc aagcccacgg gctgactccc 361 gatcaagttg tagcgattgc gaataacaat ggagggaaac aagcattgga gactgtccaa 421 cggctccttc ccgtgttgtg tcaagcccac ggtttgacgc ctgcacaagt ggtcgccatc 481 gccaacaaca acggcggtaa gcaggcgctg gaaacagtac agcgcctgct gcctgtactg 541 tgccaggatc atggactgac cccagaccag gtagtcgcaa tcgcgaacaa taatggggga 601 aagcaagccc tggaaaccgt gcaaaggttg ttgccggtcc tttgtcaaga ccacggcctt 661 acaccggagc aagtcgtggc cattgcaagc aacatcggtg gcaaacaggc tcttgagacg 721 gttcagagac ttctcccagt tctctgtcaa gcccacgggc tgactcccga tcaagttgta 781 gcgattgcgt cgcatgacgg agggaaacaa gcattggaga ctgtccaacg gctccttccc 841 gtgttgtgtc aagcccacgg tttgacgcct gcacaagtgg tcgccatcgc ca //

Syntaxin-4 d29

44190

Thomas Weimbs

Reales et al PLoS One. 2011;6(6):e21181. doi: 10.1371/journal.pone.0021181. Epub 2011 Jun 15.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/66276/38bc1a87-578c-4166-984e-c7beb63f8f89/addgene-plasmid-44190-sequence-66276.gbk

partial

LOCUS sequence-66276-e 912 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 912) AUTHORS Reales E, Sharma N, Low SH, Folsch H, Weimbs T TITLE Basolateral sorting of syntaxin 4 is dependent on its N-terminal domain and the AP1B clathrin adaptor, and required for the epithelial cell polarity. JOURNAL PLoS One. 2011;6(6):e21181. doi: 10.1371/journal.pone.0021181. Epub 2011 Jun 15. PUBMED 21698262 REFERENCE 2 (bases 1 to 912) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One."; date: "2011"; volume: "6(6):e21181. doi"; pages: " 10.1371/journal.pone.0021181. Epub 2011 Jun 15" FEATURES Location/Qualifiers source 1..912 /mol_type="genomic DNA" /organism="unspecified" protein_bind 11..29 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 32..50 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 60..84 /label=LNCX /note="Human CMV promoter, forward primer" ORIGIN 1 agcagagctc tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga 61 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat 121 agaagacacc gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc 181 gagctcggat ccactagtcc agtgtggtgg aattcgatat ggggacggca cggctgggga 241 gcccggacga ggagttcttc cacaaggtcc ggacaattcg gcagactatt gtcaaactgg 301 ggaataaagt ccaggagttg gagaaacagc aggtcaccat cctggccacg ccccttcccg 361 aggagagcat gaagcaggag ctgcagaacc tgcgcgatga gatcaaacag ctggggaggg 421 agatccgcct gcagctgaag gccatagagc cccagaagga ggaagctgat gagaactata 481 actccgtcaa cacaagaatg agaaaaaccc agcatggggt cctgtcccag caattcgtgg 541 agctcatcaa caagtgcaat tcaatgcagt ccgaataccg ggagaagaac gtggagcgga 601 ttcggaggca gctgaagatc accaatgctg ggatggtgtc tgatgaggag ttggagcaga 661 tgctggacag tgggcaaagc gaggtgtttg tgtccaatat cctgaaggac acgcaggtga 721 ctcgacaggc cttaaatgag atctcggccc ggcacagtga gatccagcag cttgaacgca 781 gtattcgtga gctgcacgac atattcactt ttctggctac cgaagtggag atgcaggggg 841 agatgatcaa tcggattgag aagaacatcc tgagctcagc ggactacgtg gaacgtgggc 901 aggagcacgt ca //

pCAG-T3-hCasD10A-pA

51638

Low

Expresses Cas9-nickase in mammalian cells and zygotes.

Wataru Fujii

Fujii et al Biochem Biophys Res Commun. 2014 Jan 31. pii: S0006-291X(14)00176-4. doi: 10.1016/j.bbrc.2014.01.141.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/80999/bf3f7f77-4beb-4c94-96d0-c30b67b5c3ed/addgene-plasmid-51638-sequence-80999.gbk

partial

LOCUS sequence-80999-e 894 bp DNA linear UNA 26-AUG-2024 DEFINITION Expresses Cas9-nickase in mammalian cells and zygotes.. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 894) AUTHORS Fujii W, Onuma A, Sugiura K, Naito K TITLE Efficient generation of genome-modified mice via offset-nicking by CRISPR/Cas system. JOURNAL Biochem Biophys Res Commun. 2014 Jan 31. pii: S0006-291X(14)00176-4. doi: 10.1016/j.bbrc.2014.01.141. PUBMED 24491566 REFERENCE 2 (bases 1 to 894) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biochem Biophys Res Commun. 2014 Jan 31. pii: S0006-291X(14)00176-4. doi: 10.1016/j.bbrc.2014.01.141." FEATURES Location/Qualifiers source 1..894 /mol_type="genomic DNA" /organism="unspecified" primer_bind 34..53 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" promoter 83..101 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 162..185 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 195..215 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" ORIGIN 1 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc 61 atcattttgg caaagaattc gaaattaacc ctcactaaag ggaacaaaag ctggagctcc 121 accgcggtgg cggccgctct agcccgggcg gatccaccat ggattacaag gatgacgacg 181 ataagatcat ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg 241 acaagaagta ctccattggg ctcgccatcg gcacaaacag cgtcggctgg gccgtcatta 301 cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc gatcgccaca 361 gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg gccgaagcca 421 cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg atctgctacc 481 tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc cataggctgg 541 aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc tttggcaata 601 tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg aggaagaagc 661 ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg gcgcatatga 721 tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac agcgatgtcg 781 acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag aacccgatca 841 acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa tccc //

pTSA-13

17327

High

Takeshi Sano

Sano et al J Biol Chem. 1995 Nov 24. 270(47):28204-9.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/242416/5ff097b3-599c-4125-a60b-f6429651f59d/addgene-plasmid-17327-sequence-242416.gbk

full

LOCUS sequence-242416- 5036 bp DNA circular SYN 25-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5036) AUTHORS Sano T, Pandori MW, Chen X, Smith CL, Cantor CR TITLE Recombinant core streptavidins. A minimum-sized core streptavidin has enhanced structural stability and higher accessibility to biotinylated macromolecules. JOURNAL J Biol Chem. 1995 Nov 24. 270(47):28204-9. PUBMED 7499314 REFERENCE 2 (bases 1 to 5036) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Chem. 1995 Nov 24. 270(47):28204-9." FEATURES Location/Qualifiers source 1..5036 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 10..38 /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" terminator complement(404..451) /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 447..465 /label=T7 Term /note="T7 terminator, reverse primer" CDS complement(519..551) /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" RBS 958..980 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" primer_bind complement(1011..1030) /label=T7 /note="T7 promoter, forward primer" promoter complement(1012..1030) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1114..1133) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS 2591..2782 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind complement(2776..2798) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 2884..3023 /label=bom /note="basis of mobility region from pBR322" primer_bind 2939..2958 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3038..3055) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3209..3797) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3289..3308) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3968..4828) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4591..4610 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4829..4933) /gene="bla" /label=AmpR promoter primer_bind 5001..5019 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa 61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg 121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt 181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata 241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg 301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac 361 cacacccgtc ctgtggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga 421 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact 481 cagcttcctt tcgggctttg ttagcagccg gatccgcgac ccatttgctg tccaccagtc 541 atgctagcca tatgggcctt cttcgccgcg tcgatggagg cggcggacgg ctacaccttg 601 gtgaaggtgt cgtggccgac cagcgtggac ttccaggcgt tggcctcggt ggtgccggag 661 gtcagcagcc actgggtgtt gatcctcgcc tcggcgccgc cgacgtactg gccgctccac 721 gtggtcgcgg agtgggcgtt gcggtagtta ttcttccagg ccaccgtcca accgagggcg 781 gtgccgctgc cgtcggtggc cggggcgctg tcgtaacgac cggtcaggac gtagcggctc 841 tcggcgttgc cgacggccga ctcgtaggtt ccggtcaggg cgccgtcggc gcccgcggtc 901 acgatgaagg tcgagccgag ctggttgtac caggtgccgg tgatgcccat atgtatatct 961 ccttcttaaa gttaaacaaa attatttcta gagggaaacc gttgtggtct ccctatagtg 1021 agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg 1081 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg 1141 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag 1201 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg 1261 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg 1321 gagagcgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc 1381 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac 1441 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga 1501 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca 1561 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg 1621 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta 1681 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc 1741 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa 1801 cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga 1861 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc 1921 gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg 1981 attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac 2041 caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat 2101 ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac 2161 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg 2221 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt 2281 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt 2341 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac 2401 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag 2461 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt 2521 gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac 2581 ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag 2641 ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat 2701 ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg 2761 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 2821 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 2881 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg 2941 gcatcagagc agattgtact gagagtgcac cattgcggtg tgaaataccg cacagatgcg 3001 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct 3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 3601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 3721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 3781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 3841 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 3901 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 3961 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 4021 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 4081 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 4141 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 4201 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 4261 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg 4321 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 4381 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 4441 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 4501 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 4561 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa 4621 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 4681 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 4741 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 4801 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 4861 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 4921 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta 4981 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaa //

pBbA1c-RFP

35335

Jay Keasling

Lee et al J Biol Eng. 2011 Sep 20;5:12.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/43002/2d8c7486-0ed6-44dc-969b-8290908e6aaf/addgene-plasmid-35335-sequence-43002.gbk

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LOCUS sequence-43002-e 4038 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4038) AUTHORS Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD TITLE BglBrick vectors and datasheets: A synthetic biology platform for gene expression. JOURNAL J Biol Eng. 2011 Sep 20;5:12. PUBMED 21933410 REFERENCE 2 (bases 1 to 4038) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Eng."; date: "2011-09-20"; volume: "5"; pages: "12" FEATURES Location/Qualifiers source 1..4038 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 7..84 /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 85..1167 /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(104..123) /label=LacI-R /note="LacI, reverse primer" promoter 1399..1428 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 1436..1452 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." CDS 1493..2170 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 2203..2274 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2290..2317 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(2345..2362) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2479..3024) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(2559..2578) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator complement(3138..3232) /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3253..3912) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 3827..3846 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(3913..4015) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 gacgtcgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa 61 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat 121 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg 181 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg 241 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc 301 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc 361 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac 421 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat 481 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac 541 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag 601 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc 661 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg 721 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg 781 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc 841 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga 901 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat 961 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg 1021 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc 1081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 1141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt 1201 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca 1261 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct 1321 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa 1381 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg 1441 agcggataac aatttcagaa ttcaaaagat cttttaagaa ggagatatac atatggcgag 1501 tagcgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag gttccgttaa 1561 cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac 1621 cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc 1681 gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc cggactacct 1741 gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg 1801 tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct acaaagttaa 1861 actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg 1921 ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat 1981 gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca cctacatggc 2041 taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg acatcacctc 2101 ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac 2161 cggtgcttaa ggatccaaac tcgagtaagg atctccaggc atcaaataaa acgaaaggct 2221 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag 2281 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacctaggg atatattccg 2341 cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt 2401 acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag 2461 ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg 2521 acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc 2581 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct 2641 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct 2701 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta 2761 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg 2821 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg 2881 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct 2941 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt 3001 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag 3061 atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag 3121 ttgttactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg 3181 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag 3241 ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag 3301 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat 3361 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt 3421 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac 3481 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc 3541 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag 3601 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca 3661 tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg 3721 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa 3781 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa 3841 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat 3901 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc 3961 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc 4021 tcattttcgc cagatatc //

Lentiviral-His-tagged-PSPH

134786

Stable overexpression lentiviral vector of His tagged PSPH

Seon-Young Kim

Park et al Mol Cells. 2019 Aug 31;42(8):604-616. doi: 10.14348/molcells.2019.0160.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/266694/eddea159-4560-452b-bac0-291c3920a375/addgene-plasmid-134786-sequence-266694.gbk

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LOCUS sequence-266694- 8100 bp DNA circular SYN 25-AUG-2024 DEFINITION Stable overexpression lentiviral vector of His tagged PSPH. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8100) AUTHORS Park SM, Seo EH, Bae DH, Kim SS, Kim J, Lin W, Kim KH, Park JB, Kim YS, Yin J, Kim SY TITLE Phosphoserine Phosphatase Promotes Lung Cancer Progression through the Dephosphorylation of IRS-1 and a Noncanonical L-Serine-Independent Pathway. JOURNAL Mol Cells. 2019 Aug 31;42(8):604-616. doi: 10.14348/molcells.2019.0160. PUBMED 31446747 REFERENCE 2 (bases 1 to 8100) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.14348/molcells.2019"; journalName: "Mol Cells"; date: "2019-08-31- 31"; volume: "42"; issue: "8"; pages: "604-616" FEATURES Location/Qualifiers source 1..8100 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..227 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 228..408 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 455..580 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1073..1306 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1491..1535 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 1802..1918 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2008..2211 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2162..2182 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2208..2232 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2291..2308 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" promoter 3049..3260 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 3273..3541 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" CDS 3568..4167 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3568..3587) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4064..4084 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 4174..4762 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4227..4247) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4645..4656) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4836..5069 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5141..5262 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5178..5197) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5232..5251 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5302..5437 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5470..5486) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5470..5486) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5483..5505) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5494..5510 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5518..5548) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5563..5584 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5701..5718) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5872..6460) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6631..7491) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7254..7273 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7492..7596) /gene="bla" /label=AmpR promoter primer_bind 7664..7682 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7720..7742) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7842..7861 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8055..8077 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8069..8086 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8070..8086 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg 61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc 121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc 181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt 241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa 421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc 481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac 541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat 601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa 661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga 721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc 781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat 841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa 901 gaccaccgca cagcaagcgg ccactgatct tcagacctgg aggaggagat atgagggaca 961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac 1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt 1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga 1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg 1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg 1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt 1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat 1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt 1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac 1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt 1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag 1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc 1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg 1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa 1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa 1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat 1921 cgatactagt attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 1981 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 2041 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 2101 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 2161 acgcaaatgg gcggtaggcg tgtacggtgg gaggtttata taagcagagc tcgtttagtg 2221 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagattctag 2281 agccaccatg catcatcatc atcatcacgt gaattcgctc gagatggtct cccactcaga 2341 gctgaggaag cttttctact cagcagatgc tgtgtgtttt gatgttgaca gcacggtcat 2401 cagagaagaa ggaatcgatg agctagccaa aatctgtggc gttgaggacg cggtgtcaga 2461 aatgacacgg cgagccatgg gcggggcagt gcctttcaaa gctgctctca cagagcgctt 2521 agccctcatc cagccctcca gggagcaggt gcagagactc atagcagagc aacccccaca 2581 cctgaccccc ggcataaggg agctggtaag tcgcctacag gagcgaaatg ttcaggtttt 2641 cctaatatct ggtggcttta ggagtattgt agagcatgtt gcttcaaagc tcaatatccc 2701 agcaaccaat gtatttgcca ataggctgaa attctacttt aacggtgaat atgcaggttt 2761 tgatgagacg cagccaacag ctgaatctgg tggaaaagga aaagtgatta aacttttaaa 2821 ggaaaaattt cattttaaga aaataatcat gattggagat ggtgccacag atatggaagc 2881 ctgtcctcct gctgatgctt tcattggatt tggaggaaat gtgatcaggc aacaagtcaa 2941 ggataacgcc aaatggtata tcactgattt tgtagagctg ctgggagaac tggaagaata 3001 aggatccgcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca 3061 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga 3121 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg 3181 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg 3241 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc 3301 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct 3361 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg 3421 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc 3481 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat 3541 ccaagctgtg accggcgcct acgctagatg accgagtaca agcccacggt gcgcctcgcc 3601 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc 3661 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa 3721 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc 3781 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc 3841 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc 3901 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg 3961 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc 4021 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac 4081 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg 4141 tgcatgaccc gcaagcccgg tgcctgagtc gacaatcaac ctctggatta caaaatttgt 4201 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct 4261 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat 4321 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg 4381 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag 4441 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc 4501 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg 4561 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc 4621 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc 4681 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc 4741 tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt 4801 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 4861 aagataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 4921 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 4981 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 5041 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 5101 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 5161 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 5221 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 5281 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 5341 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 5401 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agaccaaatt 5461 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 5521 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 5581 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 5641 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 5701 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5761 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5821 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5881 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5941 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6001 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6061 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6121 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6181 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6241 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6301 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6361 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 6421 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 6481 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 6541 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 6601 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 6661 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6721 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 6781 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 6841 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6901 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6961 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7021 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7081 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7141 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7201 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7261 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7321 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7381 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 7441 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 7501 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 7561 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 7621 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag 7681 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 7741 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 7801 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt 7861 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg 7921 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc 7981 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt 8041 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctgacgcg //

mTurquoise2-UtrCH

98824

In vivo visualization of actin cytoskeleton (can be used for colocalization studies)

Dorus Gadella

Chertkova et al bioRxiv 160374

https://media.addgene.org/snapgene-media/v2.0.0/sequences/190903/f3b18c50-00a8-4bfd-8ea4-201ab65d2404/addgene-plasmid-98824-sequence-190903.gbk

full

LOCUS sequence-190903- 5513 bp DNA circular SYN 26-AUG-2024 DEFINITION In vivo visualization of actin cytoskeleton (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5513) AUTHORS Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J TITLE Robust and Bright Genetically Encoded Fluorescent Markers for Highlighting Structures and Compartments in Mammalian Cells JOURNAL bioRxiv 160374 REFERENCE 2 (bases 1 to 5513) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 160374" FEATURES Location/Qualifiers source 1..5513 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 613..1329 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2125..2177 /label=MCS /note="multiple cloning site" polyA_signal 2301..2422 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2338..2357) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2392..2411 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2429..2884) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2566..2587) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2778..2797 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2911..3015 /gene="bla" /label=AmpR promoter promoter 3017..3374 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 3017..3208 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(3040..3060) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3225..3360 /label=SV40 ori /note="SV40 origin of replication" CDS 3409..4203 /codon_start=1 /product="aminoglycoside phosphotransferase" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS 3409..4203 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3463..3482) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4073..4092 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4391..4410) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4435..4482 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4811..5399 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5300..5319 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac 1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggaaccat ggccaagtat ggagaacatg aagccagtcc tgacaatggg 1381 cagaacgaat tcagtgatat cattaagtcc agatctgatg aacacaatga cgtacagaag 1441 aaaaccttta ccaaatggat aaatgctcga ttttcaaaga gtgggaaacc acccatcaat 1501 gatatgttca cagacctcaa agatggaagg aagctattgg atcttctaga aggcctcaca 1561 ggaacatcac tgccaaagga acgtggttcc acaagggtac atgccttaaa taacgtcaac 1621 agagtgctgc aggttttaca tcagaacaat gtggaattag tgaatatagg gggaaccgac 1681 attgtggatg gaaatcacaa actgactttg gggttacttt ggagcatcat tttgcactgg 1741 caggtgaaag atgtcatgaa ggatgtcatg tcggacctgc agcagacgaa cagtgagaag 1801 atcctgctca gctgggtgcg tcagaccacc aggccctaca gccaagtcaa cgtcctcaac 1861 ttcaccacca gctggacaga tggactcgcc tttaatgctg tcctccaccg acataaacct 1921 gatctcttca gctgggataa agttgtcaaa atgtcaccaa ttgagagact tgaacatgcc 1981 ttcagcaagg ctcaaactta tttgggaatt gaaaagctgt tagatcctga agatgttgcc 2041 gttcggcttc ctgacaagaa atccataatt atgtatttaa catctttgtt tgaggtgcta 2101 cctcagcaag tcaccataga ctaactcgag ctcaagcttc gaattctgca gtcgacggta 2161 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg 2221 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 2281 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 2341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 2401 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc 2461 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc 2521 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 2581 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc 2641 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa 2701 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg 2761 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt 2821 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc 2881 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 2941 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 3001 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt 3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3121 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3181 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3241 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3301 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3361 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 3421 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 3481 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 3541 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca 3601 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 3661 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 3721 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 3781 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 3841 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 3901 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc 3961 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 4021 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 4081 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 4141 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 4201 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 4261 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 4321 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg 4381 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 4441 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt 4501 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg 4561 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca 4621 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt 4681 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 4741 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 4801 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 4861 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 4921 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 4981 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 5041 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 5101 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 5161 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 5221 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 5281 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 5341 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 5401 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 5461 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat //

pMV17

128877

Position 2 module- Act1 promoter

Gilles Charvin

Goulev et al PLoS One. 2019 Aug 28;14(8):e0220694. doi: 10.1371/journal.pone.0220694. eCollection 2019.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/252811/5aac3ffd-2f23-4bb8-b764-6bcccfdf4f1c/addgene-plasmid-128877-sequence-252811.gbk

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LOCUS sequence-252811- 3210 bp DNA circular SYN 26-AUG-2024 DEFINITION Position 2 module- Act1 promoter. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3210) AUTHORS Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G TITLE COSPLAY: An expandable toolbox for combinatorial and swift generation of expression plasmids in yeast. JOURNAL PLoS One. 2019 Aug 28;14(8):e0220694. doi: 10.1371/journal.pone.0220694. eCollection 2019. PUBMED 31461461 REFERENCE 2 (bases 1 to 3210) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One."; date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 10.1371/journal.pone.0220694. eCollection 2019" FEATURES Location/Qualifiers source 1..3210 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 4..21 /label=L4440 /note="L4440 vector, forward primer" protein_bind 138..159 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 174..204 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 212..228 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 217..239 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 236..252 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 236..252 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(846..863) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(846..862) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(855..877) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(1071..1090) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1190..1212 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1250..1268) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1336..1440 /gene="bla" /label=AmpR promoter CDS 1441..2301 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1659..1678) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2472..3060 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2961..2980 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 61 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc 121 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac 181 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga 241 aacagctatg accatgatta cgccaagctt gcatgcaggc ctctggcagt cgacgggccc 301 ggtctctgtt ctaatgttaa ttcagtaaat tttcgatctt gggaagaaaa aagcagtaag 361 cgtgaaaaat ctaaaagctg atgtagtaga agatcctatt ctttaacaaa gattgacctt 421 ttctttttct tcttggtttg agtagaaagg ggaaggaaga atacaagaga gaggagaaaa 481 aggaagataa aaagagagcg tgatataaat gaatatatat tgaacaagag agattgggaa 541 ggaaaggatc aaacaaaccc aaaaatattt caaaaaggag agagagaggc gagtttggtt 601 tcaaaacggt ttatttattt atgcaagagg acgtggaaga aaaagaagaa ggaagaaaaa 661 aatttgaaag aaaaaaacgc gtggcgggta aagaagaaaa tggaaaatag aggccgggtg 721 acagagaaat attgagggtt aattagaaag tatgttaggg tgaggcatat gtttttaagg 781 gttttgagca tacgagaccg ggatccgata tctagatgca ttcgcgaggt accgagctcg 841 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt 901 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc 961 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt 1021 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc 1081 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga 1141 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 1201 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata 1261 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 1321 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 1381 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 1441 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 1501 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 1561 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 1621 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 1681 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 1741 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 1801 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 1861 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 1921 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 1981 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 2041 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 2101 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggatct 2161 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 2221 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 2281 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 2341 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 2401 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 2461 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 2521 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2581 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 2641 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2701 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2761 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2821 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2881 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2941 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 3001 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 3061 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 3121 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 3181 gataccgctc gccgcagccg aacgaccgag //

pdCas9_CL

166245

regulated dCas9 generator

Domitilla Del Vecchio

Huang et al Nat Commun. 2021 Mar 16;12(1):1692. doi: 10.1038/s41467-021-21772-6.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/331761/0c38c8f6-ce1b-427c-8c2c-806e82c2916d/addgene-plasmid-166245-sequence-331761.gbk

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LOCUS sequence-331761- 7177 bp DNA circular SYN 26-AUG-2024 DEFINITION regulated dCas9 generator. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7177) AUTHORS Huang HH, Bellato M, Qian Y, Cardenas P, Pasotti L, Magni P, Del Vecchio D TITLE dCas9 regulator to neutralize competition in CRISPRi circuits. JOURNAL Nat Commun. 2021 Mar 16;12(1):1692. doi: 10.1038/s41467-021-21772-6. PUBMED 33727557 REFERENCE 2 (bases 1 to 7177) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun."; date: "2021-03-16"; volume: "12(1):1692. doi"; pages: " 10.1038/s41467-021-21772-6" FEATURES Location/Qualifiers source 1..7177 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 65..83 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_RNA 159..234 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 289..310 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" promoter 329..357 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" RBS 395..403 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS 409..4515 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" terminator 4539..4610 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4626..4653 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_feature 4660..4680 /label=BioBrick suffix /note="universal suffix for all parts" CDS complement(4820..5635) /codon_start=1 /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" CDS complement(4820..5635) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 5543..5562 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 5627..5646 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin complement(6230..6774) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(6309..6328) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 agtctattcc atggtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 61 taaaaatagg cgtatcacga ggcagggcgc gccaaaaaga gtattgactt cgcatctttt 121 tgtacctata atagattcag tattgactat taatcatcgt tttagagcta gaaatagcaa 181 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt 241 tccaattatt gaacacccta acgggtgttt tttttttttt ggtctcccga attcgcggcc 301 gcttctagag ggcgcgccaa aaagagtatt gactattaat catccggctc gtataataga 361 ttcactctag acgagaggaa acgcggtttt aatataagga ggttattaat ggataagaaa 421 tactcaatag gcttagctat cggcacaaat agcgtcggat gggcggtgat cactgatgaa 481 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa 541 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc 601 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag 661 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct 721 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat 781 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat 841 tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt 901 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta 961 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt 1021 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat 1081 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg 1141 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta 1201 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat 1261 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat 1321 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc 1381 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca 1441 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat 1501 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat 1561 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc 1621 tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga 1681 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg 1741 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg 1801 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa 1861 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat 1921 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg 1981 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag 2041 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta 2101 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa 2161 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa 2221 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc 2281 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat 2341 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga 2401 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa 2461 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt 2521 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt 2581 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt 2641 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca 2701 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc 2761 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact 2821 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg 2881 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatgccat tgttccacaa 2941 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt 3001 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga 3061 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa 3121 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact 3181 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat 3241 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct 3301 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc 3361 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt 3421 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag 3481 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac 3541 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa 3601 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc 3661 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga 3721 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa 3781 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta 3841 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta 3901 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct 3961 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt 4021 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat 4081 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag 4141 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat 4201 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat 4261 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa 4321 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt 4381 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat 4441 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag 4501 ctaggaggtg actaactcga gtaaggatct ccaggcatca aataaaacga aaggctcagt 4561 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc 4621 acactggctc accttcgggt gggcctttct gcgtttatat actagtagcg gccgctgcag 4681 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4741 cactcaaagg cggtaatctc gagtcccgtc aagtcagcgt aatgctctgc cagtgttaca 4801 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat 4861 tcatatcagg attatcgata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 4921 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 4981 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 5041 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc 5101 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 5161 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 5221 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 5281 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 5341 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 5401 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 5461 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 5521 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 5581 tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg ctcataacac 5641 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat 5701 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa 5761 cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg 5821 caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg 5881 gctccctcac tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac 5941 cttcttcacg aggcagacct cagcgctagc ggagtgtata ctggcttact atgttggcac 6001 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt 6061 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg 6121 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat 6181 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc 6241 tccgccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac 6301 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc 6361 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc 6421 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg 6481 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac 6541 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag 6601 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc 6661 cagttacctc ggttcaaaga gttggtagct cagagaacct tcaaaaaacc gccctgcaag 6721 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat 6781 catcttatta aggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 6841 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 6901 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 6961 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 7021 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 7081 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 7141 cgcagaagtg gtcctgcaac tttatccgcc tccatcc //

pcDNA3-EGFP-Rac1-T17N

12982

High

Gary Bokoch

Subauste et al J Biol Chem. 2000 Mar 31. 275(13):9725-33.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/212469/8d5a103d-0adc-4c8a-bf66-9010d91bde29/addgene-plasmid-12982-sequence-212469.gbk

full

LOCUS sequence-212469- 6677 bp DNA circular SYN 25-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6677) AUTHORS Subauste MC, Von Herrath M, Benard V, Chamberlain CE, Chuang TH, Chu K, Bokoch GM, Hahn KM TITLE Rho family proteins modulate rapid apoptosis induced by cytotoxic T lymphocytes and Fas. JOURNAL J Biol Chem. 2000 Mar 31. 275(13):9725-33. PUBMED 10734125 REFERENCE 2 (bases 1 to 6677) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Chem. 2000 Mar 31. 275(13):9725-33." FEATURES Location/Qualifiers source 1..6677 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 895..904 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 901..1617 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 901..1617 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(946..967) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1207..1226) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1554..1575 /label=EGFP-C /note="EGFP, forward primer" CDS 1624..2202 /codon_start=1 /product="dominant negative mutant of human Rac family small GTPase 1" /label=Rac1 (T17N) /translation="MQAIKCVVVGDGAVGKNCLLISYTTNAFPGEYIPTVFDNYSANVM VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR GLKTVFDEAIRAVLCPPPVKKRKRKCLLL" promoter complement(2230..2248) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2231..2248) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2268..2285) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2274..2498 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2544..2972 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2631..2650) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2841..2862 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2981..3001) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2986..3315 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3166..3301 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3228..3247 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3382..4176 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3436..3455) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4046..4065 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4350..4471 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4387..4406) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4441..4460 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4520..4536) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4520..4536) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4533..4555) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4544..4560 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4568..4598) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4613..4634 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4751..4768) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4922..5510) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5002..5021) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5681..6541) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6304..6323 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6542..6646) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc 901 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 961 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 1021 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 1081 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 1141 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 1201 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 1261 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 1321 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 1381 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 1441 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 1501 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 1561 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggaa 1621 ttcatgcagg ccatcaagtg tgtggtggtg ggagacggag ctgtaggtaa aaactgccta 1681 ctgatcagtt acacaaccaa tgcatttcct ggagaatata tccctactgt ctttgacaat 1741 tattctgcca atgttatggt agatggaaaa ccggtgaatc tgggcttatg ggatacagct 1801 ggacaagaag attatgacag attacgcccc ctatcctatc cgcaaacaga tgtgttctta 1861 atttgctttt cccttgtgag tcctgcatca tttgaaaatg tccgtgcaaa gtggtatcct 1921 gaggtgcggc accactgtcc caacactccc atcatcctag tgggaactaa acttgatctt 1981 agggatgata aagacacgat cgagaaactg aaggagaaga agctgactcc catcacctat 2041 ccgcagggtc tagccatggc taaggagatt ggtgctgtaa aatacctgga gtgctcggcg 2101 ctcacacagc gaggcctcaa gacagtgttt gacgaagcga tccgagcagt cctctgcccg 2161 cctcccgtga agaagaggaa gagaaaatgc ctgctgttgt aactcgagca tgcatctaga 2221 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc 2281 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 2341 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 2401 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 2461 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 2521 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 2581 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 2641 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 2701 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 2761 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 2821 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 2881 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 2941 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 3001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3061 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3121 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 3181 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 3241 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 3301 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 3361 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 3421 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 3481 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 3541 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 3601 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 3661 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 3721 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 3781 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 3841 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 3901 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 3961 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 4021 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 4081 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 4141 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 4201 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 4261 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 4321 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 4381 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 4441 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 4501 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4561 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4621 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4681 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4741 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4801 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4861 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4921 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5041 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5101 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5161 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5221 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5281 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5341 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 5401 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 5461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5521 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 5581 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 5641 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 5701 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 5761 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5821 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5881 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5941 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6001 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6061 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6121 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6181 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6241 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6301 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6361 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6421 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6481 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6541 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 6601 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 6661 aagtgccacc tgacgtc //

vYM019_PB-Blast-pEF1a-H2B-mCh-D3bCD

197687

plasmid for expressing the DNMT3bCD

Michael Elowitz

Ma et al ACS Synth Biol. 2023 Aug 12. doi: 10.1021/acssynbio.3c00078.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/389531/ff384a24-21df-4297-80da-fb338e4fd2c3/addgene-plasmid-197687-sequence-389531.gbk

full

LOCUS sequence-389531- 10780 bp DNA circular SYN 26-AUG-2024 DEFINITION plasmid for expressing the DNMT3bCD. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10780) AUTHORS Ma Y, Budde MW, Zhu J, Elowitz MB TITLE Tuning Methylation-Dependent Silencing Dynamics by Synthetic Modulation of CpG Density. JOURNAL ACS Synth Biol. 2023 Aug 12. doi: 10.1021/acssynbio.3c00078. PUBMED 37572041 REFERENCE 2 (bases 1 to 10780) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth Biol. 2023 Aug 12. doi: 10.1021/acssynbio.3c00078." FEATURES Location/Qualifiers source 1..10780 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(83..103) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 88..417 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 268..403 /label=SV40 ori /note="SV40 origin of replication" primer_bind 330..349 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 465..512 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 531..929 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" primer_bind complement(989..1009) /label=WPRE-R /note="WPRE, reverse primer" polyA_signal 1051..1172 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" repeat_region 1935..1969 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind complement(2460..2476) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2460..2476) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2473..2495) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2484..2500 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2508..2538) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2553..2574 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2691..2708) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2862..3450) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2942..2961) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3621..4481) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4244..4263 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4482..4586) /gene="bla" /label=AmpR promoter rep_origin complement(4612..5067) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4749..4770) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4961..4980 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5194..5216 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5208..5225 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 5209..5225 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" repeat_region 5355..5417 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" intron 6094..7032 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 6989..7009 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" promoter 7065..7083 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 7144..7521 /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSAK" CDS 7528..8235 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(7673..7691) /label=mCherry-R /note="mCherry, reverse primer" CDS 8239..8292 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" primer_bind complement(10377..10396) /label=T7 /note="T7 promoter, forward primer" promoter complement(10378..10396) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal join(10670..10780,1..11) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" ORIGIN 1 aatgtatctt aaagacacct cgagaccatt accctgttat ccctagaatt cgcggccgca 61 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 241 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 361 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 421 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat 481 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc 541 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc 601 ccatctctga agattacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca 661 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca 721 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca 781 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga 841 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc 901 tgccctctgg ttatgtgtgg gagggctaag tcgacaatca acctctggat tacaaaattt 961 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 1021 ctttaatgcc tttgtatcat gcgttaacta aacttgttta ttgcagctta taatggttac 1081 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 1141 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggaattgact caaatgatgt 1201 caattagtct atcagaagct catctggtct cccttccggg ggacaagaca tccctgttta 1261 atatttaaac agcagtgttc ccaaactggg ttcttatatc ccttgctctg gtcaaccagg 1321 ttgcagggtt tcctgtcctc acaggaacga agtccctaaa gaaacagtgg cagccaggtt 1381 tagccccgga attgactgga ttcctttttt agggcccatt ggtatggctt tttccccgta 1441 tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc 1501 gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg 1561 agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac 1621 gtaattacat ccctgggggc tttggggggg ggctgtccct gatatctata acaagaaaat 1681 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga 1741 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt 1801 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc 1861 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca 1921 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taatctagct 1981 gcatcaggat catatcgtcg ggtctttttt ccggctcagt catcgcccaa gctggcgcta 2041 tctgggcatc ggggaggaag aagcccgtgc cttttcccgc gaggttgaag cggcatggaa 2101 agagtttgcc gaggatgact gctgctgcat tgacgttgag cgaaaacgca cgtttaccat 2161 gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt gaactgttcg ttcaggccac 2221 ctgggatacc agttcgtcgc ggcttttccg gacacagttc cggatggtca gcccgaagcg 2281 catcagcaac ccgaacaata ccggcgacag ccggaactgc cgtgccggtg tgcagattaa 2341 tgacagcggt gcggcgctgg gatattacgt cagcgaggac gggtatcctg gctggatgcc 2401 gcagaaatgg acatggatac cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg 2461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 2521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 2581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 2641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 2701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 2761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 2821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 2881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 2941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 3001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 3061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 3121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 3181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 3241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 3301 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 3361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 3421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 3481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 3541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 3601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 3661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 3721 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 3781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 3841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 3901 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 3961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 4021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 4081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 4141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 4201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 4261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 4321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 4381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 4441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 4501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 4561 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa 4621 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 4681 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 4741 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 4801 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 4861 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 4921 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 4981 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 5041 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg 5101 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 5161 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 5221 ccagtgagcg cgcctcgttc attcacgttt ttgaacccgt ggaggacggg cagactcgcg 5281 gtgcaaatgt gttttacagc gtgatggagc agatgaagat gctcgacacg ctgcagaaca 5341 cgcagctaga ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg 5401 tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa 5461 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt 5521 tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa 5581 caaaactttt atgagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc 5641 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg 5701 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg 5761 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg 5821 aaaaggcctc cacggccaga attctaacta taacggtcct aaggtagcga aggtctcgcg 5881 tactgtgagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga 5941 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc 6001 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt 6061 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg 6121 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag 6181 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc 6241 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc 6301 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc 6361 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat 6421 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc 6481 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc 6541 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt 6601 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga 6661 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag 6721 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca 6781 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg 6841 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag 6901 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc 6961 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt 7021 cttccatttc aggtgtcgtg agctagcgat tacgccaagc tcgaaattaa ccctcactaa 7081 agggaacaaa agctggagct ccaccgcggt ggcggccgct ctagcccggg cggatccgcc 7141 accatgccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa gaaggcggtg 7201 actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga gagctattcc 7261 atctatgtgt acaaggttct gaagcaggtc caccctgaca ccggcatttc gtccaaggcc 7321 atgggcatca tgaattcgtt tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc 7381 cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca gacggccgtg 7441 cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac taaggccatc 7501 accaagtaca ccagcgctaa gcctaggatg gtgagcaagg gcgaggagga taacatggcc 7561 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag 7621 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg 7681 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg 7741 tacggctcca aggcctacgt gaagcacccc gccgacattc ccgactactt gaagctgtcc 7801 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc 7861 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc 7921 accaacttcc cctccgacgg ccccgtaatg cagaagaaaa ccatgggctg ggaggcctcc 7981 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag 8041 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc 8101 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag 8161 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg 8221 gacgagctgt acaagtctga gggcaggggg agcctcctaa cttgcggaga cgtcgaggag 8281 aatcctggcc catctagact ggacaagagc aaagtcataa acggagctct ggaattactc 8341 aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag 8401 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc 8461 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac 8521 tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct 8581 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc 8641 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg 8701 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa 8761 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc 8821 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg 8881 gagaaacagc taaagtgcga aagcggcggg ggtagcggca gcggtagcgc atgcacaacc 8941 aatgactctg ctgcttctga gtccccccca cccaagcgcc tcaagacaaa tagctatggc 9001 gggaaggacc gaggggagga tgaggagagc cgagaacgga tggcttctga agtcaccaac 9061 aacaagggca atctggagga ccgctgtttg tcctgtggaa agaagaaccc tgtgtccttc 9121 caccccctct ttgagggtgg gctctgtcag agttgccggg atcgcttcct agagctcttc 9181 tacatgtatg atgaggacgg ctatcagtcc tactgcaccg tgtgctgtga gggccgtgaa 9241 ctgctgctgt gcagtaacac aagctgctgc agatgcttct gtgtggagtg tctggaggtg 9301 ctggtgggcg caggcacagc tgaggatgcc aagctgcagg aaccctggag ctgctatatg 9361 tgcctccctc agcgctgcca tggggtcctc cgacgcagga aagattggaa catgcgcctg 9421 caagacttct tcactactga tcctgacctg gaagaatttc aggagccacc caagttgtac 9481 ccagcaattc ctgcagccaa aaggaggccc attagagtcc tgtctctgtt tgatggaatt 9541 gcaacggggt acttggtgct caaggagttg ggtattaaag tggaaaagta cattgcctcc 9601 gaagtctgtg cagagtccat cgctgtggga actgttaagc atgaaggcca gatcaaatat 9661 gtcaatgacg tccggaaaat caccaagaaa aatattgaag agtggggccc gttcgacttg 9721 gtgattggtg gaagcccatg caatgatctc tctaacgtca atcctgcccg caaaggttta 9781 tatgagggca caggaaggct cttcttcgag ttttaccact tgctgaatta tacccgcccc 9841 aaggagggcg acaaccgtcc attcttctgg atgttcgaga atgttgtggc catgaaagtg 9901 aatgacaaga aagacatctc aagattcctg gcatgtaacc cagtgatgat cgatgccatc 9961 aaggtgtctg ctgctcacag ggcccggtac ttctggggta acctacccgg aatgaacagg 10021 cccgtgatgg cttcaaagaa tgataagctc gagctgcagg actgcctgga gttcagtagg 10081 acagcaaagt taaagaaagt gcagacaata accaccaagt cgaactccat cagacagggc 10141 aaaaaccagc ttttccctgt agtcatgaat ggcaaggacg acgttttgtg gtgcactgag 10201 ctcgaaagga tcttcggctt ccctgctcac tacacggacg tgtccaacat gggccgcggc 10261 gcccgtcaga agctgctggg caggtcctgg agtgtaccgg tcatcagaca cctgtttgcc 10321 cccttgaagg actactttgc ctgtgaatag ggtaccaggt aagtgtaccc aattcgccct 10381 atagtgagtc gtattacaat tcactcgatc gcccttccca acagttgcgc agcctgaatg 10441 gcgaatggag atccaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg 10501 tgtattttag attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt 10561 catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc 10621 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat 10681 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 10741 tttttcactg cattctagtt gtggtttgtc caaactcatc //

CAG pShuttle

13427

Medium

(Empty Backbone)

Ronald Kahn

pShuttle vectors (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/258367/1ee42f74-bdeb-4e3a-8c25-3b730c0c2b71/addgene-plasmid-13427-sequence-258367.gbk

full

LOCUS sequence-258367- 8965 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8965) TITLE pShuttle vectors REFERENCE 2 (bases 1 to 8965) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..8965 /mol_type="other DNA" /organism="synthetic DNA construct" repeat_region 1..103 /label=ITR /note="inverted terminal repeat of human adenovirus serotype 5" misc_signal 191..341 /label=Ad5 Psi /note="packaging signal for adenovirus serotype 5" enhancer 411..790 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 1050..2066 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1990..2012 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2074..2093 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" primer_bind complement(2259..2278) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2324..2379 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2378..2397) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" repeat_region 5924..6026 /label=ITR /note="inverted terminal repeat of human adenovirus serotype 5" primer_bind complement(6115..6132) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6286..6874) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6366..6385) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 7706..8500 /codon_start=1 /product="aminoglycoside phosphotransferase" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS 7706..8500 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7760..7779) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 8370..8389 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin complement(8548..8928) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8610..8631) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8822..8841 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt 61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt 121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg 181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag 241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga 301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tactggtacc gggccccccc 361 tcgacattga ttattgacta gttattaata gtaatcaatt acsscccctc gacattgatt 421 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 481 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 541 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 601 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 661 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 721 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 781 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc 841 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg 901 gggggggggg gggggcgcgc gggggggggg gsggggcgrg gcggagaggt gcggcggcag 961 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc 1021 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc 1081 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac 1141 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac 1201 ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt 1261 gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg 1321 gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc 1381 gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg 1441 aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg 1501 cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg 1561 cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt 1621 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg 1681 aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt 1741 gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg 1801 agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg 1861 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc 1921 ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg 1981 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg 2041 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc 2101 atcattttgg caaagaatta gcggccgcgc tactatttaa atgatatcaa gcttcctgca 2161 gggtcgacgg taccaggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct 2221 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat 2281 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt 2341 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg 2401 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat 2461 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc 2521 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata 2581 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta 2641 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg 2701 agatccctcg acccctcgag ggcccaagct gtttggggga tctgggcgtg gttaagggtg 2761 ggaaagaata tataaggtgg gggtcttatg tagttttgta tctgttttgc agcagccgcc 2821 gccgccatga gcaccaactc gtttgatgga agcattgtga gctcatattt gacaacgcgc 2881 atgcccccat gggccggggt gcgtcagaat gtgatgggct ccagcattga tggtcgcccc 2941 gtcctgcccg caaactctac taccttgacc tacgagaccg tgtctggaac gccgttggag 3001 actgcagcct ccgccgccgc ttcagccgct gcagccaccg cccgcgggat tgtgactgac 3061 tttgctttcc tgagcccgct tgcaagcagt gcagcttccc gttcatccgc ccgcgatgac 3121 aagttgacgg ctcttttggc acaattggat tctttgaccc gggaacttaa tgtcgtttct 3181 cagcagctgt tggatctgcg ccagcaggtt tctgccctga aggcttcctc ccctcccaat 3241 gcggtttaaa acataaataa aaaaccagac tctgtttgga tttggatcaa gcaagtgtct 3301 tgctgtcttt atttaggggt tttgcgcgcg cggtaggccc gggaccagcg gtctcggtcg 3361 ttgagggtcc tgtgtatttt ttccaggacg tggtaaaggt gactctggat gttcagatac 3421 atgggcataa gcccgtctct ggggtggagg tagcaccact gcagagcttc atgctgcggg 3481 gtggtgttgt agatgatcca gtcgtagcag gagcgctggg cgtggtgcct aaaaatgtct 3541 ttcagtagca agctgattgc caggggcagg cccttggtgt aagtgtttac aaagcggtta 3601 agctgggatg ggtgcatacg tggggatatg agatgcatct tggactgtat ttttaggttg 3661 gctatgttcc cagccatatc cctccgggga ttcatgttgt gcagaaccac cagcacagtg 3721 tatccggtgc acttgggaaa tttgtcatgt agcttagaag gaaatgcgtg gaagaacttg 3781 gagacgccct tgtgacctcc aagattttcc atgcattcgt ccataatgat ggcaatgggc 3841 ccacgggcgg cggcctgggc gaagatattt ctgggatcac taacgtcata gttgtgttcc 3901 aggatgagat cgtcataggc catttttaca aagcgcgggc ggagggtgcc agactgcggt 3961 ataatggttc catccggccc aggggcgtag ttaccctcac agatttgcat ttcccacgct 4021 ttgagttcag atggggggat catgtctacc tgcggggcga tgaagaaaac ggtttccggg 4081 gtaggggaga tcagctggga agaaagcagg ttcctgagca gctgcgactt accgcagccg 4141 gtgggcccgt aaatcacacc tattaccggc tgcaactggt agttaagaga gctgcagctg 4201 ccgtcatccc tgagcagggg ggccacttcg ttaagcatgt ccctgactcg catgttttcc 4261 ctgaccaaat ccgccagaag gcgctcgccg cccagcgata gcagttcttg caaggaagca 4321 aagtttttca acggtttgag accgtccgcc gtaggcatgc ttttgagcgt ttgaccaagc 4381 agttccaggc ggtcccacag ctcggtcacc tgctctacgg catctcgatc cagcatatct 4441 cctcgtttcg cgggttgggg cggctttcgc tgtacggcag tagtcggtgc tcgtccagac 4501 gggccagggt catgtctttc cacgggcgca gggtcctcgt cagcgtagtc tgggtcacgg 4561 tgaaggggtg cgctccgggc tgcgcgctgg ccagggtgcg cttgaggctg gtcctgctgg 4621 tgctgaagcg ctgccggtct tcgccctgcg cgtcggccag gtagcatttg accatggtgt 4681 catagtccag cccctccgcg gcgtggccct tggcgcgcag cttgcccttg gaggaggcgc 4741 cgcacgaggg gcagtgcaga cttttgaggg cgtagagctt gggcgcgaga aataccgatt 4801 ccggggagta ggcatccgcg ccgcaggccc cgcagacggt ctcgcattcc acgagccagg 4861 tgagctctgg ccgttcgggg tcaaaaacca ggtttccccc atgctttttg atgcgtttct 4921 tacctctggt ttccatgagc cggtgtccac gctcggtgac gaaaaggctg tccgtgtccc 4981 cgtatacaga cttgagaggg agtttaaacg aattcaatag cttgttgcat gggcggcgat 5041 ataaaatgca aggtgctgct caaaaaatca ggcaaagcct cgcgcaaaaa agaaagcaca 5101 tcgtagtcat gctcatgcag ataaaggcag gtaagctccg gaaccaccac agaaaaagac 5161 accatttttc tctcaaacat gtctgcgggt ttctgcataa acacaaaata aaataacaaa 5221 aaaacattta aacattagaa gcctgtctta caacaggaaa aacaaccctt ataagcataa 5281 gacggactac ggccatgccg gcgtgaccgt aaaaaaactg gtcaccgtga ttaaaaagca 5341 ccaccgacag ctcctcggtc atgtccggag tcataatgta agactcggta aacacatcag 5401 gttgattcac atcggtcagt gctaaaaagc gaccgaaata gcccggggga atacataccc 5461 gcaggcgtag agacaacatt acagccccca taggaggtat aacaaaatta ataggagaga 5521 aaaacacata aacacctgaa aaaccctcct gcctaggcaa aatagcaccc tcccgctcca 5581 gaacaacata cagcgcttcc acagcggcag ccataacagt cagccttacc agtaaaaaag 5641 aaaacctatt aaaaaaacac cactcgacac ggcaccagct caatcagtca cagtgtaaaa 5701 aagggccaag tgcagagcga gtatatatag gactaaaaaa tgacgtaacg gttaaagtcc 5761 acaaaaaaca cccagaaaac cgcacgcgaa cctacgccca gaaacgaaag ccaaaaaacc 5821 cacaacttcc tcaaatcgtc acttccgttt tcccacgtta cgtcacttcc cattttaaga 5881 aaactacaat tcccaacaca tacaagttac tccgccctaa aacctacgtc acccgccccg 5941 ttcccacgcc ccgcgccacg tcacaaactc caccccctca ttatcatatt ggcttcaatc 6001 caaaataagg tatattattg atgatgttaa ttaacatgca tggatccata tgcggtgtga 6061 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct 6121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 6181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 6241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 6301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 6361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 6421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 6481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 6541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 6601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 6661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 6721 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6961 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 7021 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 7081 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 7141 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 7201 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 7261 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 7321 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcagccatga gattatcaaa 7381 aaggatcttc acctagatcc ttttcacgta gaaagccagt ccgcagaaac ggtgctgacc 7441 ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa 7501 gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc 7561 aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt 7621 aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gctctgatca 7681 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 7741 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 7801 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 7861 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac 7921 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 7981 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 8041 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 8101 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 8161 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 8221 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 8281 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 8341 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 8401 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 8461 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attttgttaa aatttttgtt 8521 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 8581 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 8641 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 8701 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 8761 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 8821 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc 8881 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt 8941 caggatcgaa ttaattctta attaa //

pMuLE ENTR CMV-hCas9 R4-R3

62132

MuLE (Multiple Lentiviral Expression) Entry vector containing CMV promoter and human codon optimized Cas9 module Compatible with MultiSite Gateway cloning

Ian Frew

Albers et al J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/101416/28232def-2b2a-4823-93f3-119d51a7a64b/addgene-plasmid-62132-sequence-101416.gbk

full

LOCUS sequence-101416- 7523 bp DNA circular SYN 26-AUG-2024 DEFINITION MuLE (Multiple Lentiviral Expression) Entry vector containing CMV promoter and human codon optimized Cas9 module Compatible with MultiSite Gateway cloning. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7523) AUTHORS Albers J, Danzer C, Rechsteiner M, Lehmann H, Brandt LP, Hejhal T, Catalano A, Busenhart P, Goncalves AF, Brandt S, Bode PK, Bode-Lesniewska B, Wild PJ, Frew IJ TITLE A versatile modular vector system for rapid combinatorial mammalian genetics. JOURNAL J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743. PUBMED 25751063 REFERENCE 2 (bases 1 to 7523) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743." FEATURES Location/Qualifiers source 1..7523 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 9..132 /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter complement(188..206) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(211..227) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(211..227) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(329..348) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 340..1149 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(407..426) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 1296..1884 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1785..1804 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2038..2055 /label=L4440 /note="L4440 vector, forward primer" terminator complement(2214..2241) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(2333..2419) /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 2468..2490 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2482..2499 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2483..2499 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind complement(2549..2673) /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" enhancer 2700..3079 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3080..3283 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3234..3254 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 3328..3346 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 3366..3375 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 3372..7475 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 3372..7475 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 3372..7475 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 7488..7508 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" ORIGIN 1 ctagaggaca actttgtata ataaagttga acgagaaacg taaaatgata taaatatcaa 61 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat 121 ccagtcacta tgaatcaact acttagatgg tattagtgac ctgtactgca gggcggccgc 181 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc 241 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata 301 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa 361 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct 421 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg 481 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg 541 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt 601 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta 661 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc 721 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc 781 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag 841 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca 901 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg 961 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt 1021 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa 1081 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag 1141 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga 1201 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga 1261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 1321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 1381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 1441 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 1501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 1561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 1621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 1681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 1741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 1801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 1861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 1921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 1981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc 2041 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt 2101 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag 2161 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc 2221 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg 2281 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc 2341 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact 2401 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg 2461 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gcccctacag 2521 gtcactaata ccatctaagt agttgattca tagtgactgg atatgttgtg ttttacagta 2581 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta 2641 cgtttctcgt tcaacttttc tatacaaagt tggcgatgta cgggccagat atacgcgttg 2701 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc 2761 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 2821 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 2881 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 2941 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 3001 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 3061 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 3121 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 3181 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 3241 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga 3301 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg 3361 gtaccgccac catggacaag aagtactcca ttgggctcga tatcggcaca aacagcgtcg 3421 gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca 3481 ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg 3541 agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga 3601 atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt 3661 tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac gagcgccacc 3721 caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc 3781 atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg 3841 cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag 3901 acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat cagcttttcg 3961 aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt 4021 ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc 4081 tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg 4141 acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca 4201 atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt 4261 cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc 4321 tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg 4381 cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa 4441 atggctacgc cggatacatt gacggcggag caagccagga ggaattttac aaatttatta 4501 agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag 4561 atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg 4621 gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca 4681 gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc 4741 ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga 4801 acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta 4861 actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt 4921 acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc 4981 cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc 5041 ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa tgtttcgact 5101 ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc 5161 tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg 5221 aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga 5281 aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata 5341 caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag cagagtggaa 5401 agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc atgcagttga 5461 tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg 5521 gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa 5581 tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg 5641 agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca 5701 gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta 5761 aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc 5821 agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg 5881 acgtggatca tatcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt 5941 tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa gaagttgtca 6001 agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt 6061 tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca 6121 tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt 6181 cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta 6241 ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag 6301 agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac 6361 ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg 6421 atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact 6481 tcttttacag caatattatg aattttttca agaccgagat tacactggcc aatggagaga 6541 ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg 6601 gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa 6661 agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg 6721 acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc 6781 ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac 6841 tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa 6901 accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta 6961 agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg 7021 cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt 7081 atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc 7141 tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct 7201 ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc 7261 acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt actctgacca 7321 acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca 7381 cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg 7441 aaacaagaat cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga 7501 ggaaggtgtg agcggccgcg act //

pHR_EF1a-->myc-mtLaG2

162224

Lentiviral vector for constitutive expression of Myc GFP nanobody Lag2 in mammalian cells

Wendell Lim

Toda et al Science. 2020 Oct 16;370(6514):327-331. doi: 10.1126/science.abc0033.

partial

pNb4OMT-A53M

216232

Expresses a mutant Norbelladine 4'-O-Methyltransferase

d'Oelsnitz et al Nat Commun. 2024 Mar 7;15(1):2084. doi: 10.1038/s41467-024-46356-y.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/437622/8e22cbe4-c549-46b3-84bd-bfa47bf82a43/addgene-plasmid-216232-sequence-437622.gbk

full

LOCUS sequence-437622- 3986 bp DNA circular SYN 25-AUG-2024 DEFINITION Expresses a mutant Norbelladine 4'-O-Methyltransferase. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3986) AUTHORS d'Oelsnitz S, Diaz DJ, Kim W, Acosta DJ, Dangerfield TL, Schechter MW, Minus MB, Howard JR, Do H, Loy JM, Alper HS, Zhang YJ, Ellington AD TITLE Biosensor and machine learning-aided engineering of an amaryllidaceae enzyme. JOURNAL Nat Commun. 2024 Mar 7;15(1):2084. doi: 10.1038/s41467-024-46356-y. PUBMED 38453941 REFERENCE 2 (bases 1 to 3986) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun."; date: "2024-03-7"; volume: "15(1):2084. doi"; pages: " 10.1038/s41467-024-46356-y" FEATURES Location/Qualifiers source 1..3986 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 24..215 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 372..391 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(466..483) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(637..1225) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(717..736) /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_RNA 1494..1543 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" protein_bind complement(2482..2515) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" promoter 2526..2597 /gene="bla" /label=AmpR promoter CDS 2598..3458 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2816..2835) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" protein_bind complement(3592..3625) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" ORIGIN 1 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc 61 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gatgcggatg 121 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc 181 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg cagacggtca 241 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 301 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg 361 gcttaactat gcggcatcag agcagattgt actgagagtg caccggtgtg aaataccgca 421 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc 481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 661 cgagcatcac aaaaattgac gctcaagtca gaggtggcga aacccgacag gactataaag 721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac 1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 1261 tcagtggaac gaaaactcac gttaaggccc tctccaagac cgagccatca acaaagcgtc 1321 tcgctgaggt ttcatggagc ctctggttca tctccggcaa ttaaaaaagc ggctaaccac 1381 gccgcttttt ttacgtctgc aggaacgggc tgtcgaccgg cgtgccgttc gtttaccgct 1441 agctcagtcc taggtacaat tacagccatc gtacgagccc tggctgagca cagctgtcac 1501 cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac aaataatttt 1561 gtttaaacta gtgaaccacg aggcctacat atgggtgctt caattgatga ctactccttg 1621 gtacataaaa acatcttgca ttccgaagat ctgctgaaat acattcttga aaccagtgca 1681 tatcctcgcg aacacgaaca attaaagggt ctgcgtgagg ttactgaaaa gcacgaatgg 1741 tcatccatgt tagtaccggc agacgaaggt ttattcctgt caatgttact gaagttgatg 1801 aatgcaaaac gtactatcga aatcggcgtg tacacgggat acagcctgtt aacaacggca 1861 ttggctttac cggaggatgg taaaattacg gcgatcgatg taaataagag ttattacgaa 1921 atcggattgc cctttattca gaaggcgggc gtggaacaca agatcaactt tattgagtcg 1981 gaagcgcttc ccgtgctgga tcaaatgtta gaggagatga aggaggagga tttatacgac 2041 tacgcttttg tggatgctga taaaagcaat tatgccaatt accatgagcg tcttgtaaaa 2101 cttgtacgta tcggtggtgc catcctgtac gacaatacac tgtggtatgg ttctgttgcg 2161 tacccggaat accccggtct gcatccagag gaagaagtcg cgcgtctgag ctttcgtaac 2221 ttgaatacct ttttagcagc agatcctcgc gtagaaatta gtcaagtctc aattggtgat 2281 ggcgtgacca tttgtcgccg cttgtattaa taatcctatc gccactttca gccaaaaaac 2341 ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg gtggacagga tcggcggttt 2401 tcttttctct tctcaacacc cttcgcgtca acacttttcc gccaaggaga cggttggtca 2461 ggttttcggg aggtgtggct ggaagttcct atactttcta gagaatagga acttctttct 2521 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 2581 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 2641 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 2701 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 2761 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 2821 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact 2881 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 2941 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 3001 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 3061 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 3121 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg 3181 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 3241 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 3301 ccggtgagcg tggatcgcgc ggtatcattg cagcactggg gccagatggt aagccctccc 3361 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 3421 tcgctgagat aggtgcctca ctgattaagc attggtaagt tgtgatggcg gtaggaatgt 3481 aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt 3541 ttagggaaag agcatttgtc atcccgttga atatggctcc cttaacgtga ggaagttcct 3601 atactttcta gagaatagga acttctacag atggacttgg gttggcggtt tcaggagtct 3661 gcaaaacgct gcgacctgag caacaacatg aatggtcatc ggtttccgtg tttcgtaaag 3721 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg 3781 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg 3841 agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc 3901 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat 3961 cggtatcatt acccccatga acagaa //

ADK gRNA (BRDN0001146463)

75742

3rd generation lentiviral gRNA plasmid targeting human ADK

Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/184478/03c4e372-033f-4890-aaf2-aa7e44b2caad/addgene-plasmid-75742-sequence-184478.gbk

full

LOCUS sequence-184478- 8323 bp DNA circular SYN 26-AUG-2024 DEFINITION 3rd generation lentiviral gRNA plasmid targeting human ADK. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8323) AUTHORS Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE TITLE Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. JOURNAL Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. PUBMED 26780180 REFERENCE 2 (bases 1 to 8323) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437." FEATURES Location/Qualifiers source 1..8323 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 87..1265 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 318..1256 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1213..1233 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1278..1877 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1278..1297) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1774..1794 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 1893..2481 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1946..1966) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2364..2375) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 2553..2786 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2864..2985 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2901..2920) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2955..2974 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3025..3160 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(3180..3199) /label=T7 /note="T7 promoter, forward primer" promoter complement(3181..3199) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3209..3226) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3209..3225) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3218..3240) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3367..3822 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3454..3473) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3664..3685 /label=F1ori-F /note="F1 origin, forward primer" promoter 3848..3952 /gene="bla" /label=AmpR promoter CDS 3953..4813 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(4171..4190) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4984..5572 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5473..5492 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5726..5743 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5860..5881 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5896..5926 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5934..5950 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 5939..5961 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5958..5974 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5958..5974 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5993..6013 /label=T3 /note="T3 promoter, forward primer" promoter 5995..6013 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6041..6267 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6268..6448 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6495..6620 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7113..7346 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7531..7575 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 7753..8001 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)" primer_bind 7753..7773 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 7924..7943 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 8023..8098 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 8154..8271 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct 61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat 121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa 181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg 241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt 301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg 361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc 421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt 481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt 541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat 601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc 661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca 721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc 781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg 841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc 901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac 961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt 1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag 1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag 1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat 1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt 1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg 1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc 1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca 1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg 1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca 1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc 1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc 1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg 1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg 1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc 1861 gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg 1921 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 1981 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 2041 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 2101 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 2161 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 2221 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 2281 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg 2341 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 2401 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 2461 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga 2521 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag 2581 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga 2641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 2701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 2761 ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca 2821 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 2881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 2941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 3001 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 3121 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc 3181 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg 3241 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc 3301 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg 3361 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 3421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 3481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 3541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 3601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 3661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 3721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 3781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg 3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 3961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 4021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 4081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 4141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 4201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 4261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 4321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 4381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 4441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 4501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 4561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 4621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 4681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 4741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 4801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 5281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 5701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 5761 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 5821 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca 5881 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag 5941 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa 6001 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc 6061 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag 6121 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa 6181 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta 6241 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg 6301 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 6361 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 6421 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag 6481 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag 6541 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga 6601 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa 6661 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca 6721 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac 6781 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat 6841 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg 6901 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg 6961 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat 7021 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga 7081 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga 7141 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta 7201 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat 7261 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa 7321 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc 7381 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat 7441 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc 7501 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat 7561 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta 7621 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata 7681 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg 7741 aggggaccca gagagggcct atttcccatg attccttcat atttgcatat acgatacaag 7801 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat 7861 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa 7921 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc 7981 ttgtggaaag gacgaaacac cgtctggaga aaaactggat gtgttttaga gctagaaata 8041 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt 8101 ttttaagctt ggcgtaacta gatcttgaga caaatggcag tattcatcca caattttaaa 8161 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca 8221 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat 8281 tacagggaca gcagagatcc actttggcgc cggctcgagg ggg //

pEB1-mYPet

103988

Plasmid encoding mYPet

Philippe Cluzel

Balleza et al Nat Methods. 2018 Jan;15(1):47-51. doi: 10.1038/nmeth.4509. Epub 2017 Nov 20.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/200409/befb4895-1e61-42ea-bc11-22a2e4976201/addgene-plasmid-103988-sequence-200409.gbk

full

LOCUS sequence-200409- 4391 bp DNA circular SYN 26-AUG-2024 DEFINITION Plasmid encoding mYPet. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4391) AUTHORS Balleza E, Kim JM, Cluzel P TITLE Systematic characterization of maturation time of fluorescent proteins in living cells. JOURNAL Nat Methods. 2018 Jan;15(1):47-51. doi: 10.1038/nmeth.4509. Epub 2017 Nov 20. PUBMED 29320486 REFERENCE 2 (bases 1 to 4391) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth.4509"; journalName: "Nat Methods"; date: "2018-01"; volume: "15"; issue: "1"; pages: "47-51" FEATURES Location/Qualifiers source 1..4391 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 61..79 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(294..322) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" terminator 1018..1104 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(1642..2592) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 2640..2862 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator complement(3354..3448) /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3479..4273) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3590..3609) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4200..4219 /label=Neo-R /note="Neomycin resistance gene, reverse primer" ORIGIN 1 gccccagctg gcaattccga cgtctaagaa accattatta tcatgacatt aacctataaa 61 aataggcgta tcacgaggcc ctttcgtctt caccacagct aacaccacgt cgtccctatc 121 tgctgcccta ggtctatgag tggttgctgg ataactttac gggcatgcat aaggctcgta 181 tgatatattc agggagacca caacggtttc cctctacaaa taattttgtt taacttttct 241 agatttaaga aggagatata catatgagta aaggagaaga acttttcact ggagttgtcc 301 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg 361 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt gttatgcact actggaaaac 421 tacctgttcc atggccaaca cttgtcacta ctttgggtta tggtgttcaa tgctttgcga 481 gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg 541 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca 601 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag 661 atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca 721 cggcagacaa acaaaagaat ggaatcaaag cgaacttcaa aattagacac aacattgaag 781 atggaggtgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg 841 tccttttacc agacaaccat tacctgtcct accaatctaa gcttttcaaa gatcccaacg 901 aaaagagaga ccacatggtc cttcttgagt ttttaacagc tgctgggatt acagaaggca 961 tgaatgaact atacaaataa atgtccagac ctgcaggcat gcaagctcta gaggcatcaa 1021 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg 1081 aacgctctcc tgagtaggac aaatccgccg ccctagacct agggtacggg ttttgctgcc 1141 cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc cggcttcagc 1201 cggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt ccccacggga 1261 ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat cgcctgtttc 1321 aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc aatttcatgt 1381 tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg ctgttcatct 1441 gttacattgt cgatctgttc atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc 1501 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt 1561 gatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata 1621 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt 1681 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact tactttgcat 1741 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg 1801 tagtgttttt cttagtccgt tatgtaggta ggaatctgat gtaatggttg ttggtatttt 1861 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc 1921 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt 1981 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct 2041 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat 2101 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat 2161 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt 2221 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct 2281 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct 2341 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt 2401 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc 2461 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt 2521 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac 2581 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg 2641 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg 2701 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc 2761 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata 2821 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc 2881 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa 2941 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt 3001 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca 3061 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa 3121 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg 3181 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc 3241 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt 3301 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt ccctagtgct tggattctca 3361 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag 3421 gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctc 3481 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac 3541 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg 3601 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc 3661 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga 3721 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga 3781 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac 3841 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg 3901 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag 3961 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag 4021 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg 4081 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg 4141 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg 4201 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat 4261 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc 4321 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct 4381 taccagaggg c //

T4 Lysozyme T157C

18638

Brian Matthews

T4 Lysozyme Plasmids - Tray 5 (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/24423/e4f12e93-e198-4e81-98ac-56c39ef5fb72/addgene-plasmid-18638-sequence-24423.gbk

partial

LOCUS sequence-24423-e 790 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 790) TITLE T4 Lysozyme Plasmids - Tray 5 REFERENCE 2 (bases 1 to 790) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..790 /mol_type="genomic DNA" /organism="unspecified" primer_bind complement(711..733) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 722..738 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(746..773) /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" ORIGIN 1 antggaaaag cccgctcatt aggcgggctc agatctgctc atgtttgaca gcttatcatc 61 gataagcttc ggcaataaat tcttgatatg ttttcatatg tttcctttaa atattttaat 121 tagtaattgt ctattcaagt aattgtgaat atactatcac aattccaaga gaaagtaaac 181 agctttatag atttttatac gcgtcccaac agccagttct aaacgttgta atgactcgtt 241 ttgcgcgatt aggtgtttga ttataccatc tacttttagc taagttaact gctgcttcat 301 cccagcgttt ttgttgaagc atacgtaaag agttagtaaa tcctgccaca ccggtttctc 361 ccatttggaa aaccatatta atcaatgcac agcgacgaac cgcatcaaga gaatcataaa 421 ccggttttaa tttagcattt ctcagaattc cgcgaacagc agcatcaaca tcctgattaa 481 agagtttttc agcctcatct tttgtaatta caccattgca attacgccca atagctttat 541 ctaattcaga tttagcagca ttaagtgatg gactttttgt aagcaaatga ccgatgccaa 601 tagtgtaata gccttctgtg tctttataga ttttaagtct aagaccttca tctatacgta 661 acatttcaaa tatattcata atacctccta agcatcgatg gatcctgttt cctgtgtgaa 721 attgttatcc gctcacaatt ccacacatta tacgagccga tgatnaattg tcacaggggg 781 atggggagta //

III-CGCC

43576

Ding et al Cell Stem Cell. 2012 Dec 12. pii: S1934-5909(12)00645-5. doi: 10.1016/j.stem.2012.11.011.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/61572/cfd0834c-57fc-4603-b579-ffb9699944e1/addgene-plasmid-43576-sequence-61572.gbk

partial

LOCUS sequence-61572-e 861 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 861) AUTHORS Ding Q, Lee YK, Schaefer EA, Peters DT, Veres A, Kim K, Kuperwasser N, Motola DL, Meissner TB, Hendriks WT, Trevisan M, Gupta RM, Moisan A, Banks E, Friesen M, Schinzel RT, Xia F, Tang A, Xia Y, Figueroa E, Wann A, Ahfeldt T, Daheron L, Zhang F, Rubin LL, Peng LF, Chung RT, Musunuru K, Cowan CA TITLE A TALEN Genome-Editing System for Generating Human Stem Cell-Based Disease Models. JOURNAL Cell Stem Cell. 2012 Dec 12. pii: S1934-5909(12)00645-5. doi: 10.1016/j.stem.2012.11.011. PUBMED 23246482 REFERENCE 2 (bases 1 to 861) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem Cell. 2012 Dec 12. pii: S1934-5909(12)00645-5. doi: 10.1016/j.stem.2012.11.011." FEATURES Location/Qualifiers source 1..861 /mol_type="genomic DNA" /organism="unspecified" promoter complement(600..618) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(601..618) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(636..652) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(636..652) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(649..671) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 660..676 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(684..714) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 729..750 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 gaattgggcc ctctagatgc atgctcgagc ggccgccagt gtgatggata tctgcagaat 61 tcgcccttgc cgtcctagcg cgtctcggct caccccagag caggtcgtgg cgatcgcaag 121 ccacgacgga ggaaagcaag ccttggaaac agtacagagg ctgttgcctg tgctgtgcca 181 agcgcacgga ctcaccccag agcaggtcgt ggcaatcgcg agcaataacg gcggaaaaca 241 ggctttggaa acggtgcaga ggctccttcc agtgctgtgc caagcgcacg gcctcacccc 301 agagcaggtc gtggcgatcg caagccacga cggaggaaag caagccttgg aaacagtaca 361 gaggctgttg cctgtgctgt gccaagcgca cggattaacc ccagagcagg tcgtggcgat 421 cgcaagccac gacggaggaa agcaagcctt ggaaacagta cagaggctgt tgcctgtgct 481 gtgccaagcg cacggactca gagacgtggg cacaggaaag aagggcgaat tccagcacac 541 tggcggccgt tactagtgga tccgagctcg gtaccaagct tgatgcatag cttgagtatt 601 ctatagtgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 661 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 721 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 781 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 841 ttgcgtattg ggcgctcttc c //

pGC309

19652

Jane Hubbard

Voutev et al Genetics. 2008 Sep . 180(1):103-19.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/9284/fa7f25fe-5d1c-4765-adba-3e474c80aeee/addgene-plasmid-19652-sequence-9284.gbk

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LOCUS sequence-9284-ex 4707 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4707) AUTHORS Voutev R, Hubbard EJ TITLE A ""FLP-Out"" system for controlled gene expression in Caenorhabditis elegans. JOURNAL Genetics. 2008 Sep . 180(1):103-19. PUBMED 18723890 REFERENCE 2 (bases 1 to 4707) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genetics. 2008 Sep . 180(1):103-19." FEATURES Location/Qualifiers source 1..4707 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind complement(78..125) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 202..222 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 244..921 /codon_start=1 /product="rapidly maturing tetrameric variant of DsRed fluorescent protein (Bevis and Glick, 2002)" /label=DsRed-Express /note="mammalian codon-optimized" /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFL" CDS 964..1638 /codon_start=1 /product="rapidly maturing tetrameric variant of DsRed fluorescent protein (Bevis and Glick, 2002)" /label=DsRed-Express /note="mammalian codon-optimized" /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL FL" primer_bind complement(2147..2169) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2269..2288 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(2366..2384) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2452..2556 /gene="bla" /label=AmpR promoter CDS 2557..3417 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2775..2794) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3588..4176 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4077..4096 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4330..4347 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4464..4485 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4500..4530 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4538..4554 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 4543..4565 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4562..4578 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4562..4578 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cactagttct 61 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga 121 acttcaagat ccccggggta tgtttcgaat gatactaaca taacatagaa cattttcagg 181 agctcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggtaga cgataaggat 241 ccgatggtgg cctcctccga ggacgtcatc aaagagttca tgcgcttcaa ggtgcgcatg 301 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac 361 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg 421 gacatcctgt ccccccagtt ccagtacggc tccaaggcgt acgtgaagca ccccgccgac 481 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac 541 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcacgctg 601 atctacaagg tgaagttccg cggcaccaac ttcccccccg acggccccgt aatgcagaag 661 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag 721 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag 781 accatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacaccaag 841 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag 901 ggccgccacc acctgttcct ggggcatggc accggcagca ccggcagcgg cagctccggc 961 accgcctcct ccgaggacgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc 1021 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc 1081 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc 1141 ctgtcccccc agttccagta cggctccaag gcgtacgtga agcaccccgc cgacatcccc 1201 gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag 1261 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac 1321 aaggtgaagt tccgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc 1381 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag 1441 atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc 1501 tacatggcca agaagcccgt gcagctgccc ggctactact acgtggacac caagctggac 1561 atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc 1621 caccacctgt tcctgtagga attcgctagc cggccataca agtaatccgg atgatcgacg 1681 ccaacgtcgt tgaattttca aattttaaat actgaatatt tgtttttttt cctattattt 1741 atttattctc tttgtgtttt ttttcttgct ttctaaaaaa ttaattcaat ccaaatctaa 1801 acatgagcgg ttttttttct ctttccgtct cccaattcgt attccgctcc tctcatctga 1861 acacaatgtg caagtttatt tatcttctcg ctttcatttc attaggacgt ggggggaatt 1921 ggtggaaggg ggaaacacac aaaaggatga tggaaatgaa ataaggacac acaatatgca 1981 acaacattca attcagaaat atggaggaag gtttaaaaga aaacataaaa atatatagag 2041 gaggaaggaa aactagtaaa aaataagcaa agaaattagg cgaacgatga gaattgtcct 2101 cgcttgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 2161 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 2221 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 2281 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 2341 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc 2401 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 2461 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 2521 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 2581 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 2641 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 2701 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 2761 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 2821 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 2881 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 2941 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 3001 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 3061 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 3121 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 3181 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 3241 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 3301 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 3361 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 3421 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 3481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 3541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 3601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 3661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 3721 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 3781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 3841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 3901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 3961 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac 4021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 4081 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 4141 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 4201 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 4261 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 4321 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct 4381 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa 4441 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct 4501 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac 4561 acaggaaaca gctatgacca tgattacgcc aagctgtaag tttaaacatg atcttactaa 4621 ctaactattc tcatttaaat tttcagagct taaaaatggc tgaaatcact cacaacgatg 4681 gatacgctaa caacttggaa atgaaat //

pUC19-Chantico

170659

pUC19 vector containing an insertion of the Chantico promoter

Alexander Ruthenburg

Hall et al bioRxiv 2021.12.23.474040

https://media.addgene.org/snapgene-media/v2.0.0/sequences/335913/689bd3f6-cdd2-4f63-95d5-e06ae2785b70/addgene-plasmid-170659-sequence-335913.gbk

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LOCUS sequence-335913- 3811 bp DNA circular SYN 26-AUG-2024 DEFINITION pUC19 vector containing an insertion of the Chantico promoter. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3811) AUTHORS Hall JM, Kainth AS, Rowton MJ, Lu E, Haddad G, Keplinger AJ, Scepanovic J , Perez-Cervantes C, Chan SSK, Piccirilli JA, Moskowitz IP, Ruthenburg AJ TITLE The cardiac lncRNA Chantico directly regulates Cxcl1 chemokine transcription JOURNAL bioRxiv 2021.12.23.474040 REFERENCE 2 (bases 1 to 3811) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 2021.12.23.474040" FEATURES Location/Qualifiers source 1..3811 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(1151..1169) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1188..1204) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1188..1204) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1201..1223) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1212..1228 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(1236..1266) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1281..1302 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1419..1436) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1590..2178) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1670..1689) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2349..3209) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 2972..2991 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3210..3314) /gene="bla" /label=AmpR promoter primer_bind 3382..3400 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3438..3460) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3560..3579 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3773..3795 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3787..3804 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3788..3804 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 taatacgact cactataggg cgtcattcag taaatataaa tgcagatagc ctacgttctg 61 aaagtggggt tctagggtcc cttggttagt tttaaaaatt caaagtcact ttgagttttg 121 ttttgttttg ttttgtttta aagcttgaga gtggccgtgc agagcaaagt tgtccaatta 181 agtttatatc agtgacagat gggcttatca gaccttcgct ctcagacagt agccatggaa 241 cacttagctg ctgcctcccg aaagtgcaag cagaccagtt caggatctga agctttgttt 301 tggtttgact taaatgtaag tgcaaacagt cgctgtgggg tgtgtgtgtg tgtgtgtgtg 361 tgatactggc cgatgcaggc tgagagtagg ggtgtctcac tcctgactgc gtactccccc 421 tctgttttac acaggcttat ttactgttac acggagactt ttcttcgaac tgttaaactc 481 ttggctgagt ttgtgtggag tatgaggcaa atctctaacc gcaagcatgc agtggaaaga 541 ttaggcatac ttttgaaacg agtttgtgta ccagatcata ttcttggctc aagaaagaaa 601 aagaaagggg gctggggaaa ggcttgtggt tgctactctg ggaactaaag aattcaaaag 661 gcttgttctc catgtttcgt agacacagac atctggaatt gttgttttaa aattacaagc 721 aattgtgtat gctttgttac tgcaacactt attcagaggc aaggacatag gtgccactgt 781 tgtcagaaag ccagatgtcc tttccttaaa ttggcctgcc tggctgctgg cgcctgggtg 841 tcagcagaca ctttgggaat ctcagttagc ggtactgcat ggatatagtg catctgcttc 901 tcacacttct gccaaccttg tgtattggtt tgcttctctc catggtgtta aaacaatgac 961 tgaaagcgtc ttgggtaggg caagggtata tttagcttat agttgcacat gacagtccat 1021 cgtcaaggga gagcatggcg ggaactcaag gactttgagg aggaactgaa gcacaggcat 1081 agaggctgtt cctcactggc ttgctcccca gggcttgctc agctagctgt cttacagagc 1141 ccagaacctc cctttagtga gggttaatta agcttggcgt aatcatggtc atagctgttt 1201 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 1261 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 1321 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 1381 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 1441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 1501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 1561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 1621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 1681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 1741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 1801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 1861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 1921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 1981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 2041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 2101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 2161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 2221 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 2281 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 2341 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 2401 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 2461 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 2521 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 2581 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 2641 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 2701 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 2761 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 2821 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 2881 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 2941 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 3001 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 3061 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 3121 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 3181 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 3241 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 3301 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt 3361 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc 3421 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 3481 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 3541 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 3601 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 3661 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 3721 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 3781 cgacgttgta aaacgacggc cagtgaattc g //

TFORF3026

144502

Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens.

Feng Zhang

Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/407370/0d9a6a72-6932-4715-809e-4c9c42867d91/addgene-plasmid-144502-sequence-407370.gbk

full

LOCUS sequence-407370- 9493 bp DNA circular SYN 25-AUG-2024 DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9493) AUTHORS Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F TITLE A transcription factor atlas of directed differentiation. JOURNAL Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. PUBMED 36608654 REFERENCE 2 (bases 1 to 9493) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5"; volume: "186"; issue: "1"; pages: "209-229.e26" FEATURES Location/Qualifiers source 1..9493 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 5..231 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 232..412 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 459..584 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1077..1310 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1495..1539 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 1770..1980 /label=SV40 promoter /note="SV40 early promoter" promoter 1792..1988 /label=SV40 promoter /note="SV40 early promoter" rep_origin 1839..1974 /label=SV40 ori /note="SV40 origin of replication" CDS 2020..2622 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS" primer_bind complement(2020..2039) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2516..2536 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 2665..2782 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2920..4098 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3151..4089 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4046..4066 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 4118..4142 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(5166..5190) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5198..5239 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 5350..5938 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5403..5423) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5821..5832) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6010..6243 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6321..6442 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6358..6377) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6412..6431 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6482..6617 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(6637..6656) /label=T7 /note="T7 promoter, forward primer" promoter complement(6638..6656) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6666..6683) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6666..6682) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6675..6697) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6824..7279 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6911..6930) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7121..7142 /label=F1ori-F /note="F1 origin, forward primer" promoter 7305..7409 /gene="bla" /label=AmpR promoter CDS 7410..8270 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(7628..7647) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8441..9029 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8930..8949 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9183..9200 /label=L4440 /note="L4440 vector, forward primer" protein_bind 9317..9338 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9353..9383 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9391..9407 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 9396..9418 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 9415..9431 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 9415..9431 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 9450..9470 /label=T3 /note="T3 promoter, forward primer" promoter 9452..9470 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa 61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac 121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa 181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc 241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa 1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca 1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg 2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag 2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc 2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca 2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg 2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc 2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg 2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc 3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc 3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt 3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct 3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac 3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct 3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc 3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt 3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg 3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt 3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga 3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc 3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg 3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg 3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt 3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt 3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg 3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt 4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt 4081 ccatttcagg tgtcgtgagg ctagcatcga ttgatcaaca agtttgtaca aaaaagttgg 4141 catgaccatg gaatctggag ccgagaacca gcagagtgga gatgcagctg taacagaagc 4201 tgaaaaccaa caaatgacag ttcaagccca gccacagatt gccacattag cccaggtatc 4261 tatgccagca gctcatgcaa catcatctgc tcccaccgta actctagtac agctgcccaa 4321 tgggcagaca gttcaagtcc atggagtcat tcaggcggcc cagccatcag ttattcagtc 4381 tccacaagtc caaacagttc agtcttcctg taaggactta aaaagacttt tctccggaac 4441 acagatttca actattgcag aaagtgaaga ttcacaggag tcagtggata gtgtaactga 4501 ttcccaaaag cgaagggaaa ttctttcaag gaggccttcc tacaggaaaa ttttgaatga 4561 cttatcttct gatgcaccag gagtgccaag gattgaagaa gagaagtctg aagaggagac 4621 ttcagcacct gccatcacca ctgtaacggt gccaactcca atttaccaaa ctagcagtgg 4681 acagtatatt gccattaccc agggaggagc aatacagctg gctaacaatg gtaccgatgg 4741 ggtacagggc ctgcaaacat taaccatgac caatgcagca gccactcagc cgggtactac 4801 cattctacag tatgcacaga ccactgatgg acagcagatc ttagtgccca gcaaccaagt 4861 tgttgttcaa gctgcctctg gagacgtaca aacataccag attcgcacag cacccactag 4921 cactattgcc cctggagttg ttatggcatc ctccccagca cttcctacac agcctgctga 4981 agaagcagca cgaaagagag aggtccgtct aatgaagaac agggaagcag ctcgagagtg 5041 tcgtagaaag aagaaagaat atgtgaaatg tttagaaaac agagtggcag tgcttgaaaa 5101 tcaaaacaag acattgattg aggagctaaa agcacttaag gacctttact gccacaaatc 5161 agattaccca actttcttgt acaaagtggt tgatatcggt aagcctatcc ctaaccctct 5221 cctcggtctc gattctacgt agtaatgaac tagtccgtaa cttgaaagta tttcgatttc 5281 ttggctttat atatcttgtg gaaaggacga ggtgtgtatc ctcgacaatt cgggacgcgt 5341 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5401 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5461 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5521 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5581 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5641 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5701 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5761 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5821 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5881 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt 5941 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6001 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta 6061 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 6121 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 6181 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6241 gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat 6301 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca 6361 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 6421 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc 6481 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 6541 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 6601 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg 6661 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt 6721 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc 6781 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc 6841 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 6901 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 6961 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 7021 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 7081 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 7141 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 7201 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 7261 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 7321 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 7381 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 7441 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 7501 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 7561 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 7621 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 7681 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 7741 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 7801 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 7861 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 7921 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 7981 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 8041 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 8101 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 8161 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 8221 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 8281 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 8341 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 8401 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 8461 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 8521 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 8581 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 8641 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 8701 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 8761 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 8821 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 8881 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 8941 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 9001 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 9061 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 9121 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 9181 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg 9241 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca 9301 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact 9361 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa 9421 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc 9481 tggagctgca agc //

pBS532 promoterless GFPcre

11958

Brian Sauer

Le et al Anal Biochem. 1999 Jun 1. 270(2):334-6.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/18819/d0ec9b7f-17c1-4c13-b2e0-acb38041a847/addgene-plasmid-11958-sequence-18819.gbk

partial

LOCUS sequence-18819-e 710 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 710) AUTHORS Le Y, Miller JL, Sauer B TITLE GFPcre fusion vectors with enhanced expression. JOURNAL Anal Biochem. 1999 Jun 1. 270(2):334-6. PUBMED 10334853 REFERENCE 2 (bases 1 to 710) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Anal Biochem. 1999 Jun 1. 270(2):334-6." FEATURES Location/Qualifiers source 1..710 /mol_type="genomic DNA" /organism="unspecified" ORIGIN 1 actatacaaa ggcgccggcg ctggagctgg tgctggcgcc atcatggcac tcgacggaga 61 aagctcaggc tctggctcag agtctgactc catggccaat ttactgaccg tacaccaaaa 121 tttgcctgca ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat 181 gttcagggat cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg 241 gtcgtgggcg gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga 301 tgttcgcgat tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca 361 acatttgggc cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag 421 caatgctgtt tcactggtta tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg 481 tgcaaaacag gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga 541 aaatagcgat cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac 601 cctgttacgt atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg 661 tgggagaatg ttaatccata ttggcagaac gaaaacgctg gttagcaccg //

AT3G47090 L5_pECIA14

114922

Prey vector AT3G47090 L5_pECIA14 should be used with bait vector AT3G47090 L5_pECIA2.

Youssef Belkhadir

Smakowska-Luzan et al Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. Epub 2018 Jan 10.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/288220/e5181fd8-1b48-455f-a0d5-2a92fc9f7a41/addgene-plasmid-114922-sequence-288220.gbk

full

LOCUS sequence-288220- 7079 bp DNA circular SYN 26-AUG-2024 DEFINITION Prey vector AT3G47090 L5_pECIA14 should be used with bait vector AT3G47090 L5_pECIA2.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7079) AUTHORS Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir Y TITLE An extracellular network of Arabidopsis leucine-rich repeat receptor kinases. JOURNAL Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. Epub 2018 Jan 10. PUBMED 29320478 REFERENCE 2 (bases 1 to 7079) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18- 18"; volume: "553"; issue: "7688"; pages: "342-346" FEATURES Location/Qualifiers source 1..7079 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 23..449 /label=MT promoter /note="Drosophila metallothionein promoter" sig_peptide 467..520 /codon_start=1 /product="Drosophila BiP signal sequence" /label=BiP signal sequence /translation="MKLCILLAVVAFVGLSLG" CDS 2375..2398 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4073..4096 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 4112..4129 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4152..4169) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4202..4336 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4226..4245) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4280..4299 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(4480..4496) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4480..4496) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4493..4515) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4504..4520 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4528..4558) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4573..4594 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4711..4728) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4882..5470) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4962..4981) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5641..6501) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6264..6283 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6502..6606) /gene="bla" /label=AmpR promoter primer_bind 6674..6692 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6730..6752) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6852..6871 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind join(7065..7079,1..8) /label=M13/pUC Forward /note="In lacZ gene" ORIGIN 1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact 61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc 121 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat 181 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga 241 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc 301 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc 361 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat 421 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat 481 attactggcc gtcgtggcct ttgttggcct ctcgctcggg accgatgaaa gtgataggca 541 agcgttgttg gagatcaagt ctcaagtttc tgaaagcaaa agggatgcct tgtccgcgtg 601 gaataactca ttccctctct gcagctggaa gtgggttaga tgtggccgca aacacaagag 661 agttactcgt ttggacctcg gaggattgca gttgggagga gtgatatcac catctattgg 721 taatctttcg tttctcatat atcttgacct ctctaataac tcttttggtg gaaccatccc 781 tcaagagatg ggaaacttgt ttagacttaa atatttggct gtgggcttta attatctgga 841 aggagagatt ccagccagtt tgtccaactg ctctagattg ttgtaccttg atttattttc 901 aaacaatctt ggagacggtg ttccttcaga actagggtca ttgagaaagc ttctttatct 961 atatcttggt ctaaacgacc tcaaaggaaa gttccccgta tttatacgaa acttgacgtc 1021 tctaatagtg ctcaacttgg gatataatca tctggaagga gaaattccag atgatatagc 1081 tatgctgagt caaatggtgt ctcttacatt aacaatgaac aacttctctg gcgtttttcc 1141 tcctgctttc tacaacttgt cctcacttga gaatttatac ctccttggta atggtttctc 1201 gggtaacttg aagcctgatt ttggtaatct gctaccaaac attcatgagt tatctttgca 1261 tggaaatttt ctaacaggag ccatcccaac aacacttgcc aatatttcaa ctctggaaat 1321 gtttggaatt ggtaaaaaca ggatgacagg aagtatctct cccaactttg gaaaactaga 1381 gaatttgcat tacctagaac ttgctaacaa ttctttggga agttactctt ttggagatct 1441 tgcatttctt gatgctttga ctaattgctc ccatttgcat gggttatctg tgagttacaa 1501 taggcttggg ggtgctttgc ctacatccat tgtcaatatg tccacggaac tcaccgtttt 1561 gaaccttaaa ggaaacctca tttatggaag cattcctcat gatattggaa atctcatagg 1621 cctacaatca cttttgttgg cagataatct attaacagga ccactcccaa cctctcttgg 1681 aaatcttgta gggctcgggg aattaatttt gttttcaaat aggttttctg gagagatacc 1741 ctcttttata ggcaacctta ctcagttagt aaaactatat ttatccaaca atagttttga 1801 aggaatcgtt cctccgagtc ttggtgattg tagtcacatg ctcgacttac agattgggta 1861 taataagtta aatgggacta tacctaagga gatcatgcaa atcccaaccc ttgttcacct 1921 caacatggaa tctaattctt tgagtggctc tctaccaaac gatattggaa gattacaaaa 1981 tcttgttgag ctattacttg ggaataataa cttatcagga catctcccac aaactttggg 2041 aaaatgtctt tcgatggaag ttatttatct acaagaaaac cattttgatg gaaccattcc 2101 agatataaaa gggttgatgg gtgttaaaaa cgttgatttg tcaaacaata atctctctgg 2161 aagtatatct gaatattttg aaaacttttc caagttggaa tatctcaatc tatccgataa 2221 caatttcgag ggaagagtcc caacagaagg aatatttcaa aatgctactt tagtttcagt 2281 attcggaaac aaaaatcttt gtggaagcat caaggaactg aaactaaagc catgcattgc 2341 ccaagcacca ccagtggaga caaggcatcc atctctggag gtgctgttcc agggaccaag 2401 cggcaatccc ggcggcgatc tggcccccca gatgctgcgc gagctgcagg agaccaatgc 2461 cgccctgcag gacgtgcgcg agctgctgcg ccaccgcgtg aaggagatca ccttcctgaa 2521 gaacaccgtg atggagtgcg acgcctgcgg catgcagccc tctagaggct ccggctccat 2581 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg 2641 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg 2701 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga 2761 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa 2821 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg 2881 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg 2941 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa 3001 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc 3061 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga 3121 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg 3181 tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta 3241 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg 3301 ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc 3361 tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga 3421 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag 3481 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga 3541 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc 3601 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt 3661 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa 3721 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct 3781 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca 3841 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc 3901 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt 3961 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg 4021 caccaccgac gccgcgcacc cgggttctag aggaagcacc tccggcgata acgattacaa 4081 ggatgatgat gataaggcca gcggcaccgg tcatcatcac catcaccatt gagtttaaac 4141 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga 4201 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg 4261 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag 4321 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga 4381 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga 4441 tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 4621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 4741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 4801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 5041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 5101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 5161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 5221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6601 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 6661 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 6721 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 6781 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 6841 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 6901 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 6961 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 7021 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt //

pGBW-m4046468

145496

Yeast Expression plasmid for SARS-CoV-2 nsp9

SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/287673/a9315fa1-5b10-42ab-a535-992e747a19e8/addgene-plasmid-145496-sequence-287673.gbk

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LOCUS sequence-287673- 5193 bp DNA circular SYN 26-AUG-2024 DEFINITION Yeast Expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5193) TITLE SARS-CoV-2 protein Yeast expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5193) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5193 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 25..466 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 25..142 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 437..460 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" CDS 820..837 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 838..903 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" primer_bind complement(948..966) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 1317..2120 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin 2203..3185 /label=2u ori /note="yeast 2u plasmid origin of replication" terminator 3263..3292 /label=T3Te terminator /note="phage T3 early transcription terminator" rep_origin 3314..3901 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" terminator complement(3913..3940) /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(3964..4155) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4156..5016) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4779..4798 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5017..5119) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" terminator complement(5120..5151) /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg 61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg 121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat 181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta 241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag 301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt 361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta 421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg 481 aacaatgagc tttctcctgt tgctttacgg cagatgtcct gcgcagctgg aacaacgcaa 541 actgcctgta ctgacgataa cgctctagct tactataaca ccactaaagg tggtagattc 601 gtcttggcct tgttgtcaga cctgcaagat ttgaagtggg cgagatttcc aaagagcgat 661 ggcactggta ccatatacac cgaattagaa ccaccctgtc gtttcgtgac cgacactcca 721 aaaggtccga aggttaagta tttgtacttt attaagggtt taaataactt gaatcgaggt 781 atggttctgg ggtctttggc tgctacagtc agactccaac tcgtgccgag aggttctgat 841 tataaagatc atgatggtga ttataaagat catgatattg attataaaga tgatgatgat 901 aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat 961 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc 1021 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc 1081 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac 1141 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg 1201 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc 1261 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt 1321 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat 1381 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca 1441 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg 1501 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg 1561 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca 1621 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac 1681 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa 1741 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg 1801 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct 1861 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac 1921 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg 1981 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa 2041 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa 2101 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac 2161 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt 2221 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc 2281 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt 2341 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc 2401 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa 2461 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag 2521 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt 2581 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca 2641 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat 2701 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt 2761 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc 2821 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga 2881 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa 2941 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt 3001 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt 3061 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt 3121 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt 3181 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta 3241 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa 3301 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 3361 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 3421 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 3481 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 3541 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 3601 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 3661 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 3721 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 3781 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg 3841 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 3901 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc 3961 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc 4021 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag 4081 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt 4141 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 4201 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg 4261 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 4321 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt 4381 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 4441 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 4501 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 4561 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc 4621 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 4681 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 4741 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 4801 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt 4861 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 4921 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 4981 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa 5041 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga 5101 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat 5161 ggaggtcagg tatgatttaa atggtcagtt aga //

NAX_CAGGS_CAS9_VPR_EBFP2

167889

PiggyBac compatible plasmid expressing Cas9-VPR

Nghi Dang et al Synthetic Biology, Volume 6, Issue 1, 2021

https://media.addgene.org/snapgene-media/v2.0.0/sequences/354101/5937e24e-3bed-46b3-a0e2-ed0cfe28db06/addgene-plasmid-167889-sequence-354101.gbk

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LOCUS sequence-354101- 13715 bp DNA circular SYN 26-AUG-2024 DEFINITION PiggyBac compatible plasmid expressing Cas9-VPR. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13715) AUTHORS Nghi Dang , Alissa Lance-Byrne, Angela Tung, Xiaoge Guo, Ryan J Cecchi, Joanna Buchthal, Alejandro Chavez, Nan Cher Yeo TITLE Generation and application of a versatile CRISPR toolkit for mammalian cell engineering JOURNAL Synthetic Biology, Volume 6, Issue 1, 2021 REFERENCE 2 (bases 1 to 13715) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Synthetic Biology, Volume 6, Issue 1, 2021" FEATURES Location/Qualifiers source 1..13715 /mol_type="other DNA" /organism="synthetic DNA construct" intron 214..1230 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1154..1176 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1238..1257 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1327..1336 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 1333..5436 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 1333..5436 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 1333..5436 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 5449..5469 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" protein_bind 5506..5530 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5560..5709 /codon_start=1 /product="tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)" /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" CDS 5734..5754 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6184..6540 /codon_start=1 /product="transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)" /label=RelA (p65) AD /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD EDFSSIADMDFSALL" CDS 6559..7128 /codon_start=1 /product="transcriptional activation domain from the human herpesvirus 4 (Epstein-Barr virus) replication and transcription activator Rta/BRLF1 (Hardwick et al., 1992; Chavez et al., 2015)" /label=Rta AD /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL DTFLNDECLLHAMHISTGLSIFDTSLF" primer_bind complement(7153..7172) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7197..7245 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" promoter 7477..7688 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 7698..7707 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 7704..8423 /codon_start=1 /product="enhanced blue variant of GFP (Ai et al., 2007)" /label=EBFP2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL EFRTAAGITLGMDELYK" primer_bind complement(7749..7770) /label=EGFP-N /note="EGFP, reverse primer" polyA_signal 8427..8548 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8464..8483) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8518..8537 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 9311..9345 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind complement(9836..9852) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9836..9852) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(9849..9871) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 9860..9876 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(9884..9914) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9929..9950 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(10067..10084) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10238..10826) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10318..10337) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10997..11857) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 11620..11639 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(11858..11962) /gene="bla" /label=AmpR promoter rep_origin complement(11988..12443) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(12125..12146) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 12337..12356 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 12570..12592 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 12584..12601 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 12585..12601 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" repeat_region 12731..12793 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" enhancer 13214..13593 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 tttttttttt ttttttaatt gtttgggggg gggggggggg gggggggggg gggggggcgc 61 gcgcgggggg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg 121 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg 181 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc 241 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc 301 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa 361 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct 421 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg 481 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttttgcg 541 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc 601 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg 661 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc 721 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg 781 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg 841 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc 901 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt 961 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg 1021 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt 1081 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga 1141 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac 1201 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg 1261 tctcatcatt ttggcaaaga atctagatac cctcgtaaag gtctagagct agcggatcga 1321 acccttgcca ccatggacaa gaagtactcc attgggctcg atatcggcac aaacagcgtc 1381 ggctgggccg tcattacgga cgagtacaag gtgccgagca aaaaattcaa agttctgggc 1441 aataccgatc gccacagcat aaagaagaac ctcattggcg ccctcctgtt cgactccggg 1501 gagacggccg aagccacgcg gctcaaaaga acagcacggc gcagatatac ccgcagaaag 1561 aatcggatct gctacctgca ggagatcttt agtaatgaga tggctaaggt ggatgactct 1621 ttcttccata ggctggagga gtcctttttg gtggaggagg ataaaaagca cgagcgccac 1681 ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat 1741 catctgagga agaagcttgt agacagtact gataaggctg acttgcggtt gatctatctc 1801 gcgctggcgc atatgatcaa atttcgggga cacttcctca tcgaggggga cctgaaccca 1861 gacaacagcg atgtcgacaa actctttatc caactggttc agacttacaa tcagcttttc 1921 gaagagaacc cgatcaacgc atccggagtt gacgccaaag caatcctgag cgctaggctg 1981 tccaaatccc ggcggctcga aaacctcatc gcacagctcc ctggggagaa gaagaacggc 2041 ctgtttggta atcttatcgc cctgtcactc gggctgaccc ccaactttaa atctaacttc 2101 gacctggccg aagatgccaa gcttcaactg agcaaagaca cctacgatga tgatctcgac 2161 aatctgctgg cccagatcgg cgaccagtac gcagaccttt ttttagcggc aaagaacctg 2221 tcagacgcca ttctgctgag tgatattctg cgagtgaaca cggagatcac caaagctccg 2281 ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc aagacttgac tttgctgaag 2341 gcccttgtca gacagcaact gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa 2401 aatggctacg ccggatacat tgacggcgga gcaagccagg aggaatttta caaatttatt 2461 aagcccatct tggaaaaaat ggacggcacc gaggagctgc tggtaaagct taacagagaa 2521 gatctgttgc gcaaacagcg cactttcgac aatggaagca tcccccacca gattcacctg 2581 ggcgaactgc acgctatcct caggcggcaa gaggatttct accccttttt gaaagataac 2641 agggaaaaga ttgagaaaat cctcacattt cggataccct actatgtagg ccccctcgcc 2701 cggggaaatt ccagattcgc gtggatgact cgcaaatcag aagagaccat cactccctgg 2761 aacttcgagg aagtcgtgga taagggggcc tctgcccagt ccttcatcga aaggatgact 2821 aactttgata aaaatctgcc taacgaaaag gtgcttccta aacactctct gctgtacgag 2881 tacttcacag tttataacga gctcaccaag gtcaaatacg tcacagaagg gatgagaaag 2941 ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg acctcctctt caagacgaac 3001 cggaaagtta ccgtgaaaca gctcaaagaa gactatttca aaaagattga atgtttcgac 3061 tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat 3121 ctcctgaaaa tcattaaaga caaggacttc ctggacaatg aggagaacga ggacattctt 3181 gaggacattg tcctcaccct tacgttgttt gaagataggg agatgattga agaacgcttg 3241 aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac agctcaagag gcgccgatat 3301 acaggatggg ggcggctgtc aagaaaactg atcaatggga tccgagacaa gcagagtgga 3361 aagacaatcc tggattttct taagtccgat ggatttgcca accggaactt catgcagttg 3421 atccatgatg actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag 3481 ggggacagtc ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga 3541 atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc 3601 gagaatatcg ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac 3661 agtagggaaa ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt 3721 aaggaacacc cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg 3781 cagaacggca gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac 3841 gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg 3901 ttgacaagat ccgataaaaa tagagggaag agtgataacg tcccctcaga agaagttgtc 3961 aagaaaatga aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag 4021 ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc 4081 atcaaaaggc agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat 4141 tcacgcatga acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt 4201 actctgaagt ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga 4261 gagatcaaca attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca 4321 cttatcaaaa aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac 4381 gatgttagga aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac 4441 ttcttttaca gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag 4501 attcggaagc gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag 4561 ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa 4621 aagaccgaag tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc 4681 gacaagctga tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct 4741 cctacagtcg cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa 4801 ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa 4861 aaccccatcg actttctcga ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt 4921 aagcttccca agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt 4981 gcgggcgagc tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg 5041 tatctggcca gccactatga aaagctcaaa gggtctcccg aagataatga gcagaagcag 5101 ctgttcgtgg aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgaattc 5161 tccaaaagag tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag 5221 cacagggata agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc 5281 aacttgggcg cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac 5341 acctctacaa aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat 5401 gaaacaagaa tcgacctctc tcagctcggt ggagacagca gggctgaccc caagaagaag 5461 aggaaggtgt cgccagggat ccgtcgactt gacgcgttaa tatcaacaag tttgtacaaa 5521 aaagcaggct acaaagaggc cagcggttcc ggacgggctg acgcattgga cgattttgat 5581 ctggatatgc tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat 5641 gcccttgatg actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg 5701 gacatgctga ttaactctag aagttccgga tctccgaaaa agaaacgcaa agttggtagc 5761 cagtacctgc ccgacaccga cgaccggcac cggatcgagg aaaagcggaa gcggacctac 5821 gagacattca agagcatcat gaagaagtcc cccttcagcg gccccaccga ccctagacct 5881 ccacctagaa gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa acctgccccc 5941 cagccttacc ccttcaccag cagcctgagc accatcaact acgacgagtt ccctaccatg 6001 gtgttcccca gcggccagat ctctcaggcc tctgctctgg ctccagcccc tcctcaggtg 6061 ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg tgtctgcact ggctcaggca 6121 ccagcacccg tgcctgtgct ggctcctgga cctccacagg ctgtggctcc accagcccct 6181 aaacctacac aggccggcga gggcacactg tctgaagctc tgctgcagct gcagttcgac 6241 gacgaggatc tgggagccct gctgggaaac agcaccgatc ctgccgtgtt caccgacctg 6301 gccagcgtgg acaacagcga gttccagcag ctgctgaacc agggcatccc tgtggcccct 6361 cacaccaccg agcccatgct gatggaatac cccgaggcca tcacccggct cgtgacaggc 6421 gctcagaggc ctcctgatcc agctcctgcc cctctgggag caccaggcct gcctaatgga 6481 ctgctgtctg gcgacgagga cttcagctct atcgccgata tggatttctc agccttgctg 6541 ggctctggca gcggcagccg ggattccagg gaagggatgt ttttgccgaa gcctgaggcc 6601 ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa acgaatccgg 6661 ccatttcatc ctccaggaag tccatgggcc aaccgcccac tccccgccag cctcgcacca 6721 acaccaaccg gtccagtaca tgagccagtc gggtcactga ccccggcacc agtccctcag 6781 ccactggatc cagcgcccgc agtgactccc gaggccagtc acctgttgga ggatcccgat 6841 gaagagacga gccaggctgt caaagccctt cgggagatgg ccgatactgt gattccccag 6901 aaggaagagg ctgcaatctg tggccaaatg gacctttccc atccgccccc aaggggccat 6961 ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct ggactcaccc 7021 ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg cctcttgcat 7081 gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgttttg agtttaaacg 7141 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc 7201 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc 7261 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc 7321 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag 7381 ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg aaatcggatc cgcggccgca 7441 ctgattctag ctgcagaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag 7501 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg 7561 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag 7621 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca 7681 gaacacagga tcctcgagcc accatggtga gcaagggcga ggagctgttc accggggtgg 7741 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgaggggcg 7801 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca 7861 agctgcccgt gccctggccc accctcgtga ccaccctgag ccacggcgtg cagtgcttcg 7921 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 7981 acgtccagga gcgcaccatc ttcttcaagg acgacggcac ctacaagacc cgcgccgagg 8041 tgaagttcga gggcgacacc ctagtgaacc gcatcgagct gaagggcgtc gacttcaagg 8101 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacatctata 8161 tcatggccgt caagcagaag aacggcatca aggtgaactt caagatccgc cacaacgtgg 8221 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 8281 ccgtgctgct gcccgacagc cactacctga gcacccagtc cgtgctgagc aaagacccca 8341 acgagaagcg cgatcacatg gtcctgctgg agttccgcac cgccgccggg atcactctcg 8401 gcatggacga gctgtacaag taagttaact tgtttattgc agcttataat ggttacaaat 8461 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 8521 gtttgtccaa actcatcaat gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat 8581 tagtctatca gaagctcatc tggtctccct tccgggggac aagacatccc tgtttaatat 8641 ttaaacagca gtgttcccaa actgggttct tatatccctt gctctggtca accaggttgc 8701 agggtttcct gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc caggtttagc 8761 cccggaattg actggattcc ttttttaggg cccattggta tggctttttc cccgtatccc 8821 cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc 8881 caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg 8941 ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa 9001 ttacatccct gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat 9061 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta 9121 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag 9181 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg 9241 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat 9301 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat 9361 caggatcata tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg 9421 ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag 9481 tttgccgagg atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg 9541 attcgggaag gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg 9601 gataccagtt cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc 9661 agcaacccga acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac 9721 agcggtgcgg cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag 9781 aaatggacat ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat 9841 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 9901 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 9961 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 10021 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 10081 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 10141 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 10201 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 10261 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 10321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 10381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 10441 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 10501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 10561 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 10621 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 10681 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 10741 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 10801 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 10861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 10921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 10981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 11041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 11101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 11161 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 11221 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 11281 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 11341 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 11401 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 11461 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 11521 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 11581 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 11641 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 11701 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 11761 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 11821 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 11881 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 11941 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt 12001 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata 12061 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt 12121 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg 12181 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt 12241 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc 12301 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg 12361 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct 12421 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg 12481 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 12541 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 12601 tgagcgcgcc tcgttcattc acgtttttga acccgtggag gacgggcaga ctcgcggtgc 12661 aaatgtgttt tacagcgtga tggagcagat gaagatgctc gacacgctgc agaacacgca 12721 gctagattaa ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgtgta 12781 aaattgacgc atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga 12841 tattaagttt tattatattt acacttacat actaataata aattcaacaa acaatttatt 12901 tatgtttatt tatttattaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa 12961 acttttatga gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc 13021 gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc 13081 cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt 13141 cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa 13201 ggcctccacg gccgacattg attattgact agttattaat agtaatcaat tacggggtca 13261 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 13321 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 13381 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 13441 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 13501 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 13561 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc 13621 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta 13681 ttttgtgcag cgatgggggc gggggggggg ggggg //

TFORF1070

142243

Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens.

Feng Zhang

Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/395740/43285bf4-8706-4c82-b116-a52806018c94/addgene-plasmid-142243-sequence-395740.gbk

full

LOCUS sequence-395740- 11572 bp DNA circular SYN 26-AUG-2024 DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11572) AUTHORS Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F TITLE A transcription factor atlas of directed differentiation. JOURNAL Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. PUBMED 36608654 REFERENCE 2 (bases 1 to 11572) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5"; volume: "186"; issue: "1"; pages: "209-229.e26" FEATURES Location/Qualifiers source 1..11572 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 5..231 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 232..412 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 459..584 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1077..1310 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1495..1539 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 1770..1980 /label=SV40 promoter /note="SV40 early promoter" promoter 1792..1988 /label=SV40 promoter /note="SV40 early promoter" rep_origin 1839..1974 /label=SV40 ori /note="SV40 origin of replication" CDS 2020..2622 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS" primer_bind complement(2020..2039) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2516..2536 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 2665..2782 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2920..4098 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3151..4089 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4046..4066 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" misc_feature 7429..8017 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7482..7502) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7900..7911) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 8089..8322 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8400..8521 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8437..8456) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8491..8510 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 8561..8696 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(8716..8735) /label=T7 /note="T7 promoter, forward primer" promoter complement(8717..8735) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8745..8762) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8745..8761) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8754..8776) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8903..9358 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8990..9009) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9200..9221 /label=F1ori-F /note="F1 origin, forward primer" promoter 9384..9488 /gene="bla" /label=AmpR promoter CDS 9489..10349 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(9707..9726) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10520..11108 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11009..11028 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11262..11279 /label=L4440 /note="L4440 vector, forward primer" protein_bind 11396..11417 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11432..11462 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11470..11486 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 11475..11497 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 11494..11510 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 11494..11510 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 11529..11549 /label=T3 /note="T3 promoter, forward primer" promoter 11531..11549 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa 61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac 121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa 181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc 241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa 1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca 1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg 2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag 2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc 2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca 2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg 2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc 2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg 2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc 3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc 3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt 3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct 3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac 3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct 3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc 3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt 3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg 3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt 3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga 3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc 3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg 3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg 3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt 3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt 3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg 3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt 4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt 4081 ccatttcagg tgtcgtgagg ctagcaccgc caccatgact actgcaaact gtggcgccca 4141 cgacgagctc gacttcaaac tcgtctttgg cgaggacggg gcgccggcgc cgccgccccc 4201 gggctcgcgg cctgcagatc ttgagccaga tgattgtgca tccatttaca tctttaatgt 4261 agatccacct ccatctactt taaccacacc actttgctta ccacatcatg gattaccgtc 4321 tcactcttct gttttgtcac catcgtttca gctccaaagt cacaaaaact atgaaggaac 4381 ttgtgagatt cctgaatcta aatatagccc attaggtggt cccaaaccct ttgagtgccc 4441 aagtattcaa attacatcta tctctcctaa ctgtcatcaa gaattagatg cacatgaaga 4501 tgacctacag ataaatgacc cagaacggga atttttggaa aggccttcta gagatcatct 4561 ctatcttcct cttgagccat cctaccggga gtcttctctt agtcctagtc ctgccagcag 4621 catctcttct aggagttggt tctctgatgc atcttcttgt gaatcgcttt cacatattta 4681 tgatgatgtg gactcagagt tgaatgaagc tgcagcccga tttacccttg gatcccctct 4741 gacttctcct ggtggctctc cagggggctg ccctggagaa gaaacttggc atcaacagta 4801 tggacttgga cactcattat cacccaggca atctccttgc cactctccta gatccagtgt 4861 cactgatgag aattggctga gccccaggcc agcctcagga ccctcatcaa ggcccacatc 4921 cccctgtggg aaacggaggc actccagtgc tgaagtttgt tatgctgggt ccctttcacc 4981 ccatcactca cctgttcctt cacctggtca ctcccccagg ggaagtgtga cagaagatac 5041 gtggctcaat gcttctgtcc atggtgggtc aggccttggc cctgcagttt ttccatttca 5101 gtactgtgta gagactgaca tccctctcaa aacaaggaaa acttctgaag atcaagctgc 5161 catactacca ggaaaattag agctgtgttc agatgaccaa gggagtttat caccagcccg 5221 ggagacttca atagatgatg gccttggatc tcagtatcct ttaaagaaag attcatgtgg 5281 tgatcagttt ctttcagttc cttcaccctt tacctggagc aaaccaaagc ctggccacac 5341 ccctatattt cgcacatctt cattacctcc actagactgg cctttaccag ctcattttgg 5401 acaatgtgaa ctgaaaatag aagtgcaacc taaaactcat catcgagccc attatgaaac 5461 tgaaggtagc cgaggggcag taaaagcatc tactggggga catcctgttg tgaagctcct 5521 gggctataac gaaaagccaa taaatctaca aatgtttatt gggacagcag atgatcgata 5581 tttacgacct catgcatttt accaggtgca tcgaatcact gggaagacag tcgctactgc 5641 aagccaagag ataataattg ccagtacaaa agttctggaa attccacttc ttcctgaaaa 5701 taatatgtca gccagtattg attgtgcagg tattttgaaa ctccgcaatt cagatataga 5761 acttcgaaaa ggagaaactg atattggcag aaagaatact agagtacgac ttgtgtttcg 5821 tgtacacatc ccacagccca gtggaaaagt cctttctctg cagatagcct ctatacccgt 5881 tgagtgctcc cagcggtctg ctcaagaact tcctcatatt gagaagtaca gtatcaacag 5941 ttgttctgta aatggaggtc atgaaatggt tgtgactgga tctaattttc ttccagaatc 6001 caaaatcatt tttcttgaaa aaggacaaga tggacgacct cagtgggagg tagaagggaa 6061 gataatcagg gaaaaatgtc aaggggctca cattgtcctt gaagttcctc catatcataa 6121 cccagcagtt acagctgcag tgcaggtgca cttttatctt tgcaatggca agaggaaaaa 6181 aagccagtct caacgtttta cttatacacc agttttgatg aagcaagaac acagagaaga 6241 gattgatttg tcttcagttc catctttgcc tgtgcctcat cctgctcaga cccagaggcc 6301 ttcctctgat tcagggtgtt cacatgacag tgtactgtca ggacagagaa gtttgatttg 6361 ctccatccca caaacatatg catccatggt gacctcatcc catctgccac agttgcagtg 6421 tagagatgag agtgttagta aagaacagca tatgattcct tctccaattg tacaccagcc 6481 ttttcaagtc acaccaacac ctcctgtggg gtcttcctat cagcctatgc aaactaatgt 6541 tgtgtacaat ggaccaactt gtcttcctat taatgctgcc tctagtcaag aatttgattc 6601 agttttgttt cagcaggatg caactctttc tggtttagtg aatcttggct gtcaaccact 6661 gtcatccata ccatttcatt cttcaaattc aggctcaaca ggacatctct tagcccatac 6721 acctcattct gtgcataccc tgcctcatct gcaatcaatg ggatatcatt gttcaaatac 6781 aggacaaaga tctctttctt ctccagtggc tgaccagatt acaggtcagc cttcgtctca 6841 gttacaacct attacatatg gtccttcaca ttcagggtct gctacaacag cttccccagc 6901 agcttctcat cccttggcta gttcaccgct ttctgggcca ccatctcctc agcttcagcc 6961 tatgccttac caatctccta gctcaggaac tgcctcatca ccgtctccag ccaccagaat 7021 gcattctgga cagcactcaa ctcaagcaca aagtacgggc caggggggtc tttctgcacc 7081 ttcatcctta atatgtcaca gtttgtgtga tccagcgtca tttccacctg atggggcaac 7141 tgtgagcatt aaacctgaac cagaagatcg agagcctaac tttgcaacca ttggtctgca 7201 ggacatcact ttagatgatg accaatttat atctgacttg gaacaccagc catcaggttc 7261 agcagagaaa tggcctaacc acagtgtgct ctcatgtcca gctcctttct ggagaatcta 7321 gtaatgaact agtccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg 7381 aaaggacgac tagtggaact ccttcgccgc gtcacgcgtt aagtcgacaa tcaacctctg 7441 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 7501 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 7561 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 7621 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 7681 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 7741 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 7801 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 7861 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 7921 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 7981 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt 8041 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa 8101 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc 8161 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 8221 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 8281 gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat 8341 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga 8401 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 8461 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 8521 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc 8581 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 8641 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 8701 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta 8761 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc 8821 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg 8881 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 8941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 9001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 9061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 9121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 9181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 9241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 9301 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag 9361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 9421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 9481 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 9541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 9601 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 9661 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 9721 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 9781 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 9841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 9901 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 9961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 10021 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 10081 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 10141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 10201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 10261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 10321 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 10381 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 10441 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 10501 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 10561 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 10621 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 10681 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 10741 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 10801 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 10861 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 10921 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 10981 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 11041 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 11101 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 11161 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 11221 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 11281 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 11341 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg 11401 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt 11461 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg 11521 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gc //

FUGW-Mito-TurboID

200963

3rd gen lentiviral plasmid for expressing mitochondria localized TurboID with EGFP

Keriann Backus

Yan et al Cell Chem Biol. 2023 Jul 20;30(7):811-827.e7. doi: 10.1016/j.chembiol.2023.06.008. Epub 2023 Jul 6.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/393511/210329f6-663a-40ae-8a74-c55b2d1a9e85/addgene-plasmid-200963-sequence-393511.gbk

full

LOCUS sequence-393511- 11117 bp DNA circular SYN 25-AUG-2024 DEFINITION 3rd gen lentiviral plasmid for expressing mitochondria localized TurboID with EGFP. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11117) AUTHORS Yan T, Julio AR, Villanueva M, Jones AE, Ball AB, Boatner LM, Turmon AC, Nguyen KB, Yen SL, Desai HS, Divakaruni AS, Backus KM TITLE Proximity-labeling chemoproteomics defines the subcellular cysteinome and inflammation-responsive mitochondrial redoxome. JOURNAL Cell Chem Biol. 2023 Jul 20;30(7):811-827.e7. doi: 10.1016/j.chembiol.2023.06.008. Epub 2023 Jul 6. PUBMED 37419112 REFERENCE 2 (bases 1 to 11117) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.chembiol.2023.06.008"; journalName: "Cell Chem Biol"; date: "2023-07-20- 20"; volume: "30"; issue: "7"; pages: "811-827.e7" FEATURES Location/Qualifiers source 1..11117 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 117..496 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 498..696 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 651..671 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 714..894 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 941..1066 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1559..1792 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1977..2021 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2319..2436 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2501..3712 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3549..3568 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3908..4864 /codon_start=1 /product="mutant of the E. coli biotin ligase BioID engineered for improved catalytic efficiency (Branon et al., 2018)" /label=TurboID /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK" CDS 4871..4912 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" regulatory 4922..4931 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 4928..5647 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4928..5644 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5234..5253) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5581..5602 /label=EGFP-C /note="EGFP, forward primer" misc_feature 5688..6276 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5741..5761) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6159..6170) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(6279..6295) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6280..6296) /label=pBluescriptKS /note="For pBluescript vector" LTR 6801..6981 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(7007..7024) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 7013..7237 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 7283..7711 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7370..7389) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7580..7601 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7720..7740) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7725..8054 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7905..8040 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7967..7986 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 8102..8149 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 8168..8542 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 8672..8793 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8709..8728) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8763..8782 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(8842..8858) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8842..8858) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8855..8877) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8866..8882 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(8890..8920) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8935..8956 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(9073..9090) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9244..9832) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9324..9343) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10003..10863) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(10864..10968) /gene="bla" /label=AmpR promoter primer_bind complement(11043..11062) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 61 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 121 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 181 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 241 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 481 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 541 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 601 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 661 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc 721 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 781 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 841 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 901 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg 961 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat 1021 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg 1081 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa 1141 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct 1201 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac 1261 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca 1321 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa 1381 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga 1441 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag 1501 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag 1561 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga 1621 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc 1681 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc 1741 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt 1801 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta 1861 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta 1921 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga 1981 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa 2041 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa 2101 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat 2161 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag 2221 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg 2281 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt 2341 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa 2401 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga 2461 gatccagttt ggttaattaa cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg 2521 gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc 2581 agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac 2641 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct 2701 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc 2761 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg 2821 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat 2881 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg 2941 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc 3001 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc 3061 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg 3121 gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct 3181 tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga 3241 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca 3301 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt 3361 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg 3421 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg 3481 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct 3541 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga 3601 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta 3661 gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acgaagcttg 3721 ggctgcaggt cgactctaga ggatccgggc cgccaccatg ctggccaccc gcgtgttcag 3781 cctggtgggc aagcgcgcca tcagcaccag cgtgtgcgtg cgcgcccacg gatcaggaag 3841 cggcggccgc ggatcaggaa gcggatcagg aagcggatca ggaagcggat caggaagcgg 3901 atcaggaaaa gacaatactg tgcctctgaa gctgatcgct ctcctggcta atggcgagtt 3961 ccatagtggc gaacagctgg gagaaaccct gggcatgtcc agggccgcta tcaacaagca 4021 cattcagact ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa agggctactc 4081 tctgcccgag cctatcccgc tgctgaacgc taaacagatt ctgggacagc tggacggcgg 4141 gagcgtggca gtcctgcctg tggtcgactc caccaatcag tacctgctgg atcgaatcgg 4201 cgagctgaag agtggggatg cttgcattgc agaatatcag caggcaggga gaggaagcag 4261 agggaggaaa tggttctctc cttttggagc taacctgtac ctgagtatgt tttggcgcct 4321 gaagcgggga ccagcagcaa tcggcctggg cccggtcatc ggaattgtca tggcagaagc 4381 gctgcgaaag ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca 4441 ggatagaaag ctggcaggca tcctggtgga gctggccgga ataacaggcg atgctgcaca 4501 gatcgtcatt ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa 4561 tcagggctgg atcacactgc aggaagcagg gattaacctg gacaggaata ctctggccgc 4621 tacgctgatc cgagagctgc gggcagccct ggaactgttc gagcaggaag gcctggctcc 4681 atatctgcca cggtgggaga agctggataa cttcatcaat agacccgtga agctgatcat 4741 tggggacaaa gagattttcg ggattagccg ggggattgat aaacagggag ccctgctgct 4801 ggaacaggac ggagttatca aaccctggat gggcggagaa atcagtctgc ggtctgccga 4861 aaaggctagc ggcaagccca tccccaaccc cctgctgggc ctggacagca ccggaccggt 4921 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga 4981 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc 5041 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg 5101 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca 5161 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac 5221 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga 5281 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct 5341 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca 5401 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca 5461 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 5521 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca 5581 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 5641 caagtaaagc ggccgcgact ctagaattcg atatcaagct tatcgataat caacctctgg 5701 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 5761 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 5821 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 5881 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 5941 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 6001 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 6061 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca 6121 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 6181 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 6241 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc 6301 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 6361 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 6421 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 6481 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 6541 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac 6601 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca 6661 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 6721 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag 6781 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct 6841 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 6901 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6961 cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct 7021 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 7081 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 7141 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 7201 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 7261 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 7321 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 7381 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 7441 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 7501 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 7561 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 7621 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 7681 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 7741 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 7801 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 7861 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 7921 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 7981 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 8041 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg 8101 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa 8161 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg 8221 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt 8281 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt 8341 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg 8401 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg 8461 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt 8521 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta 8581 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 8641 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 8701 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 8761 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 8821 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 8881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 8941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 9001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 9061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 9121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 9181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 9241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 9301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 9361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 9421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 9481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 9541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 9601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 9661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 9721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 9781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 9841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 9901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 9961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 10021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 10081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 10141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 10201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 10261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 10321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 10381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 10441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 10501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 10561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 10621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 10681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 10741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 10801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 10861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 10921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 10981 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc 11041 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg 11101 gaggtcgctg agtagtg //

3C8X

51336

Cheryl Arrowsmith

SGC Plasmids - 2013 (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/79940/72cdec01-ec41-4a6a-ba33-b56ea75f0d6e/addgene-plasmid-51336-sequence-79940.gbk

partial

LOCUS sequence-79940-e 842 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 842) TITLE SGC Plasmids - 2013 JOURNAL Unpublished REFERENCE 2 (bases 1 to 842) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" FEATURES Location/Qualifiers source 1..842 /mol_type="genomic DNA" /organism="unspecified" sig_peptide 81..143 /codon_start=1 /product="signal sequence from honeybee melittin" /label=melittin signal sequence /translation="MKFLVNVALVFMVVYISYIYA" CDS 156..173 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 195..215 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" ORIGIN 1 gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc 61 gcggatctcg gtccgaaacc atgaaattct tagtcaacgt tgcccttgtt tttatggtcg 121 tatacatttc ttacatctat gcggccgctc cggaacatca ccatcaccat cacgattacg 181 atatcccaac gaccgaaaac ctgtattttc agggcgccat ggatgcggcg cagggcaagg 241 aagtggtact gctggacttt gctgcagctg gaggggagct cggctggctc acacacccgt 301 atggcaaagg gtgggacctg atgcagaaca tcatgaatga catgccgatc tacatgtact 361 ccgtgtgcaa cgtgatgtct ggcgaccagg acaactggct ccgcaccaac tgggtgtacc 421 gaggagaggc tgagcgtatc ttcattgagc tcaagtttac tgtacgtgac tgcaacagct 481 tccctggtgg cgccagctcc tgcaaggaga ctttcaacct ctactatgcc gagtcggacc 541 tggactacgg caccaacttc cagaagcgcc tgttcaccaa gattgacacc attgcgcccg 601 atgagatcac cgtcagcagc gacttcgagg cacgccacgt gaagctgaac gtggaggagc 661 gctccgtggg gccgctcacc cgcaaaggct tctacctggc cttccaggat atcggtgcct 721 gtgtggcgct gctctccgtc cgtgtctact acaagaagtg cccctgatga agcttgtcga 781 gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 841 aa //

pFH39

126013

Level0 Rice OsU3 Pol III promoter with transcription Start-A

Vladimir Nekrasov

Hahn et al BMC Plant Biol. 2020 Apr 23;20(1):179. doi: 10.1186/s12870-020-02388-2.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/252798/9913c61d-57be-41ff-8773-5e25a408f9d7/addgene-plasmid-126013-sequence-252798.gbk

full

LOCUS sequence-252798- 2450 bp DNA circular SYN 26-AUG-2024 DEFINITION Level0 Rice OsU3 Pol III promoter with transcription Start-A. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2450) AUTHORS Hahn F, Korolev A, Sanjurjo Loures L, Nekrasov V TITLE A modular cloning toolkit for genome editing in plants. JOURNAL BMC Plant Biol. 2020 Apr 23;20(1):179. doi: 10.1186/s12870-020-02388-2. PUBMED 32326901 REFERENCE 2 (bases 1 to 2450) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC Plant Biol."; date: "2020-04-23"; volume: "20(1):179. doi"; pages: " 10.1186/s12870-020-02388-2" FEATURES Location/Qualifiers source 1..2450 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 285..1076 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 1417..2005 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1906..1925 /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2058..2438 /label=OsU3 promoter /note="Oryza sativa (rice) snRNA U3 promoter" ORIGIN 1 gtgccgaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt 61 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg 121 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg 181 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg 241 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgggg gaagcggtga 301 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac 361 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca 421 gcgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt 481 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg 541 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg 601 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca 661 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg 721 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc 781 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg 841 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg 901 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg 961 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt 1021 tggaagaatt tgtccattac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc 1081 tagctagaaa ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga 1141 tgcactaagc acataattgc tcacagccaa actatcaggt caagtctgct tttattattt 1201 ttaagcgtgc ataataagcc ctacacaaat tgggagatat atcatgctgt cagaccaagt 1261 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 1321 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 1381 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 1441 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 1501 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 1561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 1621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 1681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 1741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 1801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 1861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 1921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 1981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctgca 2041 ctctgtggtc tcaggagaag gaatctttaa acatacgaac agatcactta aagttcttct 2101 gaagcaactt aaagttatca ggcatgcatg gatcttggag gaatcagatg tgcagtcagg 2161 gaccatagca caagacaggc gtcttgtact ggtgctacca gcaaatgctg gaagccggga 2221 acactgggta cgttggaaac cacgtgatgt gaagaagtaa gataaactgt aggagaaaag 2281 catttcgtag tgggccatga agcctttcag gacatgtatt gcagtatggg ccggcccatt 2341 acgcaattgg acgacaacaa agactagtat tagtaccacc tcggctatcc acatagatca 2401 aagctgattt aaaagagttg tgcagatgat ccgtggcacg agaccacgaa //

AT1G66830 X005_pECIA14

114810

Prey vector AT1G66830 X005_pECIA14 should be used with bait vector AT1G66830 X005_pECIA2.

Youssef Belkhadir

Smakowska-Luzan et al Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. Epub 2018 Jan 10.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/237787/10605172-b88e-48df-b8fa-0f9a32a9b6c3/addgene-plasmid-114810-sequence-237787.gbk

full

LOCUS sequence-237787- 6071 bp DNA circular SYN 26-AUG-2024 DEFINITION Prey vector AT1G66830 X005_pECIA14 should be used with bait vector AT1G66830 X005_pECIA2.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6071) AUTHORS Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir Y TITLE An extracellular network of Arabidopsis leucine-rich repeat receptor kinases. JOURNAL Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. Epub 2018 Jan 10. PUBMED 29320478 REFERENCE 2 (bases 1 to 6071) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18- 18"; volume: "553"; issue: "7688"; pages: "342-346" FEATURES Location/Qualifiers source 1..6071 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 40..58 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(96..118) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 218..237 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 431..453 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 446..462 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 468..894 /label=MT promoter /note="Drosophila metallothionein promoter" sig_peptide 912..965 /codon_start=1 /product="Drosophila BiP signal sequence" /label=BiP signal sequence /translation="MKLCILLAVVAFVGLSLG" CDS 1812..1835 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 3510..3533 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 3549..3566 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3589..3606) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3639..3773 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3663..3682) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3717..3736 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3917..3933) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3917..3933) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3930..3952) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3941..3957 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3965..3995) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4010..4031 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4148..4165) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4319..4907) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4399..4418) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5078..5938) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5701..5720 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5939..6043) /gene="bla" /label=AmpR promoter ORIGIN 1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg 481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc 541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg 601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat 661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca 721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa 781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg 841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat 901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc 961 tcgggtctct aaacgatcaa ggtcttgctc ttctttcctt taaacaatcc atacaaaacc 1021 agagtgactc tgtttttacc aattggaact cttcagattc aaacccttgt tcttggcaag 1081 gtgttacatg taattatgat atgagagttg tctcgattcg tcttcctaat aaacgtcttt 1141 cgggttctct tgatccatcc attggaagtc tcttgtccct tcgtcacatt aacctaagag 1201 acaatgactt tcaagggaag ttacctgtgg aactctttgg acttaaaggg cttcaaagct 1261 tggtgctttc tggaaattct ttttccgggt ttgtcccaga agagattggt agcttgaaga 1321 gtctaatgac tctggatctt tcggagaatt ctttcaacgg ttcgatttct ttgtcgctta 1381 ttccatgcaa gaagctgaaa acgcttgttc ttagcaagaa cagtttctct ggagatcttc 1441 caacagggtt aggttctaac ttggttcatc ttcggacact gaatctttct ttcaatcgct 1501 tgactggaac gatccccgaa gatgtaggaa gtctggagaa tctgaaaggt acacttgatc 1561 tttctcataa cttcttctca ggtatgattc caacaagtct tggaaatctc cctgagcttc 1621 tctatgtcga tctttcttac aataatctca gtggtccgat accaaaattc aacgttctgt 1681 tgaatgccgg tccaaatgct ttccaaggga accctttcct ctgtggatta ccgataaaga 1741 tttcttgcag cacaagaaac acacaagtag ttccttctca actatataca agaagagcca 1801 accatcattc tctggaggtg ctgttccagg gaccaagcgg caatcccggc ggcgatctgg 1861 ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc cctgcaggac gtgcgcgagc 1921 tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa caccgtgatg gagtgcgacg 1981 cctgcggcat gcagccctct agaggctccg gctccatcat cccagttgag gaggagaacc 2041 cggacttctg gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg 2101 cacagacagc cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg 2161 tgacagctgc caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc 2221 tggccatgga ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg 2281 tgccagacag tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga 2341 ccattggctt gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg 2401 tcatctccgt gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca 2461 cacgagtgca gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt 2521 actcggacgc cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc 2581 agctcatctc caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc 2641 gcatgggaac cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg 2701 acgggaagaa tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga 2761 accgcactga gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct 2821 ttgagcctgg agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga 2881 tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct 2941 tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga 3001 ctgagacgat catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg 3061 acacgctgag cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc 3121 tgcgagggag ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca 3181 cggtcctcct atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg 3241 ttaccgagag cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg 3301 aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg 3361 ttcacggcgt gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg 3421 agccctacac cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg 3481 gttctagagg aagcacctcc ggcgataacg attacaagga tgatgatgat aaggccagcg 3541 gcaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc 3601 cttctaaggc ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca 3661 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa 3721 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca 3781 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca 3841 agtaaaacct ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa 3901 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 3961 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 4021 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4081 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 4141 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4201 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 4261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 4561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 4621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 4681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 4741 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 4801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 4861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4981 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 5041 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 5101 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 5161 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 5221 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5281 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5341 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5401 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5461 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5521 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5581 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5641 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5701 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5761 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5821 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5881 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5941 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 6001 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 6061 tgccacctga c //

pCMV2-SMAC-HA-eGFP

67488

Targets HA-eGFP to mitochondrial intermembrane space (IMS)

Richard Kahn

Kerr et al Cell Logist. 2015 Sep 21;5(3):e1090523. doi: 10.1080/21592799.2015.1090523. eCollection 2015 Jul-Sep.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/118321/75678ac9-55b3-4823-9b38-01cc261c253b/addgene-plasmid-67488-sequence-118321.gbk

partial

LOCUS sequence-118321- 880 bp DNA linear UNA 26-AUG-2024 DEFINITION Targets HA-eGFP to mitochondrial intermembrane space (IMS). ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 880) AUTHORS Kerr SC, Kahn RA TITLE Tool box: Plasmids for the expression or knockdown of human ARF Family GTPases (ARF/ARL/SAR) and their co-expression in bacteria with N-myristoyltransferases. JOURNAL Cell Logist. 2015 Sep 21;5(3):e1090523. doi: 10.1080/21592799.2015.1090523. eCollection 2015 Jul-Sep. PUBMED 27057421 REFERENCE 2 (bases 1 to 880) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Logist."; date: "2015-09-21"; volume: "5(3):e1090523. doi"; pages: " 10.1080/21592799.2015.1090523. eCollection 2015 Jul-Sep" FEATURES Location/Qualifiers source 1..880 /mol_type="genomic DNA" /organism="unspecified" primer_bind 23..42 /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" CDS complement(67..783) /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS complement(70..783) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(112..133) /label=EGFP-C /note="EGFP, forward primer" primer_bind 461..480 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 720..741 /label=EGFP-N /note="EGFP, reverse primer" CDS complement(790..816) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" ORIGIN 1 ggcttcatga tgtccccata atttttggca gagggaaaaa gatctgctag caagcttctc 61 gagtcattac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc 121 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct 181 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta 241 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt 301 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc 361 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg 421 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc 481 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc 541 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt 601 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag 661 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac 721 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct 781 cacgaattca gcgtaatctg gaacatcgta tgggtaggat ccaataggaa ccgcacacag 841 ggttactcca aagccaatcg tcacagtttt gtgccatggt //

pDONR223_CETN3_WT

82961

Gateway Donor vector containing CETN3, part of the Target Accelerator Plasmid Collection.

Berger et al Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/162656/49046a75-acd3-4515-9022-9d511cf36bb6/addgene-plasmid-82961-sequence-162656.gbk

full

LOCUS sequence-162656- 3291 bp DNA circular SYN 26-AUG-2024 DEFINITION Gateway Donor vector containing CETN3, part of the Target Accelerator Plasmid Collection. . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3291) AUTHORS Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS TITLE High-throughput Phenotyping of Lung Cancer Somatic Mutations. JOURNAL Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28. PUBMED 27478040 REFERENCE 2 (bases 1 to 3291) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28" FEATURES Location/Qualifiers source 1..3291 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 43..65 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 57..74 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 58..74 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 90..189 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(691..790) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(807..826) /label=T7 /note="T7 promoter, forward primer" promoter complement(808..826) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(831..847) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(831..847) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1278..2069 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 2162..2750 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2651..2670 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2904..2921 /label=L4440 /note="L4440 vector, forward primer" terminator complement(3080..3107) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(3199..3285) /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta 61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga 121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa 181 aagttggcat gagtttagct ctgagaagtg agcttgtagt ggacaaaaca aagaggaaaa 241 aaagaagaga actgtctgag gaacagaaac aagaaattaa agatgctttt gaactatttg 301 atacagacaa agatgaagca atagattatc atgaattaaa ggtggcaatg agagccttgg 361 ggtttgatgt aaaaaaagct gatgtactga agattcttaa agattatgac agagaagcca 421 cagggaaaat cacctttgaa gattttaatg aagttgtgac agactggata ttggaaagag 481 atccccatga agaaatactc aaggcattta aactatttga tgatgatgat tcaggtaaaa 541 taagcttgag gaatttgcga cgtgttgcta gagaattggg tgaaaacatg agtgatgaag 601 aacttcgagc tatgatagaa gaatttgaca aagatggtga tggagaaata aaccaagagg 661 agttcattgc tattatgact ggtgacattt aaccaacttt cttgtacaaa gttggcatta 721 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa 781 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg 841 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa 901 atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac 961 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat 1021 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca 1081 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg 1141 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg 1201 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta 1261 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag 1321 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg 1381 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga 1441 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg 1501 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg 1561 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca 1621 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc 1681 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg 1741 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact 1801 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt 1861 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc 1921 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag 1981 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag 2041 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta 2101 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 2161 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 2221 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 2281 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 2341 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 2401 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 2461 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 2521 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 2581 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 2641 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 2701 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 2761 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 2821 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 2881 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa 2941 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt 3001 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg 3061 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg 3121 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt 3181 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat 3241 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc t //

RCAS(A) mPea3 EnR (CT#1120)

13909

Cliff Tabin

Brent et al Development. 2004 Aug . 131(16):3885-96.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/20132/a3e983ca-00a9-430d-9443-f2d4b22926ec/addgene-plasmid-13909-sequence-20132.gbk

partial

LOCUS sequence-20132-e 756 bp DNA linear UNA 25-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 756) AUTHORS Brent AE, Tabin CJ TITLE FGF acts directly on the somitic tendon progenitors through the Ets transcription factors Pea3 and Erm to regulate scleraxis expression. JOURNAL Development. 2004 Aug . 131(16):3885-96. PUBMED 15253939 REFERENCE 2 (bases 1 to 756) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Development. 2004 Aug . 131(16):3885-96." FEATURES Location/Qualifiers source 1..756 /mol_type="genomic DNA" /organism="unspecified" ORIGIN 1 ggaaagaagc tattggtaat tgtgaaatac gcttttgtct gtgtgctgca ggagctgagc 61 tgactctgct ggtggcctcg cgtaccactg tggcatcgat tctagaccac tgtggccagg 121 cggtagctgg gacgtgcagc cgaccaccat gcggggtgcc ttacaactgt ggcagtttct 181 ggtggccctg ctggatgacc caacaaatgc tcatttcatt gcttggacag gccggggaat 241 ggagtttaaa ctaattgaac ctgaagaggt tgccaggctc tggggtatcc agaagaaccg 301 gccagccatg aattatgaca agctgagccg ctcgctgcga tactattatg agaaaggcat 361 catgcagaag gtggctggcg aacgctacgt gtacaagttt gtgtgcgagc cggaggccct 421 gttctctctg gccttcccag ataatcaacg tccagctctg aaggctgagt ttgaccggcc 481 agtcagtgag gaggacacag tccctttgtc ccacttggat gagagtcctg cctacctccc 541 agaactcact ggccccgctc cgcccttcgg ccacagaggt ggatattctt acgaattccg 601 cgatgccctg gaggatcgct gcagcccaca gtcagcgccc agccccatta ccctacaaat 661 gcagcatctt caccaccagc aacagcagca gcagcaacag cagcagcaaa tgcagcacct 721 ccaccagctg cagcaactgc agcagttgca ccaaca //

Anti-CDY1/2 [N143/38R]

206535

Mammalian Expression Plasmid of anti-CDY1/2 (Human). Derived from hybridoma N143/38.

James Trimmer

Mitchell et al Sci Rep. 2023 Sep 27;13(1):16200. doi: 10.1038/s41598-023-43233-4.

partial

JPF0512

124003

Encodes promoter-less expression of mAzamiGreen with an mScarlet normalization in a PiggyBac destination vector

Hana El-Samad

Fonseca et al ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 10.1021/acssynbio.9b00322. Epub 2019 Nov 5.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/252361/4ac48f2a-4dcf-45d7-bdfe-14b548fa5bb4/addgene-plasmid-124003-sequence-252361.gbk

full

LOCUS sequence-252361- 9359 bp DNA circular SYN 26-AUG-2024 DEFINITION Encodes promoter-less expression of mAzamiGreen with an mScarlet normalization in a PiggyBac destination vector. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9359) AUTHORS Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H TITLE A Toolkit for Rapid Modular Construction of Biological Circuits in Mammalian Cells JOURNAL ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 10.1021/acssynbio.9b00322. Epub 2019 Nov 5. PUBMED 31686495 REFERENCE 2 (bases 1 to 9359) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606" FEATURES Location/Qualifiers source 1..9359 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 540..743 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 694..714 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 765..774 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 771..1799 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" CDS 771..1799 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" polyA_signal 1817..2024 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 2231..2293 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" promoter complement(2344..2362) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS complement(2402..3217) /codon_start=1 /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" CDS complement(2402..3217) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 3125..3144 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3209..3228 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3442..4030 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3931..3950 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4142..4161 /label=T7 /note="T7 promoter, forward primer" promoter 4142..4160 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" repeat_region 4190..4224 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" CDS 4765..4785 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4801..5472 /codon_start=1 /product="humanized monomeric Azami-Green fluorescent protein" /label=hmAzami-Green /note="mammalian codon-optimized" /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS QA" CDS 4801..5472 /codon_start=1 /product="monomeric Azami-Green fluorescent protein (Karasawa et al., 2003)" /label=mAzami-Green /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS QA" polyA_signal 5499..5723 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 5971..6274 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 6547..7563 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 7487..7509 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 7571..7590 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 7624..8001 /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSAK" primer_bind complement(8875..8894) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 8940..8995 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(8994..9013) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" ORIGIN 1 gttacagaga tgttacgaac cctgaacagg ttgactatta attgtgtctg cttgatgtgg 61 acaccaggtg gcgctggaca tcagatttgg agaggcagtt gtctagggaa ccgggctctg 121 tgccagcgca ggaggcaggc tggctctcct attccaggga tgctcatcca ggaaggaaag 181 gttgcatgct ggacacacta accttacagc ggtgggctct atggggtacc tacgtaggta 241 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 301 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg 361 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta 421 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga 481 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg 541 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 601 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 661 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 721 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatagccacc atgggtaaaa 781 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtgt 841 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag 901 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg 961 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat 1021 ttagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc 1081 tgcctgaaac cgaactgccc gctgttctgc aaccggtcgc ggaggccatg gatgcgatcg 1141 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc 1201 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc 1261 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc 1321 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca 1381 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg 1441 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg 1501 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc 1561 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt 1621 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga 1681 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag 1741 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataac 1801 tcgagcgacg cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 1861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 1921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 1981 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2041 agtatcgata ctagttaaaa gttttgttac tttatagaag aaattttgag tttttgtttt 2101 tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag 2161 tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg atatacagac 2221 cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc acaatatgat 2281 tatctttcta gggttaatct agtatacgcg tatgcggccg cttaattaat ccagcttttg 2341 ttccctttag tgagggttaa ttccgagcgg ccgcgtgtta caaccaatta accaattctg 2401 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa 2461 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc 2521 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac 2581 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga 2641 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc 2701 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt 2761 gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg 2821 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat 2881 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat 2941 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt 3001 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa 3061 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga 3121 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg 3181 gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta 3241 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc 3301 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg 3361 atcagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 3421 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 3481 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 3541 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 3601 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 3661 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 3721 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 3781 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 3841 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 3901 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 3961 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 4021 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 4081 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg cggccgccaa 4141 ttaatacgac tcactatagg gcgaattggg gcgcgccatt ctagattaac cctagaaaga 4201 tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt ctaaatagcg 4261 cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc cgtgaggcgt 4321 gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg agtcaaaatg 4381 acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat tatattgtta 4441 tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta tagatatcaa 4501 ctagaatgct agcatgggcc cagatctcca tgggccctga attcgcatct agactgaact 4561 ggccgataat tgcagacgag gagcatcgcc cttccccggc cctcaggtaa gaggaccaaa 4621 taccgtagcc gtttccaatt tcagtccttt agcgccacct ggtgctaact actctatcac 4681 gcttttatcc aataactacc tttgtaaatt tcctttcaaa agttctggcc gggcgcggtg 4741 gcaacgggcg gaggaggtag tatgcccaag aagaagagga aggtggagga cgccggttct 4801 gtgagcgtga tcaagcccga gatgaagatc aagctgtgca tgaggggcac cgtgaacggc 4861 cacaacttcg tgatcgaggg cgagggcaag ggcaacccct acgagggcac ccagatcctg 4921 gacctgaacg tgaccgaggg cgcccccctg cccttcgcct acgacatcct gaccaccgtg 4981 ttccagtacg gcaacagggc cttcaccaag taccccgccg acatccagga ctacttcaag 5041 cagaccttcc ccgagggcta ccactgggag aggagcatga cctacgagga ccagggcatc 5101 tgcaccgcca ccagcaacat cagcatgagg ggcgactgct tcttctacga catcaggttc 5161 gacggcacca acttcccccc caacggcccc gtgatgcaga agaagaccct gaagtgggag 5221 cccagcaccg agaagatgta cgtggaggac ggcgtgctga agggcgacgt gaacatgagg 5281 ctgctgctgg agggcggcgg ccactacagg tgcgacttca agaccaccta caaggccaag 5341 aaggaggtga ggctgcccga cgcccacaag atcgaccaca ggatcgagat cctgaagcac 5401 gacaaggact acaacaaggt gaagctgtac gagaacgccg tggccaggta ctccatgctg 5461 cccagccagg ccggatccta actcgagcga cgcctcgact gtgccttcta gttgccagcc 5521 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 5581 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 5641 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 5701 tggggatgcg gtgggctcta tgggctggtt ctgtatgggc acagacaacc taactaattt 5761 cctggccctc aaggtgactt gcttgaactt gccaccttcg ccactagggg gcagcattgg 5821 tttacacagg gtgaacaaac gttcaattct aaactaatat tctttggtgg gaaagtgtgt 5881 tcattttggt ttgttttgaa gaagtcactt catacctttg aactggggtt aagctggtta 5941 actcccaaaa acgccatata tggagttccg cgttacataa cttacggtaa atggcccgcc 6001 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt 6061 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca 6121 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg 6181 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca 6241 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt 6301 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt 6361 attttgtgca gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcgag 6421 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga 6481 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg 6541 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc 6601 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct 6661 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg 6721 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt 6781 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt 6841 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg 6901 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg 6961 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc 7021 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg 7081 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc 7141 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagt 7201 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag 7261 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc 7321 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc 7381 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc 7441 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg 7501 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta 7561 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcgcc 7621 actatgccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa gaaggcggtg 7681 actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga gagctattcc 7741 atctatgtgt acaaggttct gaagcaggtc caccctgaca ccggcatttc gtccaaggcc 7801 atgggcatca tgaattcgtt tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc 7861 cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca gacggccgtg 7921 cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac taaggccatc 7981 accaagtaca ccagcgctaa gggttctgta tcaaaaggcg aggcggtcat taaagagttc 8041 atgaggttta aagtccatat ggaaggaagc atgaatggtc acgagtttga gatagaggga 8101 gagggtgagg gaaggccgta cgaagggacg caaactgcga agttgaaggt aacgaaaggc 8161 ggccctctgc cattctcttg ggacatcctt tcaccgcaat ttatgtacgg ctctagggcc 8221 ttcacaaaac accccgccga tattccggat tactataaac aaagttttcc tgagggcttc 8281 aagtgggaaa gagtaatgaa cttcgaggat gggggagctg tgacggttac tcaagacact 8341 agcctggagg acggtactct tatatacaag gtaaaactaa gagggacgaa ttttccgcca 8401 gacggccccg tgatgcaaaa aaaaactatg gggtgggaag catccaccga gaggctgtac 8461 cccgaagacg gggtcctgaa gggtgatatt aaaatggcat taagattaaa agatggcggg 8521 agatacttag ctgacttcaa gactacatat aaagccaaaa aacctgtaca gatgcctgga 8581 gcttacaatg tggatcgtaa gctggatatt acatcccaca acgaagatta taccgtggtg 8641 gagcagtacg agcgttcaga ggggcgtcac tccaccgggg gtatggatga attatacaag 8701 agcggtctta gaagcagggc gcaagcgtca aattcagcgg tagatggaac agcaggtcca 8761 ggtagtactg gcagcagagg atcctaactc gagattcact cctcaggtgc aggctgccta 8821 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt 8881 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta 8941 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg 9001 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt 9061 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca 9121 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg 9181 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc 9241 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt 9301 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tgctgagca //

pMx/ATP8B3-HA

209228

Mammalian expression of ATP8B3

Hye-Won Shin

Takatsu et al J Biol Chem. 2014 Nov 28;289(48):33543-56. doi: 10.1074/jbc.M114.593012. Epub 2014 Oct 14.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/412997/dabbc8f5-7033-4efd-92c7-d1810d244c1e/addgene-plasmid-209228-sequence-412997.gbk

full

LOCUS sequence-412997- 10338 bp DNA circular SYN 27-AUG-2024 DEFINITION Mammalian expression of ATP8B3. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10338) AUTHORS Takatsu H, Tanaka G, Segawa K, Suzuki J, Nagata S, Nakayama K, Shin HW TITLE Phospholipid flippase activities and substrate specificities of human type IV P-type ATPases localized to the plasma membrane. JOURNAL J Biol Chem. 2014 Nov 28;289(48):33543-56. doi: 10.1074/jbc.M114.593012. Epub 2014 Oct 14. PUBMED 25315773 REFERENCE 2 (bases 1 to 10338) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1074/jbc.M114.593012"; journalName: "J Biol Chem"; date: "2014-11-28- 28"; volume: "289"; issue: "48"; pages: "33543-56" FEATURES Location/Qualifiers source 1..10338 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 22..615 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 678..1035 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1094..1510 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1440..1462 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1478..1494 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 1520..1894 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind 1832..1848 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" protein_bind 1938..1962 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(5914..5938) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 5970..5996 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(6165..6185) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6170..6499 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6350..6485 /label=SV40 ori /note="SV40 origin of replication" CDS 6856..7650 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6910..6929) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7520..7539 /label=Neo-F /note="Neomycin resistance gene, forward primer" LTR 7843..8312 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(8442..8459) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8613..9201) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8693..8712) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9365..10225) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 9988..10007 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10226..10330) /gene="bla" /label=AmpR promoter ORIGIN 1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta 61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc 121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag 181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat 241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg 301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct 361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg 421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc 481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc 541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag 601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg 661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg 721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg 781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc 841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg 901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct 961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc 1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt 1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa 1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac 1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa 1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc 1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt 1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc 1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc 1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa 1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca 1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca 1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg 1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac 1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc 1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat 1921 ctctagttaa gctatcaaca agtttgtaca aaaaagcagg ctctttaaag gaaccaattc 1981 agtcgactgg atccaccatg ggcactggcc ccgctcagac tcccaggagc accagagctg 2041 gccctgagcc aagccctgcc ccaccaggac ctggggacac gggtgactca gacgtgactc 2101 aggaaggctc aggtcctgct ggcatccgcg gaggtgagac ggtgatcaga gctgggatgg 2161 gagactcccc aggcagaggg gcacccgaga ggaggcacaa ggcccagcct ggccgggcta 2221 ggaagtatga atggagacca gaaggcccca ccagcatggg cagcctcggc cagagagaag 2281 atctccaaga tgaggacagg aactcagcat tcacctggaa ggtccaggcc aacaaccgtg 2341 cctacaacgg gcagttcaag gagaaggtga tcctgtgctg gcaaaggaag aaatacaaga 2401 ccaatgtcat ccgcacggcc aagtacaact tctactcgtt cctgccgctg aacctgtacg 2461 agcagttcca ccgcgtgtcc aacctgttct tcctcatcat catcatcctg cagagcattc 2521 ccgacatctc cacgctgccc tggttctcgc tcagtacccc tatggtctgc ctcctcttca 2581 tccgtgccac ccgggacctg gtggacgaca tggggagaca caagagtgac agagccatca 2641 acaacagacc ctgccagatt ctgatgggga agagcttcaa gcagaagaaa tggcaggatc 2701 tgtgcgtggg ggatgtggtc tgtctccgca aggacaacat cgtcccagcc gacatgctct 2761 tgctggccag cacggagccc agcagcctgt gctatgtgga gacggtggac attgacgggg 2821 agaccaactt gaagttcaga caggccctga tggtcaccca caaagaactg gccactataa 2881 agaagatggc gtcctttcaa ggcacagtga cgtgtgaggc gcctaacagt cggatgcacc 2941 acttcgtggg gtgcctggaa tggaatgaca agaaatactc cctggacatt ggcaacctcc 3001 tcctccgagg ctgcaggatt cgcaacacag acacctgcta tggactggtc atttatgctg 3061 gttttgacac aaaaattatg aagaactgtg gcaagatcca tttgaagaga accaagctgg 3121 acctcctgat gaacaagctg gtggttgtga tcttcatctc cgtggtgctt gtctgcctgg 3181 tgttggcctt cggcttcggt ttctcagtca aagaattcaa agaccaccac tactacctct 3241 cgggggtgca tgggagcagc gtggccgcag agtccttctt cgtcttctgg agcttcctca 3301 tcctgctcag cgtcaccatc ccgatgtcca tgttcatcct gtccgagttc atctacctgg 3361 ggaacagcgt cttcatcgac tgggacgtgc agatgtacta caagccgcag gacgtgcctg 3421 ccaaggcccg cagcaccagc ctcaacgacc acctgggcca ggtggaatac atcttctcgg 3481 acaagacggg cacgctcacg cagaacatct tgaccttcaa caagtgctgc atcagcggcc 3541 gcgtctatgg gccggattca gaggccacga cccgacctaa ggagaacccc tacctctgga 3601 acaagttcgc cgacgggaag ctgctcttcc acaatgcggc cctgctgcac ctcgtgcgga 3661 ccaacgggga cgaggccgtg cgggagttct ggcgcctgct ggccatctgc cacacggtga 3721 tggtgcggga gagcccccgt gagcgcccag accagctgtt gtaccaggcg gcctcccccg 3781 acgagggggc gctggtcacc gcagcccgga acttcggcta cgtgttcctg tcccgcaccc 3841 aggacaccgt cacgatcatg gagctggggg aggaacgggt ctaccaggtc ctggccataa 3901 tggacttcaa cagcacgcgc aaacggatgt cggtgctggt tcgaaagcca gagggcgcca 3961 tctgcctgta caccaagggc gccgacacgg tcatcttcga acgcttgcac aggagggggg 4021 caatggaatt tgccacagag gaggccttgg ctgcctttgc ccaggagacc ctgcggacac 4081 tgtgcctggc ctacagggag gtggctgagg acatttacga ggactggcag cagcgccacc 4141 aggaggccag cctcctgctg cagaaccggg cacaggccct gcaacagctg ctgggagcca 4201 cagccatcga ggacagactc caggacggtg tccctgaaac catcaaatgt ctcaagaaga 4261 gcaacatcaa aatatgggtg ctcaccgggg acaagcagga aacggctgtg aacatcggct 4321 tcgcctgcga gctgctgtca gagaatatgc tcattctgga ggagaaggag attagccgca 4381 tcctggagac ctactgggaa aacagtaaca accttctaac cagggagtcc ctgtcgcagg 4441 tcaagctggc cttggtcatt aacggagact tcctggacaa actgctggtg tccctgcgga 4501 aggagccgcg cgccctggcg cagaacgtga acatggacga ggcgtggcag gagctcggcc 4561 agtccaggag ggatttcctc tacgccaggc gcctgtccct gctgtgccgg aggttcgggc 4621 tcccgctggc tgcaccgcca gcccaggact ccagagcccg ccgtagctcc gaggtgctgc 4681 aggagcgcgc cttcgtggac ctggcgtcca agtgccaggc ggtcatctgc tgccgcgtga 4741 cgcccaagca gaaggccctg atcgtggccc tggtcaagaa gtaccaccag gtggtgaccc 4801 tggccatcgg ggacggtgcc aacgacatca acatgatcaa gaccgcggac gtgggcgtgg 4861 ggctggcggg ccaggagggc atgcaggcag ttcagaacag cgacttcgtg ctcggccagt 4921 tctgcttcct gcagcgcctc ctgctggtgc acggccgctg gtcctacgtg cggatctgca 4981 agttcctgcg ctacttcttc tacaagagca tggccagcat gatggtgcag gtctggtttg 5041 cctgctacaa cggcttcacc ggccagcccc tgtatgaagg atggttcctg gctcttttca 5101 acctcctgta cagcaccctg ccagttctct acattgggct ctttgagcag gacgtgagcg 5161 cagagcagag cctggagaag ccggagctgt acgtggtggg gcagaaggac gagctcttca 5221 actactgggt cttcgtccaa gccatcgccc atggtgtgac cacctctctg gtcaacttct 5281 tcatgacact gtggatcagc cgcgacacgg cgggacccgc cagcttcagc gaccaccagt 5341 cctttgcggt cgtggtggcc ctgtcttgcc tgctgtccat caccatggag gtcattctta 5401 tcatcaagta ctggaccgcc ctgtgcgtgg cgaccatcct cctcagcctt ggtttctacg 5461 ccatcatgac taccaccacc cagagcttct ggctcttcag agtatccccc acgaccttcc 5521 cgtttctgta tgccgacctc agcgtgatgt cctctccctc catcctgctg gtggtcctgc 5581 tgagtgtgtc cataaacacc ttccctgtcc tggccctccg agtcatcttc ccagccctca 5641 aggagctacg tgccaaggag gagaaggtgg aggagggccc cagcgaggag attttcacca 5701 tggagccctt gcctcatgta caccgggagt ctcgtgcccg ccgttccagc tatgctttct 5761 cccaccgtga gggatatgca aacctcatca ctcagggcac aattctgcgg aggggaccag 5821 gggtcagcag tgacatagca tctgaatccc tagacccatc tgatgaagag gcagcttcga 5881 gcccaaaaga gtcacaggcc tcgagatatc tagacccagc tttcttgtac aaagtggttg 5941 atctagaggg cccgcggttc gaactcgagt atccatacga tgtaccagac tacgcagaga 6001 tccactagtt caattccagc tgagcgccgg tcgctaccat taccagttgg tctggtgtca 6061 aaaataataa taaccgggca ggccatgtct gcccgtattt cgcgtaagga aatccattat 6121 gtactattta aactcgagcg gccgccagca cagtggtcga ctgtggaatg tgtgtcagtt 6181 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 6241 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 6301 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 6361 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 6421 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 6481 aggcctaggc ttttgcaaaa agcttcacgc tgccgcaagc actcagggcg caagggctgc 6541 taaaggaagc ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat 6601 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct 6661 tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg 6721 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg 6781 gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga 6841 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 6901 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 6961 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 7021 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 7081 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 7141 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 7201 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 7261 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 7321 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 7381 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 7441 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 7501 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 7561 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 7621 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgca tcgtcgacga 7681 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta 7741 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga 7801 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca 7861 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg 7921 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa 7981 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc 8041 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg 8101 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc 8161 tcactcggcg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa 8221 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga 8281 gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg 8341 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc 8401 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 8461 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 8521 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 8581 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 8641 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 8701 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 8761 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 8821 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 8881 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 8941 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 9001 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 9061 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 9121 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 9181 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 9241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 9301 tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg 9361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 9421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 9481 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 9541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 9601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 9661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 9721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 9781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 9841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 9901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 9961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 10021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 10081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 10141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 10201 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 10261 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 10321 gggttccgcg cacatttc //

Sph1-VC

89756

expresses human Synphilin fused to the C-terminal part of Venus

Tiago Outeiro

Lázaro et al bioRxiv 2024.03.05.583522

https://media.addgene.org/snapgene-media/v2.0.0/sequences/178646/7a167a80-3b9d-4b4b-891d-0d1e05f52a62/addgene-plasmid-89756-sequence-178646.gbk

full

LOCUS sequence-178646- 8363 bp DNA circular SYN 26-AUG-2024 DEFINITION expresses human Synphilin fused to the C-terminal part of Venus. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8363) AUTHORS Lazaro DF, Amen T, Gerhardt E, Santos PI, Milovanovic D, Hoglinger G, Outeiro TF TITLE Synphilin-1 as a modulator of aSyn assembly JOURNAL bioRxiv 2024.03.05.583522 REFERENCE 2 (bases 1 to 8363) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 2024.03.05.583522" FEATURES Location/Qualifiers source 1..8363 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 11..32 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(151..171) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 156..485 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 336..471 /label=SV40 ori /note="SV40 origin of replication" primer_bind 398..417 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 552..1346 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(606..625) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1216..1235 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1520..1641 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1557..1576) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1611..1630 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1690..1706) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1690..1706) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1703..1725) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1714..1730 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(1738..1768) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1783..1804 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1921..1938) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2092..2680) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2172..2191) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2851..3711) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3474..3493 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3712..3816) /gene="bla" /label=AmpR promoter primer_bind complement(3891..3910) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 4082..4461 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4462..4665 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4616..4636 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4710..4729 /label=T7 /note="T7 promoter, forward primer" promoter 4710..4728 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 7703..7724 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(7801..7818) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 7807..8031 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin join(8077..8363,1..142) /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8164..8183) /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 61 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 121 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 421 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 541 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 601 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 661 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 721 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 781 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 841 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 901 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 961 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 1021 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 1081 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 1141 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 1201 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 1261 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 1321 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 1381 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 1441 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 1501 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 1561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 1621 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 1681 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 1741 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 1801 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 1861 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 1921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 1981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 2041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 2101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 2161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 2221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 2281 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 2341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 2401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 2461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 2521 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 2581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 2641 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 2701 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 2761 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 2821 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 2881 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 2941 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 3001 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 3061 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 3121 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 3181 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 3241 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 3301 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 3361 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 3421 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 3481 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 3541 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 3601 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 3661 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 3721 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 3781 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 3841 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc 3901 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag 3961 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag 4021 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt 4081 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc 4141 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 4201 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 4261 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 4321 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 4381 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 4441 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 4501 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 4561 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 4621 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga 4681 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct 4741 ggctagcgtt taaacttaag atggaagccc ctgaatacct tgatttggat gaaattgact 4801 ttagtgatga catatcttat tcagtcacat cactcaagac gatcccagaa ctgtgccgaa 4861 gatgtgatac gcaaaacgaa gacagatcag tttctagctc tagctggaat tgtggcatct 4921 caactcttat tacaaacacg caaaagccca caggaatcgc tgatgtgtac agtaagttcc 4981 gcccagtgaa gcgggtttcg ccactgaaac atcagccaga gactctggag aacaatgaaa 5041 gtgatgacca aaagaaccag aaagtggttg agtaccagaa agggggtgag tctgacctgg 5101 gcccccagcc tcaggagctt ggccctggag atggagtggg cggcccacca ggtaagagct 5161 ctgagcccag cacatcgctg ggtgaactgg agcactacga cctcgacatg gatgagattc 5221 tggatgtgcc ttatattaaa tccagtcagc agcttgcctc ttttaccaag gtgacttcag 5281 aaaaaagaat tttgggctta tgcacaacca tcaatggcct ttctggcaaa gcctgctcta 5341 caggaagttc tgagagctca tcatccaaca tggcaccatt ttgtgttctt tctcccgtga 5401 aaagccctca cttgagaaaa gcatcagctg tcatccacga ccagcacaag ctgtccactg 5461 aagaaaccga gatctcacct cctctggtta aatgtggctc tgcatatgag cctgaaaacc 5521 agagtaaaga cttcctaaac aagacattta gtgatcctca tggtcgaaaa gttgagaaga 5581 caacaccaga ctgccagctc agggccttcc acctacaatc ctcagcagca gaatccaaac 5641 cagaagagca ggtcagtggc ctaaaccgga ccagctccca aggcccagaa gaaaggagtg 5701 agtatctgaa aaaagtgaaa agcatcttga acattgttaa agaaggacag atctctctcc 5761 tgccacacct agctgcagac aatctagaca aaattcacga cgaaaatgga aacaatctat 5821 tacatattgc ggcgtcacag ggacacgcag agtgtctaca gcacctcact tctttgatgg 5881 gagaagactg cctcaatgag cgcaacactg agaagttgac tccagcaggc ctggccatta 5941 agaatggtca gttggagtgc gtacgctgga tggtgagcga aacagaagcc attgcagaac 6001 tgagttgttc taaggatttt ccaagcctta ttcattacgc aggttgctat ggccaggaaa 6061 agattcttct gtggcttctt cagtttatgc aagaacaggg catctcgttg gatgaagtag 6121 accaggatgg caacagtgcc gttcacgtag cctcacagca tggctacctt ggatgcatac 6181 agaccttggt tgaatatgga gcaaatgtca ccatgcagaa ccacgctggg gaaaagccct 6241 cccagagcgc cgagcggcag gggcacaccc tgtgctccag gtacctggtg gtggtggaga 6301 cctgcatgtc gctggcctct caagtggtga agttaaccaa gcagctaaag gaacaaacag 6361 tagaacgtgt cacgctgcag aaccaactcc aacaatttct agaagcccag aaatcagagg 6421 gcaagtcact cccttcttca cccagttcgc catcctcacc tgcctccaga aagtcccagt 6481 ggaaatctcc agatgcagat gatgattctg tagccaaaag caagccagga gtccaagagg 6541 ggattcaggt tcttggaagc ctgtcagcct ccagccgggc tagacccaaa gcaaaagatg 6601 aagattctga taaaatctta cgccagttat tgggaaagga aatctcagaa aatgtctgca 6661 cccaggaaaa actgtccttg gaattccagg atgctcaggc ttcctctaga aattctaaaa 6721 agatcccact ggagaagagg gaactgaagt tagccaggct gagacagctg atgcagaggt 6781 cactgagtga gtctgacaca gactccaaca actctgagga ccccaagact accccagtga 6841 ggaaggctga ccgaccaagg ccgcagccca ttgtagaaag cgtagagagt atggacagcg 6901 cagaaagcct gcacctgatg attaagaaac acaccttggc atcaggggga cgcaggtttc 6961 ctttcagcat caaggcctcc aaatccctgg atggccacag cccatctccc acctcagaga 7021 gcagcgaacc agacttagaa tcccagtatc caggctcagg gagtattcct ccaaaccagc 7081 cctctggtga ccctcagcag cccagccctg acagtactgc tgcccagaaa gttgccacaa 7141 gtcccaagag tgccctcaag tctccatctt ccaagcgtag gacatctcag aacttaaaac 7201 tgagagttac ctttgaggag cctgtggtgc agatggagca gcctagcctt gaactgaatg 7261 gagaaaaaga caaagataag ggaaggactc tccagcggac ctccacaagt aacgaatcgg 7321 gggatcaact gaaaaggcct tttggagcct ttcgatctat catggagaca ctaagtggca 7381 accaaaacaa taataataac taccaggcag ccaaccagct gaaaacctct acattgccct 7441 tgacctcact tgggaggaag acagatgcca agggaaaccc tgccagctcc gctagcaaag 7501 gaaagaataa ggcagcactc gagaagaacg gcatcaaggc caacttcaag atccgccaca 7561 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 7621 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag 7681 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 7741 ctctcggcat ggacgagctg tacaagtaat ctagagggcc cgtttaaacc cgctgatcag 7801 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 7861 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 7921 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 7981 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 8041 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 8101 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 8161 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 8221 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 8281 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 8341 gccctttgac gttggagtcc acg //

pDGB1_alpha1R

68226

GoldenBraid level 1 Alpha plant expression vector.

Diego Orzaez

Sarrion-Perdigones et al Plant Physiol. 2013 Jul;162(3):1618-31.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/135592/ffe554da-47be-464e-ae8d-c4cc56327a77/addgene-plasmid-68226-sequence-135592.gbk

full

LOCUS sequence-135592- 3196 bp DNA circular SYN 26-AUG-2024 DEFINITION GoldenBraid level 1 Alpha plant expression vector. . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3196) AUTHORS Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D TITLE GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. JOURNAL Plant Physiol. 2013 Jul;162(3):1618-31. PUBMED 23669743 REFERENCE 2 (bases 1 to 3196) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31" FEATURES Location/Qualifiers source 1..3196 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 87..108 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 123..153 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 161..177 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 166..188 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 185..201 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 185..201 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" misc_feature 214..270 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(271..288) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(271..287) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(280..302) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(496..515) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" misc_feature 635..659 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" rep_origin complement(750..1338) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(830..849) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1509..2324) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 2232..2251 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2316..2335 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2615..3050 /label=pSa ori /note="origin of replication from bacterial plasmid pSa" misc_feature join(3177..3196,1..3) /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA (truncated)" ORIGIN 1 taacggatcc gtctcaggag agcgagagac cccagctggc acgacaggtt tcccgactgg 61 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag 121 gctttacact atatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt 181 cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct 241 agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg 301 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg 361 gcgtaatagc gaagaggcct gcgccgatcg cccttcccaa cagttgcgca gcctgaatgg 421 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat 481 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 541 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 601 agctgtgggt ctctctccgt catgagacgg atcctgacag gatatattgg cgggtaaact 661 aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca aaaggccagg 721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 1021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 1081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 1141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 1201 ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa gagttggtag ctcttgatcc 1261 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 1321 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 1381 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 1441 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatgt gtaacattgg 1501 tctagtgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga 1561 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg 1621 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca 1681 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga 1741 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca 1801 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt 1861 cgtgattgcg cctgagcgag tcgaaatacg cgatcgctgt taaaaggaca attacaaaca 1921 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa 1981 tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt ggtgagtaac 2041 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc 2101 agccagttta gtctgaccat ctcatctgta acaacattgg caacgctacc tttgccatgt 2161 ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt cgcacctgat 2221 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt 2281 aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta 2341 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg 2401 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag 2461 ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag 2521 ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact 2581 ttctggctgg atgatggggc gattcaggcg atccccatcc aacagcccgc cgtcgagcgg 2641 gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa accgctaatg 2701 ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat ccacgtgaaa 2761 tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta ataaggtcac 2821 gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag atcaacagct 2881 tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa ttagcttttc 2941 aattatgaat atatatatca attattggtc gcccttggct tgtggacaat gcgctacgcg 3001 caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac 3061 aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa aaagaaaaag 3121 cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt agatcttggc 3181 aggatatatt gtggtg //

pET26b_Nb.AT110.i1

135481

E. coli periplasmic expression of high-affinity conformational nanobody Nb.AT110.i1 recognizing angiotensin II type 1 receptor

Andrew Kruse

Wingler et al Cell. 2019 Jan 24;176(3):479-490.e12. doi: 10.1016/j.cell.2018.12.006. Epub 2019 Jan 10.

partial

pmTurquoise2-H2A

36207

High

In vivo visualization of the nucleus (can be used for colocalization studies)

Dorus Gadella

Goedhart et al Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/44976/8ddfed9f-7412-4fe3-8761-1c2703046cc5/addgene-plasmid-36207-sequence-44976.gbk

full

LOCUS sequence-44976-e 5099 bp DNA circular SYN 25-AUG-2024 DEFINITION In vivo visualization of the nucleus (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5099) AUTHORS Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A TITLE Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. JOURNAL Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. PUBMED 22434194 REFERENCE 2 (bases 1 to 5099) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun."; date: "2012-03-20"; volume: "3:751. doi"; pages: " 10.1038/ncomms1738" FEATURES Location/Qualifiers source 1..5099 /mol_type="other DNA" /organism="synthetic DNA construct" polyA_signal 565..686 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(602..621) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 656..675 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(693..1148) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(830..851) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1042..1061 /label=F1ori-R /note="F1 origin, reverse primer" promoter 1175..1279 /gene="bla" /label=AmpR promoter promoter 1281..1638 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 1281..1472 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(1304..1324) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1489..1624 /label=SV40 ori /note="SV40 origin of replication" CDS 1673..2467 /codon_start=1 /product="aminoglycoside phosphotransferase" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS 1673..2467 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1727..1746) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2337..2356 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(2655..2674) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 2699..2746 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 3075..3663 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3564..3583 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 3838..4141 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4142..4345 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4296..4316 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 4384..4393 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS join(4390..5099,1..7) /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4435..4456) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4696..4715) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5043..5064 /label=EGFP-C /note="EGFP, forward primer" ORIGIN 1 gtacaagtac tcagatctcg agctcaagct tcgaattctg atgtcgggac gcggcaagca 61 gggaggcaaa gctcgcgcca aagccaagac ccgctcttct cgtgccggtc tccagttccc 121 cgtgggccga gtgcaccgac tgctccgcaa gggcaactat gctgagcggg tcggggccgg 181 cgcgccggtg tacctggcgg cggtgctgga gtacctgact gccgagatcc tggagctggc 241 gggcaacgcc gcccgcgaca acaagaagac ccgcattatc ccgcgccact tgcagctggc 301 catccgcaac gacgaggagc tcaacaagct gctgggcaaa gtaaccatcg ctcagggtgg 361 tgtcctgccc aacatccagg ctgtgctact gcccaagaag accgagagtc accacaaggc 421 caaaggcaaa taacgggatc caccggatct agataactga tcataatcag ccataccaca 481 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat 541 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa 601 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 661 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa 721 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 781 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 841 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 901 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg 961 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc 1021 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc 1081 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca 1141 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 1201 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 1261 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta 1321 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 1381 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 1441 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 1501 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 1561 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 1621 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga 1681 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 1741 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 1801 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga 1861 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 1921 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 1981 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 2041 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 2101 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 2161 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga 2221 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 2281 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 2341 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 2401 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 2461 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 2521 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 2581 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct 2641 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg 2701 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt 2761 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2821 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 2881 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt 2941 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 3001 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 3061 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 3121 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 3181 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 3241 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 3301 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 3361 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 3421 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 3481 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 3541 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 3601 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 3661 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 3721 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag 3781 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt 3841 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 3901 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg 3961 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 4021 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat 4081 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat 4141 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt 4201 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga 4261 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg 4321 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg 4381 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc 4441 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat 4501 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc 4561 tggcccaccc tcgtgaccac cctgtcctgg ggcgtgcagt gcttcgcccg ctaccccgac 4621 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc 4681 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc 4741 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc 4801 ctggggcaca agctggagta caactacttt agcgacaacg tctatatcac cgccgacaag 4861 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg 4921 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 4981 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat 5041 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagct //

pKDR-Lenti-TetOn

183555

(Empty Backbone)pKDR compatible TetOn lentiviral transfer plasmid

Erik Dent

Millette et al Front Genome Ed. 2022 Mar 28;4:803375. doi: 10.3389/fgeed.2022.803375. eCollection 2022.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/365362/d7c6a6cc-8553-4eb3-a5ae-fb205ec49d66/addgene-plasmid-183555-sequence-365362.gbk

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LOCUS sequence-365362- 10836 bp DNA circular SYN 25-AUG-2024 DEFINITION pKDR compatible TetOn lentiviral transfer plasmid. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10836) AUTHORS Millette MM, Holland ED, Tenpas TJ, Dent EW TITLE A Single Transcript Knockdown-Replacement Strategy Employing 5' UTR Secondary Structures to Precisely Titrate Rescue Protein Translation. JOURNAL Front Genome Ed. 2022 Mar 28;4:803375. doi: 10.3389/fgeed.2022.803375. eCollection 2022. PUBMED 35419562 REFERENCE 2 (bases 1 to 10836) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Front Genome Ed."; date: "2022-03-28"; volume: "4:803375. doi"; pages: " 10.3389/fgeed.2022.803375. eCollection 2022" FEATURES Location/Qualifiers source 1..10836 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 1..22 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 37..67 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 75..91 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 80..102 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 99..115 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 99..115 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 134..154 /label=T3 /note="T3 promoter, forward primer" promoter 136..154 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 182..408 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 409..589 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 636..761 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1254..1487 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1672..1716 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 1983..2100 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2146..2460 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 2154..2172 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2190..2208 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2225..2243 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2261..2279 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2297..2315 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2332..2350 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2368..2386 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 2433..2457 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 2486..2495 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 2492..3211 /codon_start=1 /product="bright green-yellow fluorescent protein derived from mClover2, with improved photostability (Bajar et al., 2003)" /label=mClover3 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" primer_bind complement(2537..2558) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 3145..3166 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3242..3828 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3409..3426) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3636..3655 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" ncRNA 4752..4766 /label=miR-30a loop /note="loop from the the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" misc_feature 4936..5524 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4989..5009) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5407..5418) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter 5537..6047 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 5966..5984 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 6067..6663 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6067..6086) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6563..6583 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 6664..6717 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 6718..7464 /codon_start=1 /product="improved tetracycline-controlled transactivator" /label=rtTA-Advanced /note="In the Tet-On(R) system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline." /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" LTR 7536..7769 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7841..7962 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7878..7897) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7932..7951 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 8002..8137 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(8157..8176) /label=T7 /note="T7 promoter, forward primer" promoter complement(8158..8176) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8186..8203) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8186..8202) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8195..8217) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8344..8799 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8431..8450) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8641..8662 /label=F1ori-F /note="F1 origin, forward primer" promoter 8825..8929 /gene="bla" /label=AmpR promoter CDS 8930..9790 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(9148..9167) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9961..10549 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10450..10469 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10703..10720 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga 121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta 181 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat 241 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat 301 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg 361 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg 421 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct 481 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 541 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga 601 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg 661 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga 721 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa 781 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa 841 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag 901 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat 961 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga 1021 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta 1081 agaccaccgc acagcaagct gccgctgatc ttcagacctg gaggaggaga tatgagggac 1141 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca 1201 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct 1261 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg 1321 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg 1381 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag 1441 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt 1501 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa 1561 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat 1621 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa 1681 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat 1741 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata 1801 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt 1861 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt 1921 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta 1981 acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata 2041 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt 2101 atcgatcacg agactagccg tcgactggcc gcgaatcgat atgtcgagtt tactccctat 2161 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt 2221 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa 2281 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca 2341 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt 2401 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc 2461 tggagaaacc ggttatactg cggccgccac catggtgagc aagggcgagg agctgttcac 2521 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt 2581 ccgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac 2641 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcgtggc 2701 ctgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc 2761 cgaaggctac gtccaggagc gcaccatctc tttcaaggac gacggtacct acaagacccg 2821 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga 2881 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca acagccacta 2941 cgtctatatc acggccgaca agcagaagaa ctgcatcaag gctaacttca agatccgcca 3001 caacgttgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg 3061 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc catcagtcca agctgagcaa 3121 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat 3181 tacacatggc atggacgagc tgtacaagta gcctgcaggg tcacctaaat aattccgccc 3241 ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt 3301 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc 3361 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag 3421 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac 3481 aaacaacctc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc 3541 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc 3601 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca 3661 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt 3721 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg 3781 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat gggctgcacc 3841 gtgagcgccg aggacaaggc ggcggccgag cgcggctctg gttccggcac tagtgtgagt 3901 aaaggagagg ccgttataaa agagttcatg cggttcaagg tacacatgga gggatcaatg 3961 aacggacacg agttcgaaat cgaaggggaa ggagagggca ggccctatga gggtactcag 4021 accgccaaac tcaaagtgac aaagggaggc cctttgcctt tctcttggga catcttgtca 4081 ccacaattta tgtatggctc ccgagcattt ataaaacacc cagcagacat acccgattac 4141 tacaagcagt ctttcccaga aggttttaaa tgggaaaggg tcatgaactt tgaggatggt 4201 ggtgctgtga cggtgacgca ggatacttct ttggaggacg gaacactcat ctacaaggta 4261 aagctcaggg gcacgaactt cccgccggac ggaccagtca tgcagaaaaa gactatggga 4321 tgggaggctt ccaccgaacg gttgtatccc gaggacggtg ttttgaaggg agatataaaa 4381 atggctcttc ggctgaaaga tggaggcaga tacttggcag attttaagac cacgtataag 4441 gcaaaaaaac cagtgcaaat gccgggagca tataacgttg accgcaaact ggacatcacc 4501 tcacataacg aggattatac agtagtagaa cagtacgaaa gaagcgaagg gaggcacagc 4561 accgggggta tggatgagct gtataagtga taggcatgct gtttgaatga ggcttcagta 4621 ctttacagaa tcgttgcctg cacatcttgg aaacacttgc tgggattact tcgacttctt 4681 aacccaacag aaggctcgag aaggtatatt gctgttgaca gtgagcgcac tctttctagg 4741 aggttgtgat agtgaagcca cagatgtatc acaacctcct agaaagagta tgcctactgc 4801 ctcggacttc aaggggctag aattcgagca attatcttgt ttactaaaac tgaatacctt 4861 gctatctctt tgatacattt ttacaaagct gaattaaaat ggtataaatt aaatcacttt 4921 tttcaattga cgcgtaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 4981 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 5041 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 5101 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 5161 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 5221 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 5281 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc 5341 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 5401 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 5461 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 5521 ccgcctgagc gcttccgggg ttggggttgc gccttttcca aggcagccct gggtttgcgc 5581 agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc 5641 gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc 5701 ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt 5761 gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc 5821 agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag 5881 ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag 5941 tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt 6001 cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagcaa ttcactgcag 6061 gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg 6121 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat 6181 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg 6241 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg 6301 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg 6361 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc 6421 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt 6481 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc 6541 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc 6601 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt 6661 gccgaaggta gaggttctct cctcacttgt ggtgatgttg aagaaaaccc tggtccaatg 6721 tctagactgg acaagagcaa agtcataaac ggagctctgg aattactcaa tggtgtcggt 6781 atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg 6841 tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac 6901 aggcatcata cccacttctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac 6961 aacgccaagt cataccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc 7021 ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt 7081 cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca 7141 ctgggctgcg tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct 7201 accaccgatt ctatgccccc acttctgaga caagcaattg agctgttcga ccggcaggga 7261 gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta 7321 aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt ttgacttaga catgctccca 7381 gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt 7441 gaccttgaca tgctccccgg gtaaggtacc tttaagacca atgacttaca aggcagctgt 7501 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctagctc actcccaacg 7561 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 7621 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 7681 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 7741 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 7801 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 7861 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 7921 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 7981 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 8041 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 8101 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct 8161 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa 8221 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 8281 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 8341 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 8401 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 8461 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 8521 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 8581 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 8641 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 8701 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 8761 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa 8821 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 8881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 8941 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 9001 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 9061 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 9121 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 9181 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 9241 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 9301 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 9361 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 9421 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 9481 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 9541 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 9601 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 9661 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 9721 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 9781 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 9841 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 9901 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 9961 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 10021 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 10081 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 10141 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 10201 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 10261 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 10321 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 10381 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 10441 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 10501 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 10561 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 10621 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 10681 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 10741 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt 10801 tcccgactgg aaagcgggca gtgagcgcaa cgcaat //

pGL0_89 [ORI_BC_14]

198990

Level 0 part

Gilbert et al bioRxiv 2023.05.19.541510

https://media.addgene.org/snapgene-media/v2.0.0/sequences/401237/f5782b9b-f6a6-4619-8282-5df4dacef070/addgene-plasmid-198990-sequence-401237.gbk

full

LOCUS sequence-401237- 2313 bp DNA circular SYN 27-AUG-2024 DEFINITION Level 0 part. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2313) AUTHORS Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z, Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N TITLE A scalable framework for high-throughput identification of functional origins of replication in non-model bacteria JOURNAL bioRxiv 2023.05.19.541510 REFERENCE 2 (bases 1 to 2313) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 2023.05.19.541510" FEATURES Location/Qualifiers source 1..2313 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(357..373) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(357..373) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(370..392) /label=M13/pUC Reverse /note="In lacZ gene" rep_origin complement(545..1133) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(625..644) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator complement(1266..1360) /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(1381..2040) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 1955..1974 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(2041..2143) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" primer_bind 2230..2252 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2244..2261 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2245..2261 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 ccggaatcta ttggcctatg tcactgaaac tatccaaaca ccccatgtcg atactgaacg 61 tatcgacgca tacctccttc cttgaaaacg cacaatcaag gtctcggtca gggtcacgcg 121 taggacgcgt cgagtagggt gacttgtggc cctttctttt gttccgcagc cacacgccgt 181 gtgaagctgg ggagtgagac cagtccttac ctgaagaata agtgtcagcc agtgtaaccc 241 gatgaggaac ccaaaaggcg aaccgggcca gacaacccgg cggtatcgca ctcaaagccg 301 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca 361 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat 421 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa 481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 1021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 1081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 1141 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc 1201 agcgtaaatg ggtagggggc ttcaaatcgt ccccccatac gatataagtt gttactagtg 1261 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga 1321 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa 1381 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac 1441 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc 1501 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc 1561 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt 1621 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga 1681 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt 1741 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc 1801 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg 1861 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat 1921 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg 1981 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat 2041 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct 2101 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attgatacct 2161 gaaacaaaac ccatcgtacg gccaaggaag tctccaataa ctgtgatcca ccacaagcgc 2221 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tcatgcataa tccgcacgca 2281 tctggaataa ggaagtgcca ttccgcctga cct //

p2xFlag CMV2-YAP2-2nd WW mutant

19047

Low

Marius Sudol

Oka et al J Biol Chem. 2008 Jul 17. ():.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/24945/98d1ed63-cdc8-433d-8213-33bb6dee82c0/addgene-plasmid-19047-sequence-24945.gbk

partial

LOCUS sequence-24945-e 861 bp DNA linear UNA 26-AUG-2024 DEFINITION natural linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 861) AUTHORS Oka T, Mazack V, Sudol M TITLE Mst2 and Lats kinases regulate apoptotic function of YAP. JOURNAL J Biol Chem. 2008 Jul 17. ():. PUBMED 18640976 REFERENCE 2 (bases 1 to 861) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Chem. 2008 Jul 17. ():." FEATURES Location/Qualifiers source 1..861 /mol_type="genomic DNA" /organism="unspecified" regulatory 93..102 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 102..125 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" ORIGIN 1 ttagcagagc tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg 61 acgatnagct tgcggccgcg aattcaagct tagccaccat ggactacaaa gacgatgacg 121 ataaagcaag gctcgaatcg gtacctaagg atcccgggca gcagccgccg cctcaaccgg 181 ccccccaggg ccaagggcag ccgccttcgc agcccccgca ggggcagggc ccgccgtccg 241 gacccgggca accggcaccc gcggcgaccc aggcggcgcc gcaggcaccc cccgccgggc 301 atcagatcgt gcacgtccgc ggggactcgg agaccgacct ggaggcgctc ttcaacgccg 361 tcatgaaccc caagacggcc aacgtgcccc agaccgtgcc catgaggctc cggaagctgc 421 ccgactcctt cttcaagccg ccggagccca aatcccactc ccgacaggcc agtactgatg 481 caggcactgc aggagccctg actccacagc atgttcgagc tcattcctct ccagcttctc 541 tgcagttggg agctgtttct cctgggacac tgacccccac tggagtagtc tctggcccag 601 cagctacacc cacagctcag catcttcgac agtcttcttt tgagatacct gatgatgtac 661 ctctgccagc aggttgggag atggcaaaga ntcttctggt cagagatact tcttaaatca 721 catcgatcag acacaacatg gcangacccn ggaaggcatg ctgtnntatg aanttaaacc 781 ccncntccnc atgcnnaaaa nganaaanat tttaaatcnt ttcgtntttn tnnnggtgga 841 anganatntt ttnngaantt a //

pLXV-EF1alpha-2xStrep-SARS-CoV-2-nsp14-IRES-Puro

141380

Medium

Lentiviral expression of SARS-CoV-2 protein; transient expression and generate lentivirus

Nevan Krogan

Gordon et al Nature. 2020 Jul;583(7816):459-468. doi: 10.1038/s41586-020-2286-9. Epub 2020 Apr 30.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/288093/c07d7a72-87ba-4911-9404-964b8966f340/addgene-plasmid-141380-sequence-288093.gbk

full

LOCUS sequence-288093- 10491 bp DNA circular SYN 25-AUG-2024 DEFINITION Lentiviral expression of SARS-CoV-2 protein; transient expression and generate lentivirus. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10491) AUTHORS Gordon DE, Jang GM, Bouhaddou M, Xu J, Obernier K, White KM, O'Meara MJ, Rezelj VV, Guo JZ, Swaney DL, Tummino TA, Huttenhain R, Kaake RM, Richards AL, Tutuncuoglu B, Foussard H, Batra J, Haas K, Modak M, Kim M, Haas P, Polacco BJ, Braberg H, Fabius JM, Eckhardt M, Soucheray M, Bennett MJ, Cakir M, McGregor MJ, Li Q, Meyer B, Roesch F, Vallet T, Mac Kain A, Miorin L, Moreno E, Naing ZZC, Zhou Y, Peng S, Shi Y, Zhang Z, Shen W, Kirby IT, Melnyk JE, Chorba JS, Lou K, Dai SA, Barrio-Hernandez I, Memon D, Hernandez-Armenta C, Lyu J, Mathy CJP, Perica T, Pilla KB, Ganesan SJ, Saltzberg DJ, Rakesh R, Liu X, Rosenthal SB, Calviello L, Venkataramanan S, Liboy-Lugo J, Lin Y, Huang XP, Liu Y, Wankowicz SA, Bohn M, Safari M, Ugur FS, Koh C, Savar NS, Tran QD, Shengjuler D, Fletcher SJ, O'Neal MC, Cai Y, Chang JCJ, Broadhurst DJ, Klippsten S, Sharp PP, Wenzell NA, Kuzuoglu-Ozturk D, Wang HY, Trenker R, Young JM, Cavero DA, Hiatt J, Roth TL, Rathore U, Subramanian A, Noack J, Hubert M, Stroud RM, Frankel AD, Rosenberg OS, Verba KA, Agard DA, Ott M, Emerman M, Jura N, von Zastrow M, Verdin E, Ashworth A, Schwartz O, d'Enfert C, Mukherjee S, Jacobson M, Malik HS, Fujimori DG, Ideker T, Craik CS, Floor SN, Fraser JS, Gross JD, Sali A, Roth BL, Ruggero D, Taunton J, Kortemme T, Beltrao P, Vignuzzi M, Garcia-Sastre A, Shokat KM, Shoichet BK, Krogan NJ TITLE A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. JOURNAL Nature. 2020 Jul;583(7816):459-468. doi: 10.1038/s41586-020-2286-9. Epub 2020 Apr 30. PUBMED 32353859 REFERENCE 2 (bases 1 to 10491) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/s41586-020-2286-9"; journalName: "Nature"; date: "2020-07"; volume: "583"; issue: "7816"; pages: "459-468" FEATURES Location/Qualifiers source 1..10491 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2337..3519 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2568..3510 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3467..3487 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 3553..3576 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 3613..3636 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 5241..5827 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(5408..5425) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5635..5654 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5847..6446 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(5847..5866) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6343..6363 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 6460..7048 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6513..6533) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6931..6942) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7255..7888 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(8016..8032) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8016..8032) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8029..8051) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8040..8056 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(8064..8094) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8109..8130 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8247..8264) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8418..9006) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8498..8517) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9177..10037) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 9800..9819 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10038..10142) /gene="bla" /label=AmpR promoter polyA_signal 10190..10324 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10227..10246) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10281..10300 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc 3541 gccgccacca tgtggtccca tccgcagttt gagaagggtg gtggttcagg cggaggctcc 3601 gggggcggga gctggtctca cccgcaattt gaaaaaggcg ctgcggctgc tgaaaatgta 3661 acgggcttgt ttaaagactg tagtaaagtg atcacaggac tccaccccac acaagcacct 3721 acccaccttt ccgtagatac gaagttcaaa acggaaggat tgtgtgtgga tataccaggg 3781 ataccaaagg atatgacgta ccgaaggctg atttccatga tgggttttaa gatgaattac 3841 caagttaatg gctaccccaa catgttcatc accagggagg aggcaattag acacgtaaga 3901 gcctggatag gcttcgatgt tgaaggttgc catgcaaccc gggaagccgt aggcacaaac 3961 cttccgttgc agcttggctt ttccacgggc gtcaacctcg ttgccgtacc gactggctat 4021 gttgacacgc cgaacaacac cgatttctct cgcgtaagtg ctaagcctcc tccgggagat 4081 caattcaagc atcttatacc tctcatgtac aagggactgc cgtggaatgt agttcgcatt 4141 aagatcgtgc agatgctctc cgatactttg aaaaatctga gcgacagggt ggtcttcgtt 4201 ctgtgggcac atggcttcga acttacttct atgaaatact ttgtaaaaat cgggcccgag 4261 cggacatgtt gtttgtgcga ccgacgagct acttgtttta gcacagcatc tgacacctac 4321 gcatgctggc accacagtat aggcttcgat tacgtctaca atccctttat gatagacgta 4381 caacaatggg gctttacggg taacttgcag agtaatcacg atctttactg ccaagttcat 4441 gggaacgcac acgtggcctc ctgcgatgcg ataatgacga ggtgcttggc cgtgcacgag 4501 tgctttgtta agcgggtcga ttggactata gagtatccca taatcggtga cgaacttaaa 4561 attaatgctg catgcaggaa agtgcagcac atggtagtaa aagcagcgct cctcgcggat 4621 aagtttcctg tgctccacga tatcggcaac cctaaggcca ttaaatgcgt cccccaagct 4681 gatgtagagt ggaaattcta tgacgctcag ccatgtagtg acaaagcata caagattgaa 4741 gaattgtttt attcatatgc gacacactcc gataagttca ctgacggagt ttgtctcttt 4801 tggaactgta atgttgatag gtatccggcg aatagcatag tgtgtcgatt tgacacgcgc 4861 gttctctcta atttgaacct ccctggctgc gacggggggt ctttgtacgt aaacaagcat 4921 gcatttcata ccccagcttt tgacaaatcc gctttcgtaa atctgaaaca acttcccttc 4981 ttctattatt cagatagccc ttgcgaatct cacggtaagc aagtggttag tgacattgac 5041 tatgtaccac ttaaaagtgc aacgtgcatt acacgatgca atttgggggg cgcggtttgc 5101 cgacaccatg ccaacgaata cagattgtac ctggatgcat ataacatgat gataagtgca 5161 gggtttagtc tttgggtgta taagcagttc gatacgtata atctctggaa cacatttacc 5221 cgattgcagt agggatcccg cccctctccc tccccccccc ctaacgttac tggccgaagc 5281 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct 5341 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt 5401 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct 5461 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc 5521 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag 5581 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc 5641 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga 5701 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt 5761 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc 5821 cacaacccac aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca 5881 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 5941 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 6001 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 6061 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 6121 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 6181 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 6241 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 6301 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 6361 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 6421 gcatgacccg caagcccggt gcctagacgc gtctggaaca atcaacctct ggattacaaa 6481 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 6541 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 6601 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 6661 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 6721 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 6781 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 6841 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 6901 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 6961 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 7021 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa 7081 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag 7141 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa 7201 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag 7261 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag 7321 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct 7381 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag 7441 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag 7501 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc 7561 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg 7621 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca 7681 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg 7741 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 7801 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 7861 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 7921 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta 7981 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 8041 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 8101 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 8161 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 8221 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 8281 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 8341 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 8401 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 8461 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 8521 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 8581 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 8641 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 8701 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 8761 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 8821 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 8881 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 8941 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 9001 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 9061 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 9121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 9181 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 9241 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 9301 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 9361 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 9421 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 9481 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 9541 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 9601 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 9661 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 9721 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 9781 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 9841 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 9901 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 9961 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 10021 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 10081 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 10141 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat 10201 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 10261 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 10321 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat 10381 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt 10441 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag t //

PP-CPUF2

25195

Bacterial expression for structure determination; may not be full ORF

Cheryl Arrowsmith

Structural Genomics Consortium Clones (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/31275/a59b2fd5-c865-4f76-add8-ced595136a8d/addgene-plasmid-25195-sequence-31275.gbk

partial

LOCUS sequence-31275-e 926 bp DNA linear UNA 26-AUG-2024 DEFINITION Bacterial expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 926) TITLE Structural Genomics Consortium Clones REFERENCE 2 (bases 1 to 926) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..926 /mol_type="genomic DNA" /organism="unspecified" CDS 61..78 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 88..105 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 384..401 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(454..472) /label=T7 Term /note="T7 terminator, reverse primer" terminator 468..515 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 552..910 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(639..658) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 849..870 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 gggtaattcc tctgaaatat tttgtttact ttaagaagga gatataccat gggcagcagc 61 catcatcatc atcatcacag cagcggcctg gttccgcgtg gtagttgcat acaaaaggtt 121 attgaagata agttaagttc agctttaaag cccacatttt tagaattagt agataaatct 181 tgtggatgtg gaacatcctt tgatgctgtc atagtttcta ataatttcga ggataaaaaa 241 ttattagata gacatagact tgttaacact atattaaaag aagaactaca aaatattcac 301 gccttttcta tgaaatgtca caccccactg gaatatgata aattaaaaag caaatgatga 361 cgaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca 421 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc 481 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat 541 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 601 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 661 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 721 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 781 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 841 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 901 ttttgattta taagggattt tgccga //

NEK8 gRNA (BRDN0001148289)

76942

3rd generation lentiviral gRNA plasmid targeting human NEK8

Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/189011/4e90c0ce-f524-428d-be53-32e699936671/addgene-plasmid-76942-sequence-189011.gbk

full

LOCUS sequence-189011- 8323 bp DNA circular SYN 26-AUG-2024 DEFINITION 3rd generation lentiviral gRNA plasmid targeting human NEK8. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8323) AUTHORS Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE TITLE Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. JOURNAL Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. PUBMED 26780180 REFERENCE 2 (bases 1 to 8323) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437." FEATURES Location/Qualifiers source 1..8323 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 87..1265 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 318..1256 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1213..1233 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1278..1877 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1278..1297) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1774..1794 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 1893..2481 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1946..1966) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2364..2375) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 2553..2786 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 2864..2985 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2901..2920) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2955..2974 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3025..3160 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(3180..3199) /label=T7 /note="T7 promoter, forward primer" promoter complement(3181..3199) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3209..3226) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3209..3225) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3218..3240) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3367..3822 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3454..3473) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3664..3685 /label=F1ori-F /note="F1 origin, forward primer" promoter 3848..3952 /gene="bla" /label=AmpR promoter CDS 3953..4813 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(4171..4190) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4984..5572 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5473..5492 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5726..5743 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5860..5881 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5896..5926 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5934..5950 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 5939..5961 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5958..5974 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5958..5974 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 5993..6013 /label=T3 /note="T3 promoter, forward primer" promoter 5995..6013 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6041..6267 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6268..6448 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6495..6620 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7113..7346 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7531..7575 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 7753..8001 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)" primer_bind 7753..7773 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 7924..7943 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 8023..8098 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 8154..8271 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct 61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat 121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa 181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg 241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt 301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg 361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc 421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt 481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt 541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat 601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc 661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca 721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc 781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg 841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc 901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac 961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt 1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag 1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag 1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat 1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt 1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg 1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc 1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca 1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg 1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca 1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc 1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc 1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg 1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg 1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc 1861 gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg 1921 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 1981 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 2041 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 2101 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 2161 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 2221 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 2281 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg 2341 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 2401 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 2461 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga 2521 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag 2581 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga 2641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 2701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 2761 ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca 2821 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc 2881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 2941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta 3001 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 3121 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc 3181 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg 3241 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc 3301 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg 3361 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 3421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 3481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 3541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 3601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 3661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 3721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 3781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg 3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 3961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 4021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 4081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 4141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 4201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 4261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 4321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 4381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 4441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 4501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 4561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 4621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 4681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 4741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 4801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 5281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 5701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 5761 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 5821 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca 5881 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag 5941 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa 6001 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc 6061 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag 6121 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa 6181 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta 6241 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg 6301 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 6361 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 6421 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag 6481 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag 6541 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga 6601 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa 6661 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca 6721 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac 6781 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat 6841 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg 6901 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg 6961 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat 7021 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga 7081 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga 7141 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta 7201 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat 7261 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa 7321 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc 7381 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat 7441 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc 7501 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat 7561 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta 7621 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata 7681 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg 7741 aggggaccca gagagggcct atttcccatg attccttcat atttgcatat acgatacaag 7801 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat 7861 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa 7921 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc 7981 ttgtggaaag gacgaaacac cgcaggcata cactgacgac aggttttaga gctagaaata 8041 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt 8101 ttttaagctt ggcgtaacta gatcttgaga caaatggcag tattcatcca caattttaaa 8161 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca 8221 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat 8281 tacagggaca gcagagatcc actttggcgc cggctcgagg ggg //

pGBW-m4252885

153832

Mammalian expression plasmid for SARS-CoV-2 S (surface glycoprotein)

SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/307333/e206d6f5-30d7-4bdf-90f4-d05f0497e65c/addgene-plasmid-153832-sequence-307333.gbk

full

LOCUS sequence-307333- 10458 bp DNA circular SYN 26-AUG-2024 DEFINITION Mammalian expression plasmid for SARS-CoV-2 S (surface glycoprotein). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10458) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 10458) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..10458 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 3824..3844 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 3845..3922 /codon_start=1 /product="calmodulin-binding peptide" /label=CBP /note="derived from skeletal muscle myosin light chain kinase; binds calmodulin with nanomolar affinity in the presence of calcium" /translation="KRRWKKNFIAVSAANRFKKISSSGAL" polyA_signal 3991..4112 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4002..4021) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4056..4075 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 4227..4273 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 4303..4439 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4484..4555 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(4781..5369) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4861..4880) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 5869..7078 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 6915..6934 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 7082..8110 /codon_start=1 /product="hygromycin B phosphotransferase" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" polyA_signal 8137..8185 /note="synthetic polyadenylation signal" CDS 8451..9311 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8669..8688) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 9324..9460 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 9612..9683 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator complement(9688..9734) /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 9752..9800 /note="synthetic polyadenylation signal" enhancer 9881..10184 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 10186..10388 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 10339..10359 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgttc gtctttcttg tgttactccc cctggtttcg tcccagtgcg tgaacttgac 61 tacgcgaacc caactgccgc cagcatatac aaatagtttc acccgtggag tatactaccc 121 tgataaagtg tttagatcaa gcgtactaca ctctactcag gacctgtttc tccccttctt 181 ctccaatgtc acatggtttc atgcgatcca cgttagcggc accaacggga caaagcgctt 241 tgacaaccct gtgctgccat tcaatgatgg tgtgtatttt gcttctaccg agaagagcaa 301 cataattcgg ggctggatct tcggaactac cctcgacagt aaaactcagt cattgctgat 361 tgtgaataac gccacaaatg tcgttatcaa ggtctgtgaa tttcaattct gcaacgatcc 421 attccttggg gtgtattacc ataaaaataa taagtcctgg atggagagcg aatttagggt 481 atactcgtct gccaacaatt gtactttcga gtatgtgtct cagcccttct taatggatct 541 ggaaggtaaa cagggcaact ttaagaacct ccgcgagttt gtattcaaaa atatagacgg 601 atactttaag atttattcca agcacacacc aatcaatttg gtgcgggatc tcccgcaagg 661 gttcagcgct ctggaacctc tagtggacct ccccatcggc attaacatca caagattcca 721 gacgctgtta gccctgcata gatcatacct tactcccggt gacagtagtt ctgggtggac 781 cgcaggcgcg gctgcatatt acgtgggata tttgcagcct cggacgtttc tgctgaaata 841 caacgagaac ggcaccatta cagatgccgt ggattgcgcc cttgaccccc taagcgaaac 901 taagtgtacg cttaaaagct tcaccgtcga gaagggaatt taccagactt ccaactttag 961 ggtccagccg acagaatcaa tagttaggtt cccaaatatc accaatttgt gcccttttgg 1021 ggaagtcttc aatgctacac gatttgcttc ggtttatgcc tggaacagga agcgtatctc 1081 aaattgcgtc gcggactaca gcgtgctcta taatagtgca agtttctcga cctttaaatg 1141 ttatggcgtg tccccgacca aactaaatga cctctgtttt acaaacgttt acgccgattc 1201 ttttgtgatt cgcggtgatg aggtacgcca aatcgcaccg ggccagactg gcaagattgc 1261 cgattacaac tacaagttgc ctgacgactt caccggatgt gttatagcct ggaatagcaa 1321 taatctggat tctaaggtcg gaggtaacta taactacctg tatagactgt tccgtaagtc 1381 aaacctgaaa ccctttgagc gggacatctc cacagagatt taccaagcag ggtctacccc 1441 ctgcaatggg gttgaaggat tcaattgcta ctttccatta cagagctacg ggttccagcc 1501 tacgaacggg gttggttacc agccttatag ggtagtggtg ctgtcgttcg aactgttgca 1561 cgctccagct acagtgtgcg ggcctaaaaa atcaactaac ctggttaaaa ataagtgtgt 1621 taattttaac ttcaatggct taaccggaac tggagtgctc acggagtcaa acaagaaatt 1681 tctaccgttt cagcaattcg ggagggatat cgcagacact acggacgcgg tcagagatcc 1741 acagaccctt gagatccttg acataacacc ctgctctttc ggaggtgtct ccgtaattac 1801 accaggcact aatacctcta atcaggtggc cgtgctttat caggatgtta actgtacaga 1861 ggtcccggtg gctatccatg ccgaccaatt gacacctacg tggcgcgtct attctaccgg 1921 ctccaacgtc ttccagaccc gggctggatg cttgataggc gcagaacacg taaacaactc 1981 ttacgaatgt gatattccca ttggagccgg gatctgcgca agctaccaga cccaaacaaa 2041 cagtcccaga cgggctcgta gcgtggcctc tcagagcata atagcctata ccatgtccct 2101 tggcgcggag aatagtgtgg catactcaaa taatagcatc gctatcccca ctaactttac 2161 tatttccgtg acgactgaga ttctgccagt gtctatgact aagacatccg tggattgcac 2221 aatgtatatt tgtggggatt ccacagagtg tagcaacttg ttgctgcaat atggatcatt 2281 ctgtacgcag ctgaatcgcg ccctcaccgg catcgctgtg gaacaggaca agaacactca 2341 agaggtcttt gctcaggtga agcaaatcta caagacccca cctataaaag acttcggggg 2401 ttttaacttc tcgcagattc tccccgaccc cagcaagcct tcaaaaagga gctttattga 2461 ggacctgttg ttcaacaagg ttactttagc cgatgccggt tttatcaaac agtacggcga 2521 ctgtcttggc gatatagcag cccgggatct gatttgcgca caaaaattca atggccttac 2581 cgtactgcca ccccttctca ccgacgaaat gatcgcgcag tacacctccg cactgctggc 2641 tgggaccatc acttcgggat ggacctttgg ggctggtgct gctttgcaaa ttcccttcgc 2701 catgcagatg gcctatcgat tcaatggcat aggggtaaca cagaacgtat tgtatgaaaa 2761 tcaaaagcta atcgctaacc agtttaattc agccattggt aagatccagg attcactctc 2821 cagtacagcc tctgccctgg gaaagctcca ggacgtggtc aaccagaacg cgcaggcatt 2881 gaatacactc gtcaaacagc tgagtagcaa tttcggcgca atcagtagtg tcttaaacga 2941 tatactgagt aggcttgata aagtagaggc cgaggtccag attgacagac taatcacggg 3001 gcgtcttcag agtttacaga cctatgtgac ccaacagctg atccgagcag cggaaattcg 3061 agcaagcgcg aacctggccg ctactaaaat gagtgagtgc gtgctgggcc aaagtaaacg 3121 agtcgacttt tgcggcaaag ggtatcatct catgtctttc cctcagtccg cccctcacgg 3181 tgttgttttc ctccacgtga catacgttcc cgcgcaggaa aagaatttta ctacggcacc 3241 cgctatttgc catgacggaa aggctcattt tccacgggag ggagtgttcg tttcaaacgg 3301 gacacactgg tttgtaactc agcggaactt ttatgagcct caaatcatca cgacagacaa 3361 taccttcgtg agtggcaact gcgatgtagt cataggcatt gtgaataaca ctgtttatga 3421 tccacttcag ccggaactcg acagcttcaa agaggaattg gataagtact ttaaaaatca 3481 taccagtcca gacgtagatc tcggcgatat atcgggcatt aacgccagtg tggtcaatat 3541 acaaaaggag attgacagac tgaatgaagt agctaagaac ctcaacgaat ctctgattga 3601 cctgcaagag ctagggaagt acgagcagta tatcaaatgg ccatggtaca tatggctggg 3661 ttttatcgcg ggtcttattg ccatagtcat ggtgacgatt atgttgtgct gtatgacctc 3721 ctgctgttct tgtttgaaag gctgctgctc ctgtggtagc tgttgcaaat tcgatgaaga 3781 tgactctgaa cccgtgttga aaggcgttaa gctacactac acagaaaacc tttactttca 3841 gtcgaagagg cgatggaaaa agaattttat cgcggtgagt gcagccaaca gattcaaaaa 3901 gatatcctcg tctggtgccc tctaaggatc cgcaggcctc tgctagcttg actgactgag 3961 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca 4021 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat 4081 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt 4141 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg 4201 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt 4261 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa 4321 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata 4381 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag 4441 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct 4501 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc 4561 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat 4621 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt 4681 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc 4741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 4801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 4861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 4921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 4981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 5161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 5221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 5281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 5341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5581 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg 5641 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc 5701 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt 5761 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca 5821 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc 5881 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg 5941 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc 6001 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg 6061 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct 6121 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga 6181 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt 6241 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc 6301 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt 6361 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg 6421 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca 6481 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga 6541 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt 6601 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt 6661 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt 6721 tctgttggct tataatgcag ggtggggcca ccagccggta ggtgtgcggt aggcttttct 6781 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga 6841 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta 6901 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt 6961 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca 7021 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa 7081 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt 7141 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt 7201 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa 7261 agatcgttat gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga 7321 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac 7381 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat 7441 ggatgccatc gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca 7501 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt 7561 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga 7621 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt 7681 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga 7741 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt 7801 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc 7861 gccccggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt 7921 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc 7981 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga 8041 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc 8101 aaaggaataa gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga 8161 tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga 8221 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc 8281 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc 8341 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc 8401 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc 8461 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 8521 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt 8581 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct 8641 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 8701 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt 8761 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 8821 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga 8881 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 8941 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 9001 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac 9061 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 9121 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 9181 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 9241 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 9301 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc 9361 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc 9421 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact 9481 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca 9541 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc 9601 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat 9661 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc 9721 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt 9781 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa 9841 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa 9901 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 9961 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 10021 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 10081 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 10141 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc 10201 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 10261 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 10321 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 10381 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac 10441 acacccgcca gcggccgc //

pXD70Tet(DadRmt7)-N-3xFLAG-DadR(linker)

191643

E. coli - Eggerthella lenta shuttle plasmid (TetR), N-3xFLAG-dadR(linker region shuffled) complementation

Emily Balskus

Dong et al Nat Commun. 2022 Dec 9;13(1):7624. doi: 10.1038/s41467-022-33576-3.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/375874/5e219fbe-5a3e-43fd-a087-1737d41fb31b/addgene-plasmid-191643-sequence-375874.gbk

full

LOCUS sequence-375874- 9110 bp DNA circular SYN 27-AUG-2024 DEFINITION E. coli - Eggerthella lenta shuttle plasmid (TetR), N-3xFLAG-dadR(linker region shuffled) complementation. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9110) AUTHORS Dong X, Guthrie BGH, Alexander M, Noecker C, Ramirez L, Glasser NR, Turnbaugh PJ, Balskus EP TITLE Genetic manipulation of the human gut bacterium Eggerthella lenta reveals a widespread family of transcriptional regulators. JOURNAL Nat Commun. 2022 Dec 9;13(1):7624. doi: 10.1038/s41467-022-33576-3. PUBMED 36494336 REFERENCE 2 (bases 1 to 9110) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun."; date: "2022-12-9"; volume: "13(1):7624. doi"; pages: " 10.1038/s41467-022-33576-3" FEATURES Location/Qualifiers source 1..9110 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(4972..4999) /label=T7Te terminator /note="phage T7 early transcription terminator" CDS complement(6508..6573) /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" misc_feature 6817..6836 /label=BioBrick prefix /note="BioBrick prefix for parts that start with ""ATG""" primer_bind complement(6993..7011) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7079..7183 /gene="bla" /label=AmpR promoter CDS 7184..8044 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(7402..7421) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8215..8803 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8704..8723 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator complement(8984..9055) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ttatagcatg gcggcaaacg ctggcgagac gaaggtgaag ctctgggagg gcatctgcta 61 cccagagaac atgcgcgatg actggcagga cgagatagac gacattcttc aagtgccttt 121 cgcttatgcg gttcacgaca tagaccatga ccagaagagc aagcagcgca agacgcatgc 181 gcacgtcatt gttgtctggg gcggcaacac gacacgcaag gcaatcatca acgtcttgaa 241 ccgattgagc gcggacggca agaagtgctg ttcgtcagcc gagccaatca acaacatcag 301 gcatgcgtat gactatctga ttcacgacac ggcttcctgt cgcaagaagg gcaaggagct 361 gtatccggtc gaagcacgca tcgagggaaa caacttcgac attggtcagc tcgaacagct 421 ctcgactaag gacaagcagg acatgctctt cgagttggtc ggcttcatca tgtgcgagag 481 gttcgagacc atcaacgatt tcacggctgc ggcgctgagg gagtttcccg agcagtaccg 541 cgaaatcatc gtggggtaca actcgattct cgagcgctat tgtcgcggga actatctgaa 601 cgccgaacgt aagcgcaagt gcgtggaaca gtcaggtgcc aaagagtagt cgggtcaatg 661 cagcaatgcg tgactacggg gcggcgtggt ctgcgctcgc gcagatctaa gccgactcgt 721 tcacggattg cggaattgac gcgacggcca accgaataga ccagacttga ttaagggtcg 781 atggacgctc tgctccatcg accctttcct tgcccgtgtc tcgactggat gagacctgca 841 gaatcgggcg gctcattatt tttcgctaac tacgtccgaa aaataatgtg tccgaaaaat 901 aatgtgacaa atgacaaatg acaaatgacc acaggggggc cactacgacg gcccccctaa 961 tagtgccgtg tgccgtgtgc cgaaccgcag gtagattgag gggtggttcg gaggggcgaa 1021 gtccctccgt tcggcgctcg cgccgatggc ccgagggccg tgagcgaagc cgcgaacggg 1081 caagggctgg ggggtctcgg ggggaacgcc cgagcgggtg cagggctggc cctgccacgt 1141 ccttagcgta gctaaggtct aacgtgctag actttagtcg attaagtctg tacggttcgg 1201 aggtttccat ggctcacatc gcgaaataca aggctccatc ggtcgggcac atgcttgccc 1261 actaccgccg cgaccgcagc agcctggagc gcgacaacat cgaccccgag cgcatcaaga 1321 acgacttggt ggtcggccac tacacgaaca aggacggcaa gctcgttgtg gggcgcgtgg 1381 agccgcgcga tggcgagccg aactggggca cggtggagag ccgcatcgag cgcgtgaacg 1441 aggcccagaa ggccgcaggg aagcgtgcca cgcgcaagga tgcggtcgtt atggctgacg 1501 ttgttgtgac gcttcccgac aatgtgcgca agggcgatga ggacaggttc ttccgtctca 1561 cgtactggta cctgtcgaag aagttcggca tcgagaacat gatgggcggg tacgtccaca 1621 aggacgaggt gttgaaggat ggcacgcccg cccgcgacca catgcacgtg ccgttcacgc 1681 ccatcctaga tggacggttc aactacaagc agatgtgccc ccgcagcttc taccggagca 1741 tgcacaagga gcttggggat tacctagagg ggcgcatggg ctaccgccct gcaatcgagc 1801 ttgacgagga gacgcgggcg cagcgtgtat acacggacaa gtccgtggat atcgacaagg 1861 tacgctgtgc ggtcgataag gccgtggttg aacctgcgaa ggacgaagcg gcgcgaatcg 1921 tcgcggcggc tagggaagaa gccgccgctt tactgaatga tgccgaaacg cgtaaggcag 1981 agcttgttac cgagattgcc gacaaggagg gtgaccttgc cgagcttgac agccagctcg 2041 aggacgtgaa gctcgatatc gaagatgagc aagaccgatt ggagtgtctt cggcaacgag 2101 cggacggagt tgcgcgagac attggagagc ttcaacccat cgctaccgag gttcggggct 2161 gggaagctgc gggaaaagct gagcgcggcg caatcctcga taacatcgtt gttaagtgcg 2221 atggactcgc gagccgcatc agagcaggag ttgcgggaat gcgaatcaaa gtggaagagc 2281 ttagaagtcg aatatcgcgc ccgctcgaat atctgatgcg cagggaacag ccaaggcaac 2341 agagccttaa tgacgtgctg cgcgatgcgc agcgggcggc ggatgcgtgg aaccgcgacc 2401 acgctgcacc gcagaggacg tatcgcgggc gggctagatg atgtgttata gtagtacctg 2461 taatacacga caaggaggtg cgacgcgaaa aaaccccgct gaagcggggt tttttgcgca 2521 ccacagcaga cagcagaaaa cattctgaac gctaggaagc cggtatgatt gttacatata 2581 aggggaagaa aaatttcttt taggtacttg ctttcctaaa actgatgtga tacaatgatt 2641 taatccagaa aaggagtaaa aaatatgcgg caaggtattc ttaaataaaa ctataatcaa 2701 atagtgggaa caaaggatta tgatagctcc ttttgtaggg acttagtttt ttgtacccaa 2761 tttaagaata cttttgcctt atcaattttg acatatcccc aaaaacagca atcacaaaca 2821 ggtgtatgct gtatatgtgt atgtccgcaa cttataatcc ccagtggtaa aagtatttta 2881 ctgctgggga tttttatgcc ctttggggct gtaaagggag gacaatcaca tgaaaataat 2941 caatattgga attcttgccc atgtagacgc tggaaagacg accttgacgg agagcctgct 3001 atatgccagc ggagccattt cagaaccggg gagcgtcgaa aaagggacaa cgaggacgga 3061 caccatgttt ttggagcggc agcgtgggat taccattcaa gcggcagtca cttccttcca 3121 gtggcacaga tgtaaagtta acattgtgga tacgcccggc cacatggatt ttttggcgga 3181 ggtgtaccgc tctttggctg ttttagatgg ggccatcttg gtgatctccg ctaaagatgg 3241 cgtgcaggcc cagacccgta ttctgttcca tgccctgcgg aaaatgaaca ttcccaccgt 3301 tatctttatc aacaagatcg accaggctgg cgttgatttg cagagcgtgg ttcagtctgt 3361 tcgggataag ctctccgccg atattatcat caagcagacg gtgtcgctgt ccccggaaat 3421 agtcctggag gaaaataccg acatagaagc atgggatgcg gtcatcgaaa ataacgatga 3481 attattggaa aagtatatcg caggagaacc aatcagccgg gaaaaacttg cgcgggagga 3541 acagcagcgg gttcaagacg cctccctgtt cccagtctat catggcagcg ccaaaaatgg 3601 ccttggcatt caaccgttga tggatgcggt gacagggctg ttccaaccga ttggggaaca 3661 ggggggcgcc gccctatgcg gcagcgtttt caaggttgag tacaccgatt gcggccagcg 3721 gcgtgtctat ctacggttat acagcggaac gctgcgcctg cgggatacgg tggccctggc 3781 cgggagagaa aagctgaaaa tcacagagat gcgtattcca tccaaagggg aaattgttcg 3841 gacagacacc gcttatcagg gtgaaattgt tatccttccc agcgacagcg tgaggttaaa 3901 cgatgtatta ggggaccaaa cccggctccc tcgtaaaagg tggcgcgagg accccctccc 3961 catgctgcgg acggcgattg cgccgaaaac ggcagcgcaa agagaacggc tgctggacgc 4021 tcttacgcaa cttgcggata ctgacccgct tttgcgttgc gaagtggatt ccatcaccca 4081 tgagatcatt ctttcttttt tgggccgggt gcagttggag gttgtttccg ctttgctgtc 4141 ggaaaaatac aagcttgaaa cagtggtaaa ggaaccctcc gtcatttata tggagcggcc 4201 gctcaaagca gccagccaca ccatccatat cgaggtgccg cccaacccgt tttgggcatc 4261 cataggactg tctgttacac cactctcgct tggctccggt gtacaatacg agagccgggt 4321 ttcgctggga tacttgaacc agagttttca aaacgctgtc agggatggta tccgttacgg 4381 gctggagcag ggcttgttcg gctggaacgt aacggactgt aagatttgct ttgaatacgg 4441 gctttattac agtccggtca gcacgccggc ggacttccgc tcattggccc cgattgtatt 4501 ggaacaggca ttgaaggaat cggggacgca gctgctggaa ccttatctct ccttcatcct 4561 ctatgcgccc caggaatacc tttccagggc ttatcatgat gcaccgaaat actgtgccac 4621 catcgaaacg gcccaggtaa aaaaggatga agttgtcttt actggcgaga ttcccgcccg 4681 ctgtatacag gcataccgta ctgatctggc cttttacacc aacgggcgga gcgtatgcct 4741 tacagagctg aaaggatatc aggccgctgt cggtcagccg gtcatccagc cccgccgtcc 4801 aaacagccgc ctggacaagg tgcgccatat gtttcagaag gtaatgtaaa tcgttaaggg 4861 atcaactttg ggagagagtt caaaattgat ccttttttta taacaggaat taaataataa 4921 aaaagccgga ttaataatct ggctttttat attctctctc tagtatataa acgcagaaag 4981 gcccacccga aggtgagcca gtgtgagtcg actagttcag accgcctcgt cctgcgcggc 5041 cgtctcgtgg gccaggtcga tgagctcctg gcgattgtcc acgcccgtct tctggtagat 5101 gtggcggata tgcgtgctca ccgtgcccgc cgagatgaac agcgtctcct ggatgcgcga 5161 gatggtgcgg ccctccagca gcagcggaag cacgtcggtc tcgcgcggcg acagggcgaa 5221 gcgctccgcg atcagcgcgc acggggcgtc cgggtcgtcg cccgtggggg gttcggcggc 5281 cacctccaca tccatcaggc tgacctcgcc gatgccgatt ttgatgaggt cggtctccgt 5341 gaacaggaac aaatgcgcga tgaacagcag cgacacggcc accagcgtga ccagcgtgag 5401 gtcgaacatg ctgaccccgg tcgcctccag cgaatgggcg aacagctggc ctgcggtctc 5461 gccgccgtac agcgccacga accccaggcc gatcgtccgc gccgggctca cgcggaagct 5521 ggacgccacg tcgatgcaca ccacgcacag caccagcacg aagcagtggt agccgccgaa 5581 gataagcgcg ctcgaatacg tgttgttgtc ggccatgaac ggcgtggaca agcatcccaa 5641 gcacagcatc agcatggaga cgcggtacga gaagaacacg ctgtcgcgct gcaggaacac 5701 ggctatcagg cagatggcca acgctccgat gaagcccgcc agaaacaggt acgtgccgaa 5761 gatgtcctgc acctcgtagg ttttgaaccc cgacaccacg tcgaagaacc ccatcaacat 5821 gccgatgagc accgcgccca gaagcgcctt cgcgaacagg gtcgccgtat ggcgcgacaa 5881 aggctgctcg ggctcgcgcg gcagctgcgg cgtatcgggc gcgctgcgcc tgagagctgc 5941 cgccgaaagc acgggcatga caacgatgag cacgatgacg ggtatcgccg gccctacgat 6001 cagcaagaac cccacgacga acgcgatgca cagcgacgag gcaagctcca caagcgccaa 6061 gcggcctccc agccccgact cgagctcgat ccagcacaag aacagcacgg ccgagcccgt 6121 gccggtcatg accgcgctca ggcccagcac cagcatgccg acgggcgtgg taggatcgga 6181 gaacggcgtg agcacgctgc ccgcgctcat catgatggcc gatcccagca cgatggagcg 6241 gttgccgctc acccgctcgc gccgcacgat gacggagacg aacagcatgg tcaccaccat 6301 ggccaccagc gatatgatca ggctgacgta cagcgagccg cgcacgtccg acgagaagcc 6361 gtacggcacg atgctgtaga acatcatgag catccaaagc cattgcaagg ccgcgcccgc 6421 gatgcgcttg agggaaaggc tcgtccccaa cgcggtatcc catgcactcc gaagcttttc 6481 ggatccgcgg ccgctacgta gaattccctt gtcatcgtca tccttgtaat cgatgtcatg 6541 atctttataa tcaccgtcat ggtctttgta gtccatgccc caccccctgt gagcccgtac 6601 ataacgtcgc tcgcgccgac ccctctgtcg acgggttcga ttatacggaa aagatgagcg 6661 gccctctttg aggaaactat tctagtttca gaataattta tgacctacgg ttaaagtttc 6721 cccagttcaa agcactatcg aatttttcat gatcacaaga tcataagaat ttatgatgcg 6781 gtcgaacctg ctttccaacc tttggacgat gctcatctag aagcggccgc gaattcagaa 6841 tagtaataca ggatccgaat cgtttcagtt gccgccggat taataatcaa ataaccgttc 6901 ctaaaaaatg ccctcttggg ttatcaagag ggtcattata tttcgcaaaa aaccccgctt 6961 cggcggggtt ttttcgcctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt 7021 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg 7081 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 7141 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 7201 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 7261 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 7321 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 7381 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca 7441 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 7501 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 7561 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 7621 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 7681 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac 7741 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 7801 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 7861 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 7921 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 7981 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 8041 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 8101 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 8161 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 8221 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 8281 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 8341 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa 8401 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 8461 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 8521 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 8581 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 8641 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 8701 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 8761 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 8821 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag 8881 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg 8941 tgaatccgtt agcgaggtgc cgccggcatg agctcctcta gtagagagcg ttcaccgaca 9001 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgaaaaa 9061 gaaaagcccc agcaagcttg gcggcagcag gggcattcga gacaaggaga //

pDONR221_50_100_CA_repeat

65867

Carries CA repeats of 50-100 nucleotides in length, an RBPMS target

Thomas Tuschl

RNA-binding protein 3' UTR target plasmids (unpublished) (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/116930/f5abf171-1982-4e07-b2c8-e2084e6448ca/addgene-plasmid-65867-sequence-116930.gbk

partial

LOCUS sequence-116930- 779 bp DNA linear UNA 26-AUG-2024 DEFINITION Carries CA repeats of 50-100 nucleotides in length, an RBPMS target. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 779) TITLE RNA-binding protein 3' UTR target plasmids (unpublished) JOURNAL Unpublished REFERENCE 2 (bases 1 to 779) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" FEATURES Location/Qualifiers source 1..779 /mol_type="genomic DNA" /organism="unspecified" primer_bind complement(256..272) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(256..272) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(269..291) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 280..296 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(304..334) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 349..370 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(487..504) /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(738..757) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt caggccgtgt 61 gtgtgtgtgt gtgtggtgcc gtgtgtgtgt gtgtgtgtgg tgccgtgtgt gtgtgtgtgt 121 gtggtgccgt gtgtgtgtgt gtgcctgaat tccagcacac tggcggccgt tactagtgga 181 tccgagctcg gtaccaagct tgatgcatag cttgagtatt ctaacgcgtc acctaaatag 241 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 301 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 361 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 421 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 481 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 541 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 601 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 661 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 721 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgct //

pLPCX-IRES-GFP

65436

Low

For transfection or retroviral production. Mammalian expression of EGFP driven by the ECMV IRES.

Trond Aasen

Salat-Canela et al Cell Commun Signal. 2014 May 8;12:31. doi: 10.1186/1478-811X-12-31.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/110889/1875ccce-bf02-4a82-95a1-eb6ba2f8ee85/addgene-plasmid-65436-sequence-110889.gbk

full

LOCUS sequence-110889- 7579 bp DNA circular SYN 25-AUG-2024 DEFINITION For transfection or retroviral production. Mammalian expression of EGFP driven by the ECMV IRES.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7579) AUTHORS Salat-Canela C, Sese M, Peula C, Ramon y Cajal S, Aasen T TITLE Internal translation of the connexin 43 transcript. JOURNAL Cell Commun Signal. 2014 May 8;12:31. doi: 10.1186/1478-811X-12-31. PUBMED 24884945 REFERENCE 2 (bases 1 to 7579) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Commun Signal."; date: "2014-05-8"; volume: "12:31. doi"; pages: " 10.1186/1478-811X-12-31" FEATURES Location/Qualifiers source 1..7579 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 414..589 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 652..851 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1052..1468 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1398..1420 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1436..1452 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1566..2165 /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" primer_bind complement(1566..1585) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2062..2082 /label=Puro-F /note="Puromycin resistance gene, forward primer" enhancer 2340..2643 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2644..2847 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2798..2818 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 2888..2944 /label=MCS /note="multiple cloning site" misc_feature 2946..3532 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3113..3130) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3340..3359 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3533..4252 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3533..4249 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3578..3599) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3839..3858) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4186..4207 /label=EGFP-C /note="EGFP, forward primer" LTR 4318..4911 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(5014..5036) /label=pGEX 3' /note="pGEX vectors, reverse primer" protein_bind 5164..5197 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 5257..5290 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." primer_bind 5310..5329 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5410..5427) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5581..6169) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5661..5680) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6340..7200) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6963..6982 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7201..7305) /gene="bla" /label=AmpR promoter primer_bind 7373..7391 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat 61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc 121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca 181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg 241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa 301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac 361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa 421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac 481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg 541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt 601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc 661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg 721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt 781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg 841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt 901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa 961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct 1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt 1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa 1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc 1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc 1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt 1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc 1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta 1441 tccagccctc actccttctc taggcgccgg aattgatccc ccgggctgca ggtcggccgc 1501 cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca aggagacgac 1561 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg 1621 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc 1681 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc 1741 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc 1801 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga 1861 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca 1921 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc 1981 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct 2041 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca 2101 ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg 2161 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct 2221 ccgaccgaag ccgacccggg cggccccgcc gaccccgcac ccgcccccga ggcccaccga 2281 ctctagtaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 2341 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 2401 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 2461 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 2521 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 2581 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 2641 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 2701 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 2761 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 2821 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac 2881 cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg 2941 atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc 3001 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg 3061 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc 3121 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga 3181 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg 3241 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag 3301 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc 3361 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct 3421 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac 3481 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag 3541 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 3601 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 3661 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 3721 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 3781 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 3841 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 3901 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 3961 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 4021 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 4081 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 4141 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 4201 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg 4261 cctcggccaa acatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat 4321 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg 4381 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc 4441 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag 4501 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc 4561 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga 4621 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga 4681 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata 4741 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt 4801 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt 4861 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc 4921 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg 4981 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 5041 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 5101 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta 5161 ctgataactt cgtataatgt atgctatacg aagttattag gtctgaagag gagtttacgt 5221 ccagccaagc tagggccgcg atccggaacc cttaatataa cttcgtataa tgtatgctat 5281 acgaagttat cagtactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 5341 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc 5401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 5881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 5941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6001 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg 6661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 6781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 6841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 6901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg 6961 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7021 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7081 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7141 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7201 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7261 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7321 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 7381 tatcacgagg ccctttcgtc ttcaagaatt gctagcaatt gctagcaatt gctagcaatt 7441 cataccagat caccgaaaac tgtcctccaa atgtgtcccc ctcacactcc caaattcgcg 7501 ggcttctgcc tcttagacca ctctacccta ttccccacac tcaccggagc caaagccgcg 7561 gcccttccgt ttctttgct //

DH-BB(5'SIN;TE12ORF)

72309

Low

Defective Helper to produce neurotropic, propagation-incompetent Sindbis virus

Anthony Zador

DH-BB(5'SIN;TE12ORF) (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/145921/8dd88aa2-a46a-4f54-9665-602d9c550e32/addgene-plasmid-72309-sequence-145921.gbk

full

LOCUS sequence-145921- 6729 bp DNA circular SYN 25-AUG-2024 DEFINITION Defective Helper to produce neurotropic, propagation-incompetent Sindbis virus. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6729) TITLE DH-BB(5'SIN;TE12ORF) JOURNAL Unpublished REFERENCE 2 (bases 1 to 6729) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" FEATURES Location/Qualifiers source 1..6729 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 4900..5004 /gene="bla" /label=AmpR promoter CDS 5005..5865 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5223..5242) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6036..6624 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6525..6544 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 attgacggcg tagtacacac tattgaatca aacagccgac caattgcact accatcacaa 61 tggagaagcc agtagtaaac gtagacgtag acccccagag tccgtttgtc gtgcaactgc 121 aaaaaagctt cccgcaattt gaggtagtag cacagcaggt cactccaaat gaccatgcta 181 atgccagagc attttcgcat ctggccagta aactaatcga gctggaggtt cctaccacag 241 cgacgatctt ggacataggc agcgcaccgg ctcgtagaat gttttccgag caccagtatc 301 attgtgtctg ccccatgcgt agtccagaag acccggaccg catgatgaaa tatgccagta 361 aactggcgga aaaagcgtgc aagattacaa acaagaactt gcatgagaag attaaggatc 421 tccgggatcc cctgaaaagg ctgtttaagt tgggtaaacc gctcccagcc gacgacgagc 481 aagacgaaga cagaagacgc gctctgctag atgaaacaaa ggcgtggttt agagtaggta 541 taacaggcac tttagcagtg gccgtgacga cccggtatga ggtagacaat attacacctg 601 tcctactggc attgagaact tttgcccaga gcaaaagagc attccaagcc atcagagggg 661 aaataaagca tctctacggt ggtcctaaat agtcagcata gtacatttca tctgactaat 721 actacaacac caccaccatg aatagaggat tctttaacat gctcggccgc cgccccttcc 781 cggcccccac tgccatgtgg aggccgcgga gaaggaggca ggcggccccg atgcctgccc 841 gcaacgggct ggcttctcaa atccagcaac tgaccacagc cgtcagtgcc ctagtcattg 901 gacaggcaac tagacctcaa cccccacgtc cacgcccgcc accgcgccag aagaagcagg 961 cgcccaagca accaccgaag ccgaagaaac caaaaacgca ggagaagaag aagaagcaac 1021 ctgcaaaacc caaacccgga aagagacagc gcatggcact taagttggag gccgacagat 1081 tgttcgacgt caagaacgaa gacggagatg tcatcgggca cgcactggcc atggaaggaa 1141 aggtaatgaa acctctgcac gtgaaaggaa ccatcgacca ccctgtgcta tcaaagctca 1201 aatttaccaa gtcgtcagca tacgacatgg agttcgcaca gttgccagtc aacatgagaa 1261 gtgaggcatt cacctacacc agtgaacacc ccgaaggatt ctataactgg caccacggag 1321 cggtgcagta tagtggaggt agatttacca tccctcgcgg agtaggaggc agaggagaca 1381 gcggtcgtcc gatcatggat aactccggtc gggttgtcgc gatagtcctc ggtggcgctg 1441 atgaaggaac acgaactgcc ctttcggtcg tcacctggaa tagtaaaggg aagacaatta 1501 agacgacccc ggaagggaca gaagagtggt ccgcagcacc actggtcacg gcaatgtgtt 1561 tgctcggaaa tgtgagcttc ccatgcgacc gcccgcccac atgctatacc cgcgaacctt 1621 ccagagccct cgacatcctt gaagagaacg tgaaccatga ggcctacgat accctgctca 1681 atgccatatt gcggtgcgga tcgtctggca gaagcaaaag aagcgtcact gacgacttta 1741 ccctgaccag cccctacttg ggcacatgct cgtactgcca ccatactgaa ccgtgcttca 1801 gccctgttaa gatcgagcag gtctgggacg aagcggacga taacaccata cgcatacaga 1861 cttccgccca gtttggatac gaccatagcg gagcagcaag cgcaaacaag taccgctaca 1921 tgtcgcttaa gcaggatcac accgttaaag aaggcaccat ggatgacatc aagattagca 1981 cctcaggacc gtgtagaagg cttagctaca aaggatactt tctcctcgca aaatgccctc 2041 caggggacag cgtaacggtt agcatagtga gtagcaactc agcaacgtca tgtacactgg 2101 cccgcaagat aaaaccaaaa ttcgtgggac gggaaaaata tgatctacct cccgttcacg 2161 gtaaaaaaat tccttgcaca gtgtacgacc gtctgaaaga aacaactgca ggctacatca 2221 ctatgcacag gccgggaccg cacgcttata catcctacct ggaagaatca tcagggaaag 2281 tttacgcaaa gccgccatct gggaagaaca ttacgtatga gtgcaagtgc ggcgactaca 2341 agaccggaac cgtttcgacc cgcaccgaaa tcactggttg caccgccatc aagcagtgcg 2401 tcgcctataa gagcgaccaa acgaagtggg tcttcaactc accggacttg atcagacatg 2461 acgaccacac ggcccaaggg aaattgcatt tgcctttcaa gttgatcccg agtacctgca 2521 tggtccctgt tgcccacgcg ccgaatgtaa tacatggctt taaacacatc agcctccaat 2581 tagatacaga ccacttgaca ttgctcacca ccaggagact aggggcaaac ccggaaccaa 2641 ccactgaatg gatcgtcgga aagacggtca gaaacttcac cgtcgaccga gatggcctgg 2701 aatacatatg gggaaatcat gagccagtga gggtctatgc ccaagagtca gcaccaggag 2761 accctcacgg atggccacac gaaatagtac agcattacta ccatcgccat cctgtgtaca 2821 ccatcttagc cgtcgcatca gctaccgtgg cgatgatgat tggcgtaact gttgcagtgt 2881 tatgtgcctg taaagcgcgc cgtgagtgcc tgacgccata cgccctggcc ccaaacgccg 2941 taatcccaac ttcgctggca ctcttgtgct gcgttaggtc ggccaatgct gaaacgttca 3001 ccgagaccat gagttacttg tggtcgaaca gtcagccgtt cttctgggtc cagttgtgca 3061 tacctttggc cgctttcatc gttctaatgc gctgctgctc ctgctgcctg ccttttttag 3121 tggttgccgg cgcctacctg gcgaaggtag acgcctacga acatgcgacc actgttccaa 3181 atgtgccaca gataccgtat aaggcacttg ttgaaagggc agggtatgcc ccgctcaatt 3241 tggagatcac tgtcatgtcc tcggaggttt tgccttccac caaccaagag tacattacct 3301 gcaaattcac cactgtggtc ccctccccaa aaatcaaatg ctgcggctcc ttggaatgtc 3361 agccggccgc tcatgcagac tatacctgca aggtcttcgg aggggtctac ccctttatgt 3421 ggggaggagc gcaatgtttt tgcgacagtg agaacagcca gatgagtgag gcgtacgtcg 3481 aattgtcagc agattgcgcg tctgaccacg cgcaggcgat taaggtgcac actgccgcga 3541 tgaaagtagg actgcgtata gtgtacggga acactaccag tttcctagat gtgtacgtga 3601 acggagtcac accaggaacg tctaaagact tgaaagtcat agctggacca atttcagcat 3661 catttacgcc attcgatcat aaggtcgtta tccatcgcgg cctggtgtac aactatgact 3721 tcccggaata tggagcgatg aaaccaggag cgtttggaga cattcaagct acctccttga 3781 ctagcaagga tctcatcgcc agcacagaca ttaggctact caagccttcc gccaagaatg 3841 tgcatgtccc gtacacgcag gccgcatcag gatttgagat gtggaaaaac aactcaggcc 3901 gcccattgca ggaaaccgca cctttcgggt gtaagattgc agtaaatccg ctccgagcgg 3961 tggactgttc atacgggaac attcccattt ctattgacat cccgaacgct gcctttatca 4021 ggacatcaga tgcaccactg gtctcaacag tcaaatgtga agtcagtgag tgcacttatt 4081 cagcagactt cgacgggatg gccaccctgc agtatgtatc cgaccgcgaa ggtcaatgcc 4141 ccgtacattc gcattcgagc acagcaactc tccaagagtc gacagtacat gtcctggaga 4201 aaggagcggt gacagtacac tttagcaccg cgagtccaca ggcgaacttt atcgtatcgc 4261 tgtgtgggaa gaagacaaca tgcaatgcag aatgtaaacc accagctgac catatcgtga 4321 gcaccccgca caaaaatgac caagaatttc aagccgccat ctcaaaaaca tcatggagtt 4381 ggctgtttgc ccttttcggc ggcgcctcgt cgctattaat tataggactt atgatttttg 4441 cttgcagcat gatgctgact agcacacgaa gatgaccgct acgccccaat gatccgacca 4501 gcaaaactcg atgtacttcc gaggaactga tgtgcataat gcatcaggct ggtacattag 4561 atccccgctt accgcgggca atatagcaac actaaaaact cgatgtactt ccgaggaagc 4621 gcagtgcata atgctgcgca gtgttgccac ataaccacta tattaaccat ttatctagcg 4681 gacgccaaaa actcaatgta tttctgagga agcgtggtgc ataatgccac gcagcgtctg 4741 cataactttt attatttctt ttattaatca acaaaatttt gtttttaaca tttcaaaaaa 4801 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa agggaattcc tcgaggggaa ttaattcttg 4861 aagacgaaag ggccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 4921 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 4981 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 5041 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 5101 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 5161 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 5221 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc 5281 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 5341 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 5401 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 5461 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 5521 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 5581 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 5641 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 5701 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 5761 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 5821 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 5881 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 5941 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 6001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 6061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 6121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 6181 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 6241 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 6301 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 6361 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 6421 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 6481 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 6541 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 6601 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcga gctcgtatgg acatattgtc 6661 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttagggg 6721 acactatag //

lenti-SpCas9 puro

104994

Low

This 3rd generation lentiviral construct delivers hSpCas9 and puromycin resistance.

Brett Stringer

Stringer et al Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/212630/167aa445-4112-47bf-a347-25352c796ce0/addgene-plasmid-104994-sequence-212630.gbk

full

LOCUS sequence-212630- 12671 bp DNA circular SYN 25-AUG-2024 DEFINITION This 3rd generation lentiviral construct delivers hSpCas9 and puromycin resistance.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12671) AUTHORS Stringer BW, Day BW, D'Souza RCJ, Jamieson PR, Ensbey KS, Bruce ZC, Lim YC, Goasdoue K, Offenhauser C, Akgul S, Allan S, Robertson T, Lucas P, Tollesson G, Campbell S, Winter C, Do H, Dobrovic A, Inglis PL, Jeffree RL, Johns TG, Boyd AW TITLE A reference collection of patient-derived cell line and xenograft models of proneural, classical and mesenchymal glioblastoma. JOURNAL Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z. PUBMED 30894629 REFERENCE 2 (bases 1 to 12671) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; date: "2019-03-20"; volume: "9(1):4902. doi"; pages: " 10.1038/s41598-019-41277-z" FEATURES Location/Qualifiers source 1..12671 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 242..621 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 623..821 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 776..796 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 839..1019 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1066..1191 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1684..1917 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2102..2146 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2444..2561 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2673..2884 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 2903..2912 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 2909..7012 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 2909..7012 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 7013..7060 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 7061..7084 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 7094..7150 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 7151..7747 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind 7644..7664 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 7763..8351 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7816..7836) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8234..8245) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 8423..8656 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(8682..8699) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8688..8912 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 8958..9386 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9045..9064) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9255..9276 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(9395..9415) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9400..9729 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9580..9715 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9642..9661 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 9777..9824 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 9843..10217 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 10347..10468 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10384..10403) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10438..10457 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10517..10533) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10517..10533) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10530..10552) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10541..10557 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(10565..10595) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10610..10631 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(10748..10765) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10919..11507) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10999..11018) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11678..12538) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 12301..12320 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12539..12643) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgcgcgt 241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc 301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg 841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 1021 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg 1081 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca 1141 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag 1201 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa 1261 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct 1321 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt 1381 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg 1441 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa 1501 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga 1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc 2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga 2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact 2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg 2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa 2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca 2581 gcagagatcc agtttggtta attaaggtac cctcgagtgt acagctagct aggtcttgaa 2641 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc 2701 cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt 2761 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc 2821 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac 2881 acaggaccgg ttctagagcg ctgccaccat ggacaagaag tacagcatcg gcctggacat 2941 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa 3001 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct 3061 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag 3121 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc 3181 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa 3241 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa 3301 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct 3361 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga 3421 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac 3481 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat 3541 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg 3601 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa 3661 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta 3721 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct 3781 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga 3841 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga 3901 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt 3961 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga 4021 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt 4081 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc 4141 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc 4201 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta 4261 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga 4321 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt 4381 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca 4441 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac 4501 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct 4561 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa 4621 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct 4681 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga 4741 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat 4801 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct 4861 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg 4921 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag 4981 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc 5041 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc 5101 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat 5161 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca 5221 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct 5281 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct 5341 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa 5401 ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc 5461 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc 5521 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct 5581 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact 5641 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt 5701 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg 5761 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca 5821 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc 5881 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg 5941 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa 6001 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac 6061 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga 6121 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca 6181 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct 6241 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta 6301 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa 6361 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag 6421 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa 6481 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa 6541 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa 6601 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga 6661 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga 6721 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct 6781 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca 6841 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat 6901 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag 6961 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaagcgacc 7021 tgccgccaca aagaaggctg gacaggctaa gaagaagaaa gattacaaag acgatgacga 7081 taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa 7141 tcctggaccg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag 7201 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga 7261 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg 7321 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac 7381 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt 7441 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc 7501 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg 7561 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc 7621 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt 7681 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg 7741 tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac 7801 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 7861 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 7921 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 7981 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 8041 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 8101 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 8161 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 8221 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 8281 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 8341 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac 8401 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg 8461 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc 8521 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg 8581 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg 8641 tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag 8701 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 8761 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 8821 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 8881 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 8941 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 9001 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 9061 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 9121 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 9181 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 9241 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 9301 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 9361 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 9421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 9481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 9541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 9601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 9661 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 9721 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg 9781 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa 9841 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg 9901 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg 9961 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg 10021 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg 10081 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc 10141 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg 10201 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa 10261 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 10321 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 10381 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 10441 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 10501 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 10561 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 10621 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 10681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 10741 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 10801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 10861 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 10921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 10981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 11041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 11101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 11161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 11221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 11281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 11341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 11401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 11461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 11521 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 11581 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 11641 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 11701 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 11761 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 11821 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 11881 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 11941 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 12001 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 12061 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 12121 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 12181 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 12241 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 12301 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 12361 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 12421 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 12481 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 12541 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 12601 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 12661 tgccacctga c //

pHM019Env

137685

Express HIV Env which is isolated a from IRB approved HIV infected patient samples from Singapore and cloned into expression vector (pcDNA 3.1)

Anjali Joshi

Joshi et al Virology. 2017 Dec;512:222-233. doi: 10.1016/j.virol.2017.09.026. Epub 2017 Oct 9.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/297219/cacaf556-5345-4c40-a0f4-ea971ee405d6/addgene-plasmid-137685-sequence-297219.gbk

full

LOCUS sequence-297219- 8471 bp DNA circular SYN 26-AUG-2024 DEFINITION Express HIV Env which is isolated a from IRB approved HIV infected patient samples from Singapore and cloned into expression vector (pcDNA 3.1). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8471) AUTHORS Joshi A, Cox EK, Sedano MJ, Punke EB, Lee RT, Maurer-Stroh S, Kaur P, Ng OT, Garg H TITLE HIV-1 subtype CRF01_AE and B differ in utilization of low levels of CCR5, Maraviroc susceptibility and potential N-glycosylation sites. JOURNAL Virology. 2017 Dec;512:222-233. doi: 10.1016/j.virol.2017.09.026. Epub 2017 Oct 9. PUBMED 29020646 REFERENCE 2 (bases 1 to 8471) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Virology."; date: "2017-12"; volume: "512:222-233. doi"; pages: " 10.1016/j.virol.2017.09.026. Epub 2017 Oct 9" FEATURES Location/Qualifiers source 1..8471 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 3969..4010 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 4020..4037 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4060..4077) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4066..4290 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4336..4764 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4423..4442) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4633..4654 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4773..4793) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4778..5107 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4958..5093 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5020..5039 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5174..5968 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5228..5247) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5838..5857 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6144..6265 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6181..6200) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6235..6254 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6314..6330) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6314..6330) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6327..6349) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6338..6354 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6362..6392) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6407..6428 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6545..6562) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6716..7304) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6796..6815) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7475..8335) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8098..8117 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8336..8440) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagcttggt accgagctcg gatccagtac ccttcaccga tcaagcttta ggcatctcct 961 atggcaggaa gaagcggaag caccgacgag gaactcctcg cagcagtaag aatcatcaag 1021 atcttatacc agagcagtaa gtatatgtaa tgacaccttt agaaattagt gcagtagcag 1081 gaattatagt agcattaatc ttagcaatag tagtgtggac tatagtagtt atagaaatta 1141 agaaagtact aagagagaaa aaaatagaaa gattacttaa gagaataaga gagagagcag 1201 aagatagtgg aaatgagagt gagggagata cggatgaatt ggcaaaactt gtggaattgg 1261 gggactttga tccttgggtt ggggataatt tgtagtgcct cagacaactt gtgggttaca 1321 gtttattatg gggttcctgt gtggaaagat gcagacacca ccctattttg tgcatcagat 1381 gcccaagcac ataagacaga agtgcacaat gtctgggcca cacatgcctg tgtacccaca 1441 gaccccaacc cacaagaaat gctcctggaa aatgtaacag aaaattttaa catgtggaaa 1501 aataacatgg tagagcagat gcatgaagat gtaatcagtt tatgggatca aagtctaaag 1561 ccatgtgtaa agttaactcc tctctgtgtt actttagatt gtaccagtgc taatctgagt 1621 aacagttcca ctagcagagt agcaagcata ataggaaatg taacagagga agtaagaaac 1681 tgctctttta atatgaccac agaactaaat gataagaggc agaagatcca tgcacttttt 1741 tataagcttg atatagtaca aattgatgat aaaaataata atagtgagca ttataggcta 1801 ataaactgta atacttcagt cattaagcag gcttgtccaa aggtatcctt tgatccaatt 1861 cctatacatt attgtactcc agctggttat gcgcttttaa aatgtaatga taagaagttc 1921 aatgggacag ggccatgtag aaatgtcagc tcagtacaat gcacacatgg aattaagcca 1981 gtggtatcaa ctcagttgct gttaaatggt agtctagcag aagatgagat agtaatcaga 2041 agtgaaaatc tcacagacaa tgccaagacc ataatagtgc atcttaataa atctgtagac 2101 atcaattgta ccagaccctc caacaataca agaacaagtg tgtctatagg accaggacga 2161 gtattctata gaacaggaga tataatagga gatataacaa aagcatattg tgagattaat 2221 ggaacaaaat ggaatgaaac tttaacacag gtagaaggaa aattaaagga gcactttaat 2281 aggacaataa tctttcaacc acactcagga ggagatctag agcttacaat gcatcatttt 2341 aattgtagag gggaattttt ctattgcaat acaacaaaac tgtttaatag tccttgcctg 2401 aaaaatgaag aaaatggaac atgtaatagc actatcatac ttccatgtaa gataaagcga 2461 attgtaagat tgtggcagga agtaggtcaa gcaatgtatg ctcctcccat cagcggaatc 2521 attaggtgta actcaactat tacaggaata ctattgacaa gagatggtgg taaccttaat 2581 aacagtacta atgaaaccct cagacctgga ggaggaaata taaaggacaa ttggagaagt 2641 gaattatata aatataaagt agtacaaatt gaaccactag gaatagcacc caccaaggca 2701 aaaagaagag tggtggagag agaaaaaaga gcagtgggaa taggagctat gatctttggg 2761 ttcttaggag cagcaggaag cactatgggc gcggcgtcaa taacgctgac ggtacaggcc 2821 agacaattat tgtctggtat agtgcaacag caaaacaatt tgctgagggc tatagaggcg 2881 cagcagcatc tgttgcaact cacagtctgg ggcattaaac agctccaggc aagagtcctg 2941 gctgtggaaa gatacctaaa agatcaaaag ttcctaggac tttggggctg ctctgggaaa 3001 accatctgca ccactgctgt gccctggaac gacacttgga gtaataaatc tttagaagag 3061 atttgggaca acatgacatg gatagaatgg gagagagaaa ttagcaatta cacaaaccaa 3121 atatatgaga tacttacaga atcgcagaac cagcaggaaa ggaatgaaaa ggatctgtta 3181 gaattggata aatgggcaag tctgtggaat tggtttagca taacaaattg gctgtggtat 3241 ataaggatgt ttataataat agtaggaagt ttaataggtt taagaataat ttttgctgtg 3301 ctttctatag taaatagagt taggcaggga tactcacctt tgtctttcca gatcccttcc 3361 catcatcaga gggaacccga caggcccgaa gaaatcgaag aaggaggtgg cgagcaaggc 3421 agagccagat ccgtgcgctt agtgagcgga ttcttaccgc ttatctggga cgatctacgg 3481 agcctgtgcc tcttcagcta ccaccgcttg agagacttca tcttgatcac aacgaggact 3541 gtggaacttc tgggacacag cagtctcaag ggactgagac gggggtggga aggcctcaaa 3601 tatctgggga atcttctatt atattggggt caggaactaa aaactagtgc catttctttg 3661 tttgatgctg cagcaatagt aatagcggga gggacagata gggttataga agtagcacaa 3721 agagcttgga gagccattct ccacatacct agaagaatca gacagggctt tgaaagggct 3781 ttgctataac atgggaaaca agtggtcaaa aacatggcct cgggtcaggg acagattaga 3841 gaagactcct ccagcagcag atggagtagg agcagtatct caagacggat ccagctaagg 3901 gtcaagacaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgc 3961 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc 4021 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt 4081 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc 4141 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt 4201 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca 4261 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct 4321 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 4381 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 4441 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 4501 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 4561 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 4621 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 4681 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 4741 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa 4801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 4861 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 4921 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 4981 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 5041 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 5101 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg 5161 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 5221 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 5281 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 5341 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 5401 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 5461 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 5521 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 5581 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 5641 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 5701 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 5761 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 5821 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 5881 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 5941 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc 6001 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 6061 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 6121 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 6181 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 6241 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 6301 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 6361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 6421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 6481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 6541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 6601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 6661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 6721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 6781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 6841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 6901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 6961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 7021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 7081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 7141 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 7201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 7261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 7321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 7381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 7441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 7501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 7561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 7621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 7681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 7741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 7801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 7861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 7921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 7981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 8041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 8101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 8161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 8221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 8281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 8341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 8401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 8461 cacctgacgt c //

hKCNQ5(P369T).ires.GFP-pcDNA3

204365

Heterologous expression in mammalian cell lines or Xenopus oocyte (T7 RNA pol)

Aguan Wei

Wei et al J Neurophysiol. 2022 Jul 1;128(1):40-61. doi: 10.1152/jn.00509.2021. Epub 2022 May 18.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/404012/2a472a07-275c-4c9b-b9ec-a3346aa67505/addgene-plasmid-204365-sequence-404012.gbk

full

LOCUS sequence-404012- 10235 bp DNA circular SYN 27-AUG-2024 DEFINITION Heterologous expression in mammalian cell lines or Xenopus oocyte (T7 RNA pol). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10235) AUTHORS Wei AD, Wakenight P, Zwingman TA, Bard AM, Sahai N, Willemsen MH, Schelhaas HJ, Stegmann APA, Verhoeven JS, de Man SA, Wessels MW, Kleefstra T, Shinde DN, Helbig KL, Basinger A, Wagner VF, Rodriguez-Buritica D, Bryant E, Millichap JJ, Millen KJ, Dobyns WB, Ramirez JM, Kalume FK TITLE Human KCNQ5 de novo mutations underlie epilepsy and intellectual disability. JOURNAL J Neurophysiol. 2022 Jul 1;128(1):40-61. doi: 10.1152/jn.00509.2021. Epub 2022 May 18. PUBMED 35583973 REFERENCE 2 (bases 1 to 10235) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1152/jn.00509.2021"; journalName: "J Neurophysiol"; date: "2022-07-1- 1"; volume: "128"; issue: "1"; pages: "40-61" FEATURES Location/Qualifiers source 1..10235 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(36..55) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 282..302 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin complement(311..739) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(421..442) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 633..652 /label=F1ori-R /note="F1 origin, reverse primer" polyA_signal 785..1009 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 998..1015 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" promoter 1035..1053 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1035..1052 /label=SP6 /note="SP6 promoter, forward primer" CDS complement(1069..1788) /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS complement(1072..1788) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind 1463..1482 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1722..1743 /label=EGFP-N /note="EGFP, reverse primer" misc_feature 1789..2375 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1962..1981) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind 2191..2208 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" 5'UTR 5879..5921 /label=Xenopus globin 5'-UTR /note="translational enhancer from the 5'-UTR of the major beta-globin gene of Xenopus laevis" primer_bind complement(5959..5978) /label=T7 /note="T7 promoter, forward primer" promoter complement(5960..5978) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(6023..6226) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(6052..6072) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 6227..6606 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 6778..6797 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 6872..6976 /gene="bla" /label=AmpR promoter CDS 6977..7837 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(7195..7214) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8008..8596 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8497..8516 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8750..8767 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8884..8905 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8920..8950 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8958..8974 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 8963..8985 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8982..8998 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8982..8998 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 9047..9168 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9058..9077) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 9112..9131 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(9342..10136) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(9453..9472) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 10063..10082 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(join(10203..10235,1..297)) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin join(10217..10235,1..117) /label=SV40 ori /note="SV40 origin of replication" ORIGIN 1 tcactacttc tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa 61 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg 121 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct 181 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct 241 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt 301 ccacagaatt aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 361 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 421 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 481 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 541 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 601 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 661 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 721 gcgccgctac agggcgcgtg gggatacccc ctagagcccc agctggttct ttccgcctca 781 gaagccatag agcccaccgc atccccagca tgcctgctat tgtcttccca atcctccccc 841 ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag acaatgcgat 901 gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa 961 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt cgaggctgat 1021 cagcgagctc tagcatttag gtgacactat agaatagggc cctctagatt acttgtacag 1081 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc 1141 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg 1201 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac 1261 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt 1321 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat 1381 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc 1441 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg 1501 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc 1561 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg 1621 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc 1681 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac 1741 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg ttgtggccat 1801 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt 1861 tttttaacct cgactaaaca catgtaaagc atgtgtaccg aggccccaga tcagatccca 1921 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggagagcc 1981 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg 2041 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt 2101 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa 2161 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc 2221 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg 2281 gcaatatggt ggaaaataac atatagacaa acgcacaccg gccttattcc aagcggcttc 2341 ggccagtaac gttagggggg ggggagggag aggggcggat cccgggcgct aaatcaccaa 2401 aagcacacaa attgacattc aaattacatt aataataaaa tgactgaata ggaatgggtt 2461 ggcaaatata tcttgaaata agcagccatt tctattattc cccaattctg ctttagttga 2521 taccttatta gcagctgttg atagccattg tgtttaaata tacattatgg cctaatcttg 2581 aaaatgtagt ttgtatttag tcacaaagca gaaatgcctg aaatttgaag gtgtttactc 2641 ccataatttc aaaatacagt gtcaaatctt ctaaatgacc taaacttgac atattcatac 2701 accccatagc attagctgtg tttggtttct caggcttttt aactcaaggt acattactaa 2761 atgagtgggt taatgtcgat gcaccttgga attttagcca tccctaaaca tgcattttca 2821 tgcaattaaa aggtaacggg cttataaact gcctttcatg ttcttcccga ttcttgcaat 2881 ttcaactttt tagaagggct ttcgaaatag ttcatgcaat gatatgtatg gctaaagaac 2941 tgctatgcct ggaaagaaaa tgaagaactt atttcagttt gacatgaggc aagctgaggg 3001 catctgtact ttctcctgcc ttacaaatgc tctgagatga tcgtgacctt ccagtcctta 3061 gagagtctga tgcaaaggca gcttccctgg caggctgcgg tgcggcatca aaagtgtctg 3121 tctctgtctc ttcgggaccc acctcttcat cagttataaa caatttggat tccctccatt 3181 tggggtaaaa atcttggctg cctctggagc cacttgactc actccctgaa agttgtatat 3241 tcagttcctc ggtcgacctg atcaggtttt gcacagacaa agatttgccc aagtccttcg 3301 gcaccatggg acagacagac aacagagttt ctcctcccat gtcaaagctt ttcctcatag 3361 aacggtcctt ggtgagattt gactgtgcaa cctgaacatt ttccttggag gcaacaaggc 3421 aggtggtgac gtcagaaatg ctttcctgta agcctgcagg gttagggtgc agagtttctg 3481 gcctgggcag atgcttgatg gctgggagag gaggtgggat ctgtaaagtt gttggggctg 3541 ctggcttggg tgccgtattt atttggtttg caatggtgtt ggtggctgcc actgctgagc 3601 catcgctttg actaattggc acctgtgttg cttgactgtg catagtaggg ctaagcgcgt 3661 agaaagtctg ggcactgaac tcatttggcg tcagaatgaa ctgcaggcct ctcgagatgt 3721 tggcactagt tgatctggat aagcagccac tgttttgtgc ggaacccgaa agatctttgc 3781 tatccacagg gctttgatag tcagatgtct gttcacattc aaaaggtggg atctggaatg 3841 aagccaaagc gagggctgag gcagagcctt tccgaaggac ctgttgatag atgtctagta 3901 ggcagtccag cttggattct atggactgta cctgtttttc aaccttgacc acccgaccga 3961 gcatactgag atcgtctgtg gtctcatgtt ctgctgttat tttctctcgg ctcttcttat 4021 ctgatgtgat ttgccctttt ccaagaattt gatcaacacg tgtttgaagg cttttaattc 4081 tacacaacat gtccagatga ccagcagaat attgttcaat gacatctttt acatcatatg 4141 gacgtaatgt ttccttaaac ttccgttttg caacatgaaa tttcataatt ctgatagctc 4201 gaatgacagt tttaagtggt ggggtgaggt cttccactga tacatcacac tggcatcctt 4261 tttcatcata tacatcatca gtgccaaggg ctgtgtcagc atctatcact ggttttggct 4321 gagaactttt gaggcgcagc gagggccgga agcgggttcg gtcgttgaag ctccagctct 4381 tctgcacttt ggtgggactg ccctcggctg tgatgtcggt gcttggggac ctcctgtcac 4441 ctactgaggc ttgtcggctc ttaatactct ggcccctggg gctagccatg cgcactcgct 4501 ccttaaaact tagcttctga cgcaatgtcc actgtcccca actgcttgat gcttcccctt 4561 gttctttctt ggtagggctg caggtgtgca aggccttcaa gtgtggcttc caggttgcaa 4621 tggaaacaga tttctcatca gctgcgtaac tacgccaaac acactgaatg aggttggcag 4681 ctgtgttcct tcttttctca aagtgtttct ggcggtgttg ttcttgtact tttaatgcaa 4741 aacctgagcc aagaatgccg gcaggaagtg caaagaaaga aatgccaagg agtgcaaagc 4801 ctgcagaaag caatcttccc agccaagtta ggggagtttt gtctccatag ccaatagttg 4861 tcaatgtaat tgtgccccac cagagagcat ctgcatatgt agaaaactct ttattggcat 4921 ccttttccac cagatagaca aggaaagacg aaaaaataag aaccaaaaat cctatgtacc 4981 aagctgtgat taattccttg ctgtgagcat aaaccactga acccagtaat ttccaagtgc 5041 ctccccttcg gtccatgcgc accatgcgga ggatctgtag gaaacggaga cttctgagtg 5101 cagacgtggc aaaaatatta ccctgagttt ttgcagaaac aactgctatt gaagcgataa 5161 gaacaatggt atctataaca cagaagggct ttcgagcaaa cctcagtctt ccttgccatc 5221 ctctatatcg acaacagcaa cccgcagacc agattcgaat gatgaactcc aaaccaaaga 5281 cgacaatcat cacgaactcc aggatcaaga ggcaacttga ggccaatttt gtgtgctcag 5341 ggatggtaga aaacactgac aaaatcaagc aaccaaagac aaggagaaaa acgaaagcgt 5401 ggtagatgaa cgcccagccg cggggtctct ccagcacgtt gtacaggtag ttctgcaccc 5461 gccggtactt gacgttgcgc cggcagctct ggctactcgt gtaagagagc ggcttcccca 5521 gcaggctcat ccgggccccc tgcttgcccc ggcggctctc cctcaggcca ccgccgccgc 5581 caccaagcgt ggccgcgcgg gtgcccagca gtagcaggcc gtcgcccctg gcggctgccg 5641 agttcagcag caccctgccc cggcccgact ccacatcctt catgccgctg cccaagcgcc 5701 ccccgcccgc cgccgccgcc gctgcgccgc tcttcaccca gagcccggcg gcgccgccct 5761 cctctcctcc cgcgtggtgg cggggcatgg catctagagc ggccgccagt gtgatggata 5821 tctgcagaat tccagcacac tggcggccgt tactagtgga tccgagctcg gtacccttct 5881 tgttcttttt gcagaagctc agaataaacg ctcaactttg gcagagctag cgtcgacggt 5941 atcgataagc ttgggtctcc ctatagtgag tcgtattaat ttcgataagc cagtaagcag 6001 tgggttctct agttagccag agagctctgc ttatatagac ctcccaccgt acacgcctac 6061 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt 6121 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt 6181 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga 6241 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat 6301 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat 6361 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat 6421 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg 6481 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca 6541 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc 6601 aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa 6661 gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg 6721 cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg 6781 catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc 6841 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 6901 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 6961 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 7021 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 7081 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 7141 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 7201 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 7261 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 7321 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 7381 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 7441 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 7501 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 7561 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 7621 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 7681 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 7741 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 7801 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 7861 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 7921 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 7981 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 8041 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 8101 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 8161 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 8221 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 8281 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 8341 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 8401 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 8461 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 8521 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 8581 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 8641 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 8701 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 8761 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 8821 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 8881 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc 8941 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca 9001 tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag atacattgat 9061 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt 9121 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg ggtgggcgaa 9181 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat 9241 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt 9301 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga 9361 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag 9421 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc 9481 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt 9541 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg 9601 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg 9661 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga 9721 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt 9781 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc 9841 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca 9901 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt 9961 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac 10021 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat 10081 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga 10141 aacgatcctc atcctgtctc ttgatcagat ccgaaaatgg atatacaagc tcccgggagc 10201 tttttgcaaa agcctaggcc tccaaaaaag cctcc //

pBad33_ISCysp13_reRNA

205338

Bacterial expression of ISCysp13 TnpB and its reRNA

Haoyi Wang

Xiang et al Nat Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/411018/f55d192e-0fc3-483a-8aef-6c00163935dc/addgene-plasmid-205338-sequence-411018.gbk

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LOCUS sequence-411018- 5607 bp DNA circular SYN 27-AUG-2024 DEFINITION Bacterial expression of ISCysp13 TnpB and its reRNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5607) AUTHORS Xiang G, Li Y, Sun J, Huo Y, Cao S, Cao Y, Guo Y, Yang L, Cai Y, Zhang YE, Wang H TITLE Evolutionary mining and functional characterization of TnpB nucleases identify efficient miniature genome editors. JOURNAL Nat Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x. PUBMED 37386294 REFERENCE 2 (bases 1 to 5607) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x." FEATURES Location/Qualifiers source 1..5607 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 19..53 /label=BBa_J23108 /note="Anderson constitutive promoter (https://parts.igem.org/Promoters/Catalog/Anderson). Measured strength relative to BBa_J223100: 0.51" RBS 69..80 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" primer_bind complement(1653..1670) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1823..1909 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2001..2028 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2047..2138 /gene="bla" /label=AmpR promoter rep_origin 2596..3051 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2683..2702) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2893..2914 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(3607..4266) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 4181..4200 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(4267..4369) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" primer_bind complement(4761..4778) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4895..5440) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(4975..4994) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5516..5535 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 tgacagctta tcatcgatct gacagctagc tcagtcctag gtataatgct agccccggga 61 aaagatctaa agaggagaaa gaattcatgg ctactaaaag aatcacgttt agactttacc 121 caaataaaga acaaaatgag aaacttcatt attggagacg tttacacaaa gatttataca 181 atgcttgtgt agttaatcga aaaactcaat acaaaaagtt tggtaaatcc attaattact 241 ttgaccaaca aaactcatta cctgagttta aaaaagattg gccagaatac aaagaccttg 301 gtagccatgc ccttcaagca actgtaaaac gggttgattt tgctttccaa cgttttttca 361 agattaaatc aggttatccg aaactaaagt cttcaagaag atataaaggg tggacttatc 421 ctgataaagc tggttggaaa atagacagta atggacatca tggaaagttg acactttcta 481 attttggaac aataaagatg aggggtaaag caagagattg gggaacacca aatacttgca 541 ctatcatctt taaacaagga aaatggtacg cttctgttac tgttagttgt gatccaaaac 601 gagaaacagg taaagaggct tgtggtatcg attttgggac tcatcatgca cttgcttttg 661 acgatggcac tgttattgat aatcctagat ttctcaaaca atcacaggat aaaatcaaca 721 aaacagcaaa gaaatctaga aagaaacgtg caggaaaaaa gggagtaaaa gggtctagaa 781 gatggaaaaa agctgttaga aaaatcggta agattcaatc tttagttggt cgtcaaagac 841 aaaactggca tcatcaagtt tccacacaaa tagttagctg taatagcttg gtggcaactg 901 agaaattaaa cctaaaagga atgaccaaaa aagctaaaag aggtagcaaa agaaaacgtc 961 aaaagactgg attaaatcgg tcaattcttg acgttggaat tggtaactta aaatcattaa 1021 taaaaagcaa agttaccgaa gcaggtgggt tttacattga ggttcctact cgaaaagtta 1081 agccatctca gacttgtcct aattgtggtc atcaaaagaa aaagacatta gcagaacgcc 1141 agcacaactg cgaaaagtgc ctttatagtt gcgacagaga tgttgcatca gcccaggtgt 1201 gtttaaactg ggcaaggggg caggaactgt cctctttaga cgtagagtcg cctagctcta 1261 ctttatgcgg aagcatgagg caactagggg cgaaaaagca caacaatgca aagaagcgtc 1321 agaaacatct agctcaacaa agtggctgag agatctttga cagctagctc agtcctaggt 1381 ataatagcct ttatagttgc gacagagatg ttgcatcagc ccaggtgtgt ttaaactggg 1441 caagggggca ggaactgtcc tctttagacg tagagtcgcc tagctctact ttatgcggaa 1501 gcatgaggca actaggggcg aaaaagcaca acaatgcaaa gaagcgtcag aaacatctag 1561 ctcaacaaag tggctagatg tagttcatgc tcggagatca tcattgcgtt ttttaagctt 1621 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga 1681 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc 1741 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg 1801 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt 1861 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc 1921 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac 1981 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca 2041 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 2101 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 2161 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 2221 tggtgaaagt aaaagatgct gaagatcagt tgggtgcagc aaactattaa ctggcgaact 2281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 2341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 2401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 2461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 2521 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 2581 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 2641 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 2701 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 2761 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg 2821 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 2881 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct 2941 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 3001 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc 3061 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 3121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 3181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 3241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 3301 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 3361 cctacatacc tcgctctgct aatcctgtta ccagtcaggc atttgagaag cacacggtca 3421 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac 3481 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg 3541 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta 3601 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct 3661 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac 3721 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat 3781 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa 3841 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc 3901 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga 3961 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac 4021 cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag 4081 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc 4141 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc 4201 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt 4261 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag 4321 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt 4381 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc 4441 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc 4501 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta 4561 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg 4621 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag 4681 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga 4741 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg 4801 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt 4861 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca 4921 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat 4981 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta 5041 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc 5101 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc 5161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca 5221 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta 5281 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt 5341 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt 5401 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca 5461 gataaaatat ttctaggctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga 5521 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc 5581 gtccggcgta gaggatctgc tcatgtt //

UKNP1.11.4

97380

Expression of HCV E1E2 genes cloned from patient serum. United Kingdom Nottingham Panel x.xx.xx (genotype.patient number.clone number)

Jonathan Ball

Urbanowicz et al J Virol. 2015 Dec 23;90(7):3288-301. doi: 10.1128/JVI.02700-15.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/189796/6b909de2-7f5a-4f31-8bcf-0945892ff4ca/addgene-plasmid-97380-sequence-189796.gbk

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LOCUS sequence-189796- 7250 bp DNA circular SYN 26-AUG-2024 DEFINITION Expression of HCV E1E2 genes cloned from patient serum. United Kingdom Nottingham Panel x.xx.xx (genotype.patient number.clone number). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7250) AUTHORS Urbanowicz RA, McClure CP, Brown RJ, Tsoleridis T, Persson MA, Krey T, Irving WL, Ball JK, Tarr AW TITLE A Diverse Panel of Hepatitis C Virus Glycoproteins for Use in Vaccine Research Reveals Extremes of Monoclonal Antibody Neutralization Resistance. JOURNAL J Virol. 2015 Dec 23;90(7):3288-301. doi: 10.1128/JVI.02700-15. PUBMED 26699643 REFERENCE 2 (bases 1 to 7250) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1128/JVI"; journalName: "J Virol"; date: "2015-12-23- 23"; volume: "90"; issue: "7"; pages: "3288-301" FEATURES Location/Qualifiers source 1..7250 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(45..64) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 236..615 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 616..819 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 770..790 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 864..883 /label=T7 /note="T7 promoter, forward primer" promoter 864..882 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2749..2790 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2800..2817 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2840..2857) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2846..3070 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3116..3544 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3203..3222) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3413..3434 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3553..3573) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3558..3887 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3738..3873 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3800..3819 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3954..4748 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4008..4027) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4618..4637 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4924..5045 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4961..4980) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5015..5034 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5094..5110) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5094..5110) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5107..5129) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5118..5134 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5142..5172) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5187..5208 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5325..5342) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5496..6084) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5576..5595) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6255..7115) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6878..6897 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7116..7220) /gene="bla" /label=AmpR promoter ORIGIN 1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat 61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat 241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat 301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt 661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac 721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc 781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc 841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag 901 ttaagcttgg taccgagctc ggatccagta cccttcacca tgggttgctc tttctctatc 961 ttccttctgg ccctgctctc ttgcttgact gtacccgctt cggcctacca agtgcgcaac 1021 agctcggggc tttaccatgt caccaatgat tgccctaact cgagtattgt gtacgaggcg 1081 gccgatgcca tcctgcacac tccggggtgt gtcccttgcg ttcgcgaggg taacgcctcg 1141 aggtgttggg tggcggtgac ccccacggtg gccaccaggg atggcaaact ccccacaacg 1201 cagcttcgac gtcacatcga tctgcttgtc gggagcgcca ccctctgctc ggccctctac 1261 gtgggggacc tgtgcgggtc tgtctttctt gtcggccaac tgtttacctt ctctcccaga 1321 caccactgga cgacgcaagg ctgcaattgt tctatctatc ccggccacat aacgggtcat 1381 cgcatggcat gggatatgat gatgaactgg tctcctacgg cggcgttggt gatggctcag 1441 ctgctccgga tcccacaagc catcatggac atgatcgctg gtgcccactg gggtgtcctg 1501 gcgggcatag cgtatttctc catggtgggg aactgggcga aagtcctggt agtgctgctg 1561 ctatttgccg gcgtcgacgc ggaaacccac gtcaccggag gaagcgccgg ccacaccgtg 1621 gctgggcttg ctggtttcct cacacgaggc cccaagcaga acatccaact gatcaacacc 1681 aacggcagtt ggcacatcaa tagcacggcc ttgaactgca atgacagcct taacaccggc 1741 tggatagcag gcctcttcta tcatcacaaa ttcaactctt caggctgttc cgaaaggttg 1801 gccagctgcc gacgccttac cgattttgcc cagggctggg gccctatcag ttatgccaac 1861 gggagcggcc ccgacgagcg cccctactgc tggcactacc ccccaagacc ttgcggtatt 1921 gtgcccgcaa agagcgtgtg tggcccggta tattgtttca ctcccagccc cgtggtggtg 1981 ggaacgaccg acaggtcggg cgcgcctacc tacagctggg gtgcaaatga tacggacgtc 2041 ttcgtcctta acaacaccag gccaccgctg ggcaactggt tcggttgcac ctggatgaac 2101 gcaactggat tcaccaaagt gtgcggagcg cccccttgcg tcatcggagg ggcgggcgcc 2161 aacaccttgc gctgccccac tgattgtttc cgcaagcatc cggaagcaac atactctcgg 2221 tgcggctccg gtccctggat tacacccagg tgcatggtcc actatccgta taggctttgg 2281 cattatcctt gtaccatcaa ctacaccata ttcaaagtca ggatgtacgt gggaggggtc 2341 gagcacaggc tggaagctgc ctgcaactgg acgcggggcg aacgctgtga tctggaagac 2401 agggacaggt ccgagctcag cccattgttg ctgtccacca cgcagtggca agtccttccg 2461 tgctctttca cgaccctgcc agccttgtcc actggcctca tccacctcca ccagaacatc 2521 gtggacgtgc agtacttata cggggtaggg tcaagcattg cgtcctgggc cattaaatgg 2581 gagtacgtcg ttctcctgtt cctcctgctt gcagacgcgc gcgtctgctc ctgcttgtgg 2641 atgatgttac tcatatccca agcggaggcg taataaaagg gtcaagacaa ttctgcagat 2701 atccagcaca gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc 2761 cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga 2821 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 2881 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 2941 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 3001 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 3061 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg 3121 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3181 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3241 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 3301 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 3361 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3421 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 3481 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3541 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 3601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3661 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3721 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3781 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3841 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3901 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3961 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 4021 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 4081 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 4141 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 4201 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 4261 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 4321 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 4381 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 4441 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 4501 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 4561 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4621 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4681 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4741 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca 4801 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4861 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4921 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4981 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 5041 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 5101 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 5161 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 5221 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 5281 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 5341 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 5401 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 5461 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 5521 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 5581 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5641 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 5701 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 5761 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5821 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5881 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 5941 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 6001 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 6061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 6121 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 6181 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 6241 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 6301 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 6361 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 6421 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 6481 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 6541 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 6601 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 6661 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 6721 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6781 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6841 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6901 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6961 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 7021 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 7081 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 7141 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 7201 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt //

pAAV9-hSyn-iCytSnFR_ER

177752

Develop a family of genetically encoded fluorescent biosensors for nicotinic agonists, termed iDrugSnFRs, thus enabling optical subcellular pharmacokinetics for nicotinic agonists

Henry Lester

Nichols et al Elife. 2022 Jan 4;11. pii: 74648. doi: 10.7554/eLife.74648.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/349067/a716ee2d-fbf5-4a76-9bd2-71b28f0975f6/addgene-plasmid-177752-sequence-349067.gbk

full

LOCUS sequence-349067- 5910 bp DNA circular SYN 26-AUG-2024 DEFINITION Develop a family of genetically encoded fluorescent biosensors for nicotinic agonists, termed iDrugSnFRs, thus enabling optical subcellular pharmacokinetics for nicotinic agonists. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5910) AUTHORS Nichols AL, Blumenfeld Z, Fan C, Luebbert L, Blom AE, Cohen BN, Marvin JS, Borden PM, Kim CH, Muthusamy AK, Shivange AV, Knox HJ, Campello HR, Wang JH, Dougherty DA, Looger LL, Gallagher T, Rees DC, Lester HA TITLE Fluorescence activation mechanism and imaging of drug permeation with new sensors for smoking-cessation ligands. JOURNAL Elife. 2022 Jan 4;11. pii: 74648. doi: 10.7554/eLife.74648. PUBMED 34982029 REFERENCE 2 (bases 1 to 5910) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; date: "2022-01-4"; volume: "11. pii: 74648. doi"; pages: " 10.7554/eLife.74648" FEATURES Location/Qualifiers source 1..5910 /mol_type="other DNA" /organism="synthetic DNA construct" repeat_region 76..205 /label=AAV2 ITR promoter 287..734 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" primer_bind 1261..1282 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1385..1406) /label=EGFP-N /note="EGFP, reverse primer" misc_feature 2423..3011 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2476..2496) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2894..2905) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3014..3030) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3015..3031) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 3060..3181 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3071..3090) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3125..3144 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3343..3472 /label=AAV2 ITR rep_origin complement(3709..4297) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3789..3808) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4468..5328) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5091..5110 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5329..5433) /gene="bla" /label=AmpR promoter rep_origin complement(join(5459..5910,1..4)) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5596..5617) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5808..5827 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 61 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 121 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 181 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct 241 acgtagccat gctctaggaa gatctctgca gagggccctg cgtatgagtg caagtgggtt 301 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg 361 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa 421 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc 481 ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg 541 ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc 601 cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct 661 gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt 721 gcctgagagc gcagtcgaat tcggcttggg gatatccacc atgctgctat ccgtgccgct 781 gctgctcggc ctcctcggcc tggccatcgc cagatctgcg aacgacaccg tagttgtggg 841 ctcgatcatc tttacagaag ggattatcgt cgcaaacatg gtggcagaga tgattgaggc 901 gcatacagac cttaaggtgg ttcgcaaact gaaccttggc gggtacaacg ttaactttga 961 agccattaaa cgcggaggtg cgaataatgg tattgacatt tacgtggagt acactgggca 1021 cggtcttgtg gatattctgg ggttcccgga gccgaacgtc tatatcaccg ccgacaagca 1081 gaagaacggc atcaaggcga acttcaagat ccgccacaac gtggaggacg gcagcgtgca 1141 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 1201 caaccactac ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca 1261 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 1321 caagggcggt accggaggga gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc 1381 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg 1441 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 1501 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 1561 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 1621 ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa 1681 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 1741 cggcaacatc ctggggcaca agctggagta caactttccg ccgcccgcga ctactgatcc 1801 agaaggtgca tacgaaaccg tgaagaagga gtacaaacgt aaatggaata ttgtatggct 1861 caaaccactg ggattcaaca atacgtatac gcttaccgtt aaagacgaac tggcgaaaca 1921 gtataacctt aaaaccttca gtgacttagc gaaaatctcg gataagctga ttctgggtgc 1981 aacgatgtgg tttttagaag ggcccgatgg ttacccaggc ctgcaaaaac tgtacaattt 2041 caaattcaag cacaccaaaa gcatggacat gggtattcgc tataccgcca ttgataataa 2101 cgaagttcag gtaattgatg cctgggccac tgatggcttg ctggtgagcc acaaattaaa 2161 aattctggag gatgataaag cgttcttccc gccgtattat gctgccccca tcatccgtca 2221 ggatgtctta gataagcatc ctgaactgaa ggacgtgctg aacaaactcg cgaatcaaat 2281 ttcactggaa gaaatgcaga aactgaatta caaggtggac ggtgagggtc aggacccagc 2341 gaaagtagct aaggagtttt tgaaagagaa aggtttaatt ctgcagacag cagaaaaaga 2401 tgagttgtga aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac 2461 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 2521 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 2581 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 2641 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 2701 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 2761 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc 2821 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 2881 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 2941 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 3001 cgcctccccg catcgatacc gtcgacctcg acccgggcgg ccgcttcgag cagacatgat 3061 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat 3121 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt 3181 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt 3241 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg 3301 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg 3361 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg 3421 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta 3481 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 3541 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 3601 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 3661 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 3721 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 3781 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 3841 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 3901 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 3961 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 4021 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 4081 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 4141 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 4201 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 4261 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 4321 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 4381 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 4441 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 4501 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 4561 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 4621 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 4681 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 4741 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 4801 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 4861 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 4921 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 4981 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 5041 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 5101 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 5161 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 5221 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 5281 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 5341 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa 5461 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt 5521 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata 5581 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac 5641 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa 5701 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc 5761 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg 5821 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca 5881 cccgccgcgc ttaatgcgcc gctacagggc //

pCMV-GFP-CENPBpA-fixed-length-clone-8

120186

Randomly picked clone 8 of the pCMV-GFP-CENPBpA-fixed-length library

David Bartel

Wu et al Cell. 2017 May 18;169(5):905-917.e11. doi: 10.1016/j.cell.2017.04.036.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/274598/0716a339-8259-4312-b739-57df8d4251f9/addgene-plasmid-120186-sequence-274598.gbk

full

LOCUS sequence-274598- 4734 bp DNA circular SYN 26-AUG-2024 DEFINITION Randomly picked clone 8 of the pCMV-GFP-CENPBpA-fixed-length library. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4734) AUTHORS Wu X, Bartel DP TITLE Widespread Influence of 3'-End Structures on Mammalian mRNA Processing and Stability. JOURNAL Cell. 2017 May 18;169(5):905-917.e11. doi: 10.1016/j.cell.2017.04.036. PUBMED 28525757 REFERENCE 2 (bases 1 to 4734) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cell.2017.04"; journalName: "Cell"; date: "2017-05-18- 18"; volume: "169"; issue: "5"; pages: "905-917.e11" FEATURES Location/Qualifiers source 1..4734 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 351..554 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 505..525 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 577..657 /label=MCS /note="multiple cloning site" regulatory 659..668 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 665..1384 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 665..1381 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(710..731) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(971..990) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1318..1339 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1726..1745) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 1791..1846 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(1845..1864) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(2207..2223) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2207..2223) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2220..2242) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2231..2247 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2255..2285) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2300..2321 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2380..2575 /label=SV40 promoter /note="SV40 early promoter" rep_origin 2426..2561 /label=SV40 ori /note="SV40 origin of replication" polyA_signal 2581..2715 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2618..2637) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2672..2691 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(2783..2800) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2954..3542) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3034..3053) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3713..4573) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4336..4355 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4574..4678) /gene="bla" /label=AmpR promoter enhancer join(4709..4734,1..350) /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 61 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt 361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 541 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct 601 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc 661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct 721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac 781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc 841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat 901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat 961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac 1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg 1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa 1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct 1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa 1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat 1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa 1381 gtaaagcggc cgcactcctc aggtgcatgc ctctcctccc tctctggcag ggccccatcc 1441 tgggcagagg ggcctggggc tgggcccaga gtccagccgt ccagctgctc ctttcccagt 1501 ttgatttcaa taaatctgtc cactccccct tttgtggggg tggacgtttt aacagccaag 1561 ggtgcatcct tcatggtctg ggcttgcgtc tgtcttgggg gacttattcg tcctggctct 1621 ctttggtcct tgctctggtg ggacatggag gcaagtgttg aaggctgcct atcagaaggt 1681 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc 1741 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa 1801 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata 1861 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata 1921 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa 1981 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt 2041 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt 2101 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct 2161 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc 2221 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 2281 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 2341 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac 2401 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 2461 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 2521 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct 2581 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 2641 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 2701 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 2761 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 2821 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 2881 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 2941 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 3001 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 3061 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 3121 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 3181 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 3241 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 3301 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 3361 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 3421 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 3481 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 3541 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 3601 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 3661 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 3721 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 3781 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 3841 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 3901 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 3961 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 4021 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 4081 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 4141 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 4201 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 4261 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 4321 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 4381 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 4441 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 4501 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 4561 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 4621 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 4681 catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtaa tagt //

PARIS/ZNF746 scFv [N196/16]

206723

Mammalian Expression of PARIS/ZNF746 scFV. Derived from hybridoma N196/16 scFv.

James Trimmer

Mitchell et al Sci Rep. 2023 Sep 27;13(1):16200. doi: 10.1038/s41598-023-43233-4.

partial

GBH-2aG4-gcryBFP.1

188013

Cloning vector for conditional, intronic, targeted integration of a genebreak cassette optimized for zebrafish

Karl Clark

Clark Lab Gene Break Homology Plasmids 2022 (unpublished)

https://media.addgene.org/snapgene-media/v2.0.0/sequences/381108/c45cd4a7-19f8-445e-a928-f53e9a77b8f2/addgene-plasmid-188013-sequence-381108.gbk

full

LOCUS sequence-381108- 7313 bp DNA circular SYN 27-AUG-2024 DEFINITION Cloning vector for conditional, intronic, targeted integration of a genebreak cassette optimized for zebrafish. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7313) TITLE Clark Lab Gene Break Homology Plasmids 2022 REFERENCE 2 (bases 1 to 7313) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..7313 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 278..299 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 314..344 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 352..368 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 357..379 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 376..392 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 376..392 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 388..738 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTCHTSLLDNGPI TLNNANYLKLLS" promoter 413..431 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter complement(479..497) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(507..524) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(507..523) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(516..538) /label=M13/pUC Forward /note="In lacZ gene" protein_bind complement(735..768) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." CDS 946..1002 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 1009..1446 /codon_start=1 /gene="Saccharomyces cerevisiae GAL4 (truncated)" /product="DNA binding domain of the GAL4 transcriptional activator" /label=GAL4 DNA binding domain /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS" CDS 3532..3552 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3556..4257 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" polyA_signal 4267..4491 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" polyA_signal 4561..4642 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" polyA_signal 4766..4847 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(4996..5029) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 5071..5104 /label=attB /bound_moiety="phage phi-C31 integrase" /note="minimal attB site for the phi-C31 integrase (Groth et al., 2000)" promoter 5284..5302 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5578..5597) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 5589..6398 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(5656..5675) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 6541..7129 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7030..7049 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7283..7300 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 tttaaatggt accgagcaga ggtgtaaaaa gtactcaaaa attttactca agtgaaagta 61 caagtactta gggaaaattt tactcaatta aaagtaaaag tatctggcta gaatcttact 121 tgagtaaaag taaaaaagta ctccattaaa attgtacttg agtattaagg aagtaaaagt 181 aaaagcaaga aagaaaacta gagtggtctc cctttagtga gggtcaattg atgggaggcg 241 ttcgggccac agcggacacg caggtgcgca acgcaattaa tgtgagttag ctcactcatt 301 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg 361 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc 421 ctcactaaag ggaacaaaag ctggaggacc tcgagggggg gcccggtacc caattcgccc 481 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa 541 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 601 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 661 tggacctgcc acacttcctt gctagacaat gggcccataa ctttaaataa tgccaattat 721 ttaaagttac tatcataact tcgtatagca tacattatac gaagttatac ctaggcttac 781 cacccactag cggtcagact gcagattgca gcacgaaaca ggaagctgac tccacatggt 841 cacatgctca ctgaagtgtt gacttccctg acagctgtgc actttctaaa ccggttttct 901 cattcattta cagttcagcc ggaattcacc cgggtaccgg aaggtgccac gaacttctct 961 ctgttaaagc aagcaggaga cgtggaagaa aaccccggtc cttctagaaa gctactgtct 1021 tctatcgaac aagcatgcga tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa 1081 ccgaagtgcg ccaagtgtct gaagaacaac tgggagtgtc gctactctcc caaaaccaaa 1141 aggtctccgc tgactagggc acatctgaca gaagtggaat caaggctaga aagactggaa 1201 cagctatttc tactgatttt tcctcgagaa gaccttgaca tgattttgaa aatggattct 1261 ttacaggata taaaagcatt gttaacagga ttatttgtac aagataatgt gaataaagat 1321 gccgtcacag atagattggc ttcagtggag actgatatgc ctctaacatt gagacagcat 1381 agaataagtg cgacatcatc atcggaagag agtagtaaca aaggtcaaag acagttgact 1441 gtatcgtcga ggtcgacccc gggaatacag atctctcgag ccgccccccc gaccgatgtc 1501 agcctggggg acgagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg 1561 ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc 1621 ccccacgact ccgcccccta cggcgctctg gatatgtaac tagtaattaa gtctcagcca 1681 ccgttaactg aacatgtcaa aacctgtgga gactgttgag atttgatgtt ctgaaaagat 1741 aaagcctata aataaaatgt tgcccaaatt tcctgcctga tgtttttctt tgtctttgct 1801 acatggcttt gctgctcgga tcggctcact ctgtgtatgc cacgttcact ttgtactctc 1861 cttctcacgg taggtttatt atttttagat gtgcagttag tttctgtgaa ataacacacc 1921 acacactgat attgtctgtg cattgacttg gtgagtgcac attgtttttg atcttgacat 1981 atttatattt gattgatcag gtgaactgtg tgaatctaaa gtgctccata cagatgttct 2041 gcattgaaaa tattctcatt ttattagtgg aagtgagtgt atgccacatc caatcaattt 2101 cagcaaacac cccagtatga tttaatgcaa aaaaatgaag gtatcaaaca cgcattacta 2161 ctttgcagtt aaatatttaa catttattcc aacacgaaaa aaagcagtaa ataacacttt 2221 gacaaacacg tcaggacatc ttatttttgt caccctcaca ggcaatttag tataatatat 2281 tatatatata tatatatcat ataataatat tcagtataat atatatatat atatcatatt 2341 ataatattca gtataatata aaacacaaac acatatatgt ataatataat ataacatttt 2401 tatttattga gatgcctcta tggaccgtgt tataagaagt aaagatcagg agaagtaaac 2461 atgaagtgta attatgaata ctgatgttaa attaagctat gatgagtttt cactgttaat 2521 ttaccatctc aattaaatgt tgatgcctcc atgaccaagt taagcagatg agactgagga 2581 caactgtaga agacaagatg ttcactttgc tgaatatagc tggcttgaca gttatctatg 2641 actctataaa tatatatata tttttttttt tataaaatga tttatttata actatatatc 2701 catttctcag acaggtgctt catatccctc actcccgtag ctgtccatgc tggatctgtc 2761 cccgttgttt ttaaaaagct aaataagtta ttaacatgac tgcatccagc gagccaaacc 2821 tgtctggtgt acagctacca gagaagcttg agatcctagt caccggcgtt cctagttctt 2881 taaactgtta caaagtgttt ggaccagcaa ctgaattgca ctgtaaatta ccacatcagg 2941 gtctccgggg tcaacgctgc taagcaatgg actggaaaat ttgtgacatt gagtttttgg 3001 aataactggg aatgctggat aactctttgg gtaaagctat ggaacaaaag taccagcatg 3061 aactaagaaa tgactgtcac agcacaacat caaacacaat gcaagacaaa ggttctctgc 3121 ttttgcctta cttgacagaa aaagtaagct ctaggttcat cgcatgcaga cagcagcaat 3181 acatgacata tataaaggct ggctcttttc tcccttccat tgaactgaaa cttccactca 3241 gtcagacttg cgggggatcc acagcctgac taactcgaca aaccccccca aacctaaggt 3301 gagttgatct ttaagctttt tacattttca gctcgcatat atcaattcgc gcgcattgtc 3361 acactagcac ccactagcgg tcagactgca gattgcagca cgaaacagga agctgactcc 3421 acatggtcac atgctcactg aagtgttgac ttccctgaca gctgtgcact ttctaaaccg 3481 gttttctcat tcatttacag ttcagccgga attcacctcg agaccatggc tccaaagaag 3541 aagcgtaagg taaccatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg 3601 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac 3661 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc 3721 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc 3781 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca 3841 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc 3901 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag 3961 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa 4021 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag 4081 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg 4141 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag 4201 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct caattaaagc 4261 tgagatctgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 4321 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 4381 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 4441 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 4501 cggaaagaac caaagggcga attaattcat tgatgagttt ggacaaacca caactagaat 4561 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat 4621 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca 4681 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtcattgatg 4741 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg 4801 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt 4861 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa 4921 acctctacaa atgtggtcat tcagatcaat taaccctcac ccgggctagc gatgcatggg 4981 agctcaagac gtctgataac ttcgtatagc atacattata cgaagttatc tacctaactt 5041 taaataatgc caattattta aagttacttc cgcgcccggg gagcccaagg gcacgccctg 5101 gcacctgacg tacgttgtct agcaaggaag tgaagagcta tccggatata tatgtatgtt 5161 atcataactg tactggctat gtcaggtggt aatgactgta acgttacgtt actcgttgta 5221 ggcacgacat tgaatatgct cttctccgct gtggcccgaa cgcctcccgc tagcgatggg 5281 atctaatacg actcactata gggagaccaa gggaaaatag aatgaagtga tctccaaaaa 5341 ataagtactt tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa 5401 aaaatgtaat taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa 5461 atccccaaaa ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc 5521 tctgggatcc agctgcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa 5581 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg 5641 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa 5701 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta 5761 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc 5821 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg 5881 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata 5941 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc 6001 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt 6061 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga 6121 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct 6181 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag 6241 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt 6301 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata 6361 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt 6421 aacactggca aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 6481 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6541 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6601 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6661 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 6721 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6781 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6841 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6901 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgg ttcccgaagg 6961 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 7021 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 7081 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 7141 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 7201 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 7261 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaac aga //

TFORF1558

141839

Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens.

Feng Zhang

Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/415133/aacc491a-ba29-4e72-a192-17d0c0adefbe/addgene-plasmid-141839-sequence-415133.gbk

full

LOCUS sequence-415133- 8719 bp DNA circular SYN 26-AUG-2024 DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8719) AUTHORS Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F TITLE A transcription factor atlas of directed differentiation. JOURNAL Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. PUBMED 36608654 REFERENCE 2 (bases 1 to 8719) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5"; volume: "186"; issue: "1"; pages: "209-229.e26" FEATURES Location/Qualifiers source 1..8719 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 5..231 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 232..412 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 459..584 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1077..1310 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1495..1539 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 1770..1980 /label=SV40 promoter /note="SV40 early promoter" promoter 1792..1988 /label=SV40 promoter /note="SV40 early promoter" rep_origin 1839..1974 /label=SV40 ori /note="SV40 origin of replication" CDS 2020..2622 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS" primer_bind complement(2020..2039) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2516..2536 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 2665..2782 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2920..4098 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3151..4089 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4046..4066 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 4109..4118 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" misc_feature 4576..5164 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4629..4649) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5047..5058) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5236..5469 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5547..5668 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5584..5603) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5638..5657 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5708..5843 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5863..5882) /label=T7 /note="T7 promoter, forward primer" promoter complement(5864..5882) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5892..5909) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5892..5908) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5901..5923) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6050..6505 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6137..6156) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6347..6368 /label=F1ori-F /note="F1 origin, forward primer" promoter 6531..6635 /gene="bla" /label=AmpR promoter CDS 6636..7496 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6854..6873) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7667..8255 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8156..8175 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8409..8426 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8543..8564 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8579..8609 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8617..8633 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 8622..8644 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8641..8657 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8641..8657 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8676..8696 /label=T3 /note="T3 promoter, forward primer" promoter 8678..8696 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa 61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac 121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa 181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc 241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa 1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca 1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg 2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag 2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc 2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca 2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg 2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc 2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg 2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc 3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc 3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt 3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct 3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac 3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct 3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc 3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt 3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg 3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt 3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga 3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc 3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg 3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg 3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt 3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt 3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg 3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt 4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt 4081 ccatttcagg tgtcgtgagg ctagcaccgc caccatggga gacaagaaga gccccaccag 4141 gccgaagcgg cagccgaagc cgtcctcgga tgagggttac tgggactgta gcgtctgcac 4201 cttccggaac agcgccgagg ccttcaagtg catgatgtgc gatgtgcgga agggcacctc 4261 cacccgggac agcaaggaag gggggaagct ggtgtcctac tccacagcca gtcttggggt 4321 tagaggaacc ctgagaaata gagtaggtgg tggcagctca gaagagaaga aacaggctga 4381 atacctggca cctggaagaa gaaggaatat agtacacagg ggagttggcc caggacagag 4441 aagtgggcct agtttaaaag aggcttagta atgaactagt ccgtaacttg aaagtatttc 4501 gatttcttgg ctttatatat cttgtggaaa ggacgagtca ctcaagagta ggttggtggt 4561 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 4621 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 4681 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 4741 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 4801 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 4861 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 4921 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc 4981 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 5041 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 5101 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 5161 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa 5221 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg 5281 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag 5341 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc 5401 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa 5461 tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa 5521 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat 5581 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 5641 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac 5701 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 5761 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 5821 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat 5881 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 5941 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 6001 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt 6061 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 6121 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 6181 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 6241 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 6301 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 6361 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 6421 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 6481 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc 6541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 6601 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 6661 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 6721 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 6781 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 6841 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac 6901 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 6961 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 7021 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 7081 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 7141 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc 7201 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 7261 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 7321 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc 7381 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 7441 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 7501 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 7561 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 7621 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 7681 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 7741 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 7801 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 7861 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 7921 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 7981 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 8041 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 8101 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 8161 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 8221 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 8281 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 8341 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 8401 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 8461 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 8521 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 8581 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 8641 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac 8701 aaaagctgga gctgcaagc //

pDONR223_IKZF5_p.E135K

81384

Gateway Donor vector containing IKZF5 , part of the Target Accelerator Plasmid Collection.

Kim et al Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/167855/aa1b8818-a217-48fe-8288-658de2d9aa46/addgene-plasmid-81384-sequence-167855.gbk

full

LOCUS sequence-167855- 4056 bp DNA circular SYN 26-AUG-2024 DEFINITION Gateway Donor vector containing IKZF5 , part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4056) AUTHORS Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn WC TITLE Systematic Functional Interrogation of Rare Cancer Variants Identifies Oncogenic Alleles. JOURNAL Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4. PUBMED 27147599 REFERENCE 2 (bases 1 to 4056) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: "2016-07"; volume: "6"; issue: "7"; pages: "714-26" FEATURES Location/Qualifiers source 1..4056 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 43..65 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 57..74 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 58..74 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 90..189 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1456..1555) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1572..1591) /label=T7 /note="T7 promoter, forward primer" promoter complement(1573..1591) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1596..1612) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1596..1612) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2043..2834 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 2927..3515 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3416..3435 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3669..3686 /label=L4440 /note="L4440 vector, forward primer" terminator complement(3845..3872) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(3964..4050) /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta 61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga 121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa 181 aagttggcac catgggtgaa aaaaaaccag agcctttgga cttcgtgaaa gattttcagg 241 aatacctgac tcagcagacc catcacgtga acatgatttc tggatcagtt agtggggaca 301 aagaagcaga ggctcttcag ggagctggaa cagatggtga tcaaaatgga cttgatcacc 361 catctgttga agtttccttg gatgaaaact caggaatgtt agtagacggg tttgaaagga 421 cctttgatgg gaagcttaag tgtcggtact gcaactatgc cagcaaagga acagcccggc 481 ttattgaaca catcagaatc cacacaggtg aaaaacctca tcgatgtcat ctttgtccat 541 ttgcatctgc ttatgagcgt catctggaag cccatatgcg ttctcatact ggaaaaaaac 601 catacaaatg tgaattatgt tccttccgct gcagtgatcg aagtaacttg tcccatcatc 661 gaaggcgcaa gcataaaatg gtaccaatta aaggtactag gtcttcctta agcagcaaga 721 aaatgtgggg ggttttacag aagaaaacaa gcaatctggg ctatagcaga agagcactaa 781 tcaacttaag tccaccttcc gtggtggttc agaaaccaga ctaccttaac gattttaccc 841 acgaaatccc aaatatccag actgactcct atgaaagtat ggcaaaaacc acaccaactg 901 gtggccttcc aagggacccc caagaactca tggttgataa ccctttgaat cagctctcga 961 ctctagcagg gcagttgtcc agtctgccac ccgaaaacca aaaccctgca tcacctgatg 1021 tagttccctg ccctgatgaa aagcctttca tgattcagca gccctctacc caagcagtag 1081 tttctgccgt atcagcaagt attcctcaga gctcctctcc cacaagccca gaacctcggc 1141 catcccatag tcaaaggaac tatagtccag tggcaggtcc aagcagtgag ccaagtgccc 1201 acacgagcac tcccagcata ggaaacagcc agccaagcac cccagcccca gccctgccgg 1261 tccaggaccc tcagcttctg caccactgcc agcactgtga tatgtacttt gcagacaaca 1321 tcctttacac tattcatatg ggatgtcatg ggtatgaaaa tccttttcag tgtaatatat 1381 gtggatgcaa atgtaaaaac aagtatgatt ttgcctgtca ttttgcaaga gggcacataa 1441 ccaacattga ttgattgcca actttcttgt acaaagttgg cattataaga aagcattgct 1501 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc 1561 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct 1621 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc 1681 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt 1741 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc 1801 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac 1861 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg 1921 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga 1981 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca 2041 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca 2101 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg 2161 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg 2221 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga 2281 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc 2341 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag 2401 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag 2461 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac 2521 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg 2581 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg 2641 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt 2701 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg 2761 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg 2821 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt 2881 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 2941 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 3001 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 3061 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 3121 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 3181 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 3241 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 3301 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 3361 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 3421 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 3481 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 3541 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 3601 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 3661 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 3721 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 3781 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa 3841 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg 3901 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg 3961 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca 4021 gtcttccgac tgagcctttc gttttatttg atgcct //

pEGFP-SUN1_916

125850

Low

Expresses SUN1_916 tagged with GFP in mammalian cells

Miki Hieda

Nishioka et al Nucleus. 2016 Nov;7(6):572-584. doi: 10.1080/19491034.2016.1260802.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/433864/6795e473-ead6-420b-9a55-0b8741f02f81/addgene-plasmid-125850-sequence-433864.gbk

full

LOCUS sequence-433864- 7457 bp DNA circular SYN 25-AUG-2024 DEFINITION Expresses SUN1_916 tagged with GFP in mammalian cells. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7457) AUTHORS Nishioka Y, Imaizumi H, Imada J, Katahira J, Matsuura N, Hieda M TITLE SUN1 splice variants, SUN1_888, SUN1_785, and predominant SUN1_916, variably function in directional cell migration. JOURNAL Nucleus. 2016 Nov;7(6):572-584. doi: 10.1080/19491034.2016.1260802. PUBMED 27858498 REFERENCE 2 (bases 1 to 7457) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1080/19491034.2016"; journalName: "Nucleus"; date: "2016-11"; volume: "7"; issue: "6"; pages: "572-584" FEATURES Location/Qualifiers source 1..7457 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 3397..3406 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 3403..4122 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3403..4119 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3448..3469) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3709..3728) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4056..4077 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4245..4366 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4282..4301) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4336..4355 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4373..4828) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4510..4531) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4722..4741 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4855..4959 /gene="bla" /label=AmpR promoter promoter 4961..5318 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 4961..5152 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(4984..5004) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5169..5304 /label=SV40 ori /note="SV40 origin of replication" CDS 5353..6147 /codon_start=1 /product="aminoglycoside phosphotransferase" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS 5353..6147 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5407..5426) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6017..6036 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6335..6354) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6379..6426 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6755..7343 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7244..7263 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctgcccat ggatttttct cggcttcaca tgtacagtcc tccccagtgt 661 gtgccggaga acacgggcta cacgtatgcg ctcagttcca gctattcttc agatgctctg 721 gattttgaga cggagcacaa attggaccct gtatttgatt ctccacggat gtcccgccgt 781 agtttgcgcc tggccacgac agcatgcacc ctgggggatg gtgaggctgt gggtgccgac 841 agcggcacca gcagcgctgt ctccctgaag aaccgagcgg ccagaacaac aaaacagcgc 901 agaagcacaa acaaatcagc ttttagtatc aaccacgtgt caaggcaggt cacgtcctct 961 ggcgtcagcc acggcggcac tgtcagcctg caggatgctg tgactcgacg gcctcctgta 1021 ttggacgagt cttggattcg tgaacagacc acagtggacc acttctgggg tcttgatgat 1081 gatggtgatc ttaaaggtgg aaataaagct gccattcagg gaaacgggga tgtgggagcc 1141 gccgccgcca ccgcgcacaa cggcttctcc tgcagcaact gcagcatgct gtccgagcgc 1201 aaggacgtgc tcacggcgca ccccgcggcc cccgggcccg tgtcgagagt ttattctagg 1261 gacaggaatc aaaaatgcgg tgcgtctttc tacgtgaata ggattttgtg gctggccaga 1321 tacactgcat catctttttc atcattttta gttcaacttt ttcaagtggt tttaatgaag 1381 ctcagttatg aatcagaaaa ttacaaattg aaaactcatg aatcaaaaga ttgtgaatcc 1441 gaaagctata agtcaaaaag ccatgaatca aaagctcatg ccagttacta tgggagaatg 1501 aatgtgagag aggttctcag agaggatggc cacctcagtg taaatgggga agcgctgtgc 1561 gacgactgta agggcaagag gcacctcgac gcgcacacag ccgcccactc gcagtcgcca 1621 cggctgcccg gtcgggcagg gaccctctgg cacatctggg catgtgcagg ttacttcttg 1681 ctgcagattc tgcgcaggat cggagctgtg ggccaggctg tgtccaggac ggcgtggtcg 1741 gccctttggc tggccgtggt tgctccaggg aaggcagcct ctggagtgtt ctggtggctg 1801 gggattggat ggtaccagtt tgttactttg atttcttggc tgaatgtgtt tcttcttacc 1861 aggtgccttc gaaacatctg caagttttta gtcttgctca tcccactctt ccttttacta 1921 gcaggtctct ccttacgggg ccagggcaat ttcttttcgt tcttgcccgt gttgaactgg 1981 gcaagcatgc atagaacaca gcgggtggat gacccccagg acgtgtttaa acccacgact 2041 tctcgcctga agcagcctct gcagggtgac agtgaggctt ttccgtggca ttggatgagt 2101 ggcgtggagc agcaggtggc ctctctgtct ggacagtgcc accaccatgg tgagaatctc 2161 cgagagctga ccactttgct acagaagctg caggctcggg tggaccagat ggaaggcggc 2221 gctgccgggc cgtcagcttc ggtcagagac gctgtgggac agcccccgag ggagactgac 2281 tttatggcct ttcaccaaga acatgaagtg cgtatgtcac acttggaaga tattctggga 2341 aaactgagag aaaaatctga ggccatccag aaggaactag aacagaccaa gcaaaaaaca 2401 atcagtgcgg ttggtgagca gctcctgccc acagtcgagc acctccagct ggagctggat 2461 cagctaaagt cagagctgtc cagctggcga cacgtgaaga ccggctgtga gacagtggat 2521 gccgtacaag aaagagtgga cgtgcaagtc agagaaatgg tgaaactcct gttttccgaa 2581 gatcagcaag gcggttctct ggaacagctg ctgcagaggt tctcatcaca gtttgtgagc 2641 aaaggcgact tgcagacgat gctgcgagac ctgcagctgc agatcctgcg gaacgtcacc 2701 caccacgttt ccgtgaccaa gcagctccca acctcagaag ccgtggtgtc tgctgtgagc 2761 gaggcggggg cgtctggaat aacagaggcg caagcacgtg ccatcgtgaa cagcgccttg 2821 aagctgtatt cccaagataa gaccgggatg gtggactttg ctctggaatc tggtggtggc 2881 agcatcttga gtactcgctg ttctgaaact tacgaaacca aaacggcgct gatgagtctg 2941 tttgggatcc cgctgtggta cttctcgcag tccccgcgcg tggtcatcca gcctgacatt 3001 taccccggta actgctgggc atttaaaggc tcccaggggt acctggtggt gaggctctcc 3061 atgatgatcc acccagccgc cttcactctg gagcacatcc ctaagacgct gtcgccaaca 3121 ggcaacatca gcagcgcccc caaggacttc gccgtctatg gattagaaaa tgagtatcag 3181 gaagaagggc agcttctggg acagttcacg tatgatcagg atggggagtc gctccagatg 3241 ttccaggccc tgaaaagacc cgacgacaca gctttccaaa tagtggaact tcggattttt 3301 tctaactggg gccatcctga gtatacctgt ctgtatcggt tcagagttca tggcgaacct 3361 gtcaaggtcg acggtaccgc gggcccggga tccatcgcca ccatggtgag caagggcgag 3421 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac 3481 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag 3541 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc 3601 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag 3661 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac 3721 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg 3781 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac 3841 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc 3901 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac 3961 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc 4021 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc 4081 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga 4141 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc 4201 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag 4261 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4321 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta 4381 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac 4441 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg 4501 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa 4561 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt 4621 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt 4681 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga 4741 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc 4801 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga 4861 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 4921 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc 4981 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 5041 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 5101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 5161 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 5221 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt 5281 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga 5341 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 5401 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 5461 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 5521 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 5581 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 5641 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 5701 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 5761 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 5821 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 5881 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 5941 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 6001 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 6061 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 6121 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 6181 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 6241 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 6301 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg 6361 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt 6421 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac 6481 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg 6541 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt 6601 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 6661 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 6721 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 6781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 6841 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 6901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 6961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 7021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 7081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 7141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 7201 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 7261 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 7321 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 7381 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 7441 gtattaccgc catgcat //

pLCMV:ECFP(loxP)(FRT)HNF1B_P328L329del

30528

Gerhart Ryffel

Roose et al Transgenic Res. 2009 Aug . 18(4):595-605.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/14099/0721d5b1-77d6-4630-ba5e-e1f3e127ada1/addgene-plasmid-30528-sequence-14099.gbk

full

LOCUS sequence-14099-e 7376 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7376) AUTHORS Roose M, Sauert K, Turan G, Solomentsew N, Werdien D, Pramanik K, Senkel S, Ryffel GU, Waldner C TITLE Heat-shock inducible Cre strains to study organogenesis in transgenic Xenopus laevis. JOURNAL Transgenic Res. 2009 Aug . 18(4):595-605. PUBMED 19266305 REFERENCE 2 (bases 1 to 7376) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Transgenic Res. 2009 Aug . 18(4):595-605." FEATURES Location/Qualifiers source 1..7376 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind complement(34..67) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 75..108 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" regulatory 118..127 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 124..843 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(169..190) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(430..449) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 777..798 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 957..1091 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(1099..1132) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 1140..1173 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" CDS 1202..1231 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1241..1270 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1280..1309 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1319..1348 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1358..1387 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1421..1450 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" intron 3258..3323 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" polyA_signal 3370..3504 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3658..3678) /label=T3 /note="T3 promoter, forward primer" promoter complement(3658..3676) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3697..3713) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3697..3713) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3710..3732) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3721..3737 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3745..3775) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3790..3811 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3928..3945) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4099..4687) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4179..4198) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4858..5718) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5481..5500 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5719..5823) /gene="bla" /label=AmpR promoter rep_origin complement(5849..6304) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5986..6007) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6198..6217 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6392..7376 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttataactt cgtatagcat acattatacg 61 aagttataac tagtgaagtt cctattctct agaaagtata ggaacttcag atccatcgcc 121 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 181 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 241 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 301 accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg 361 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 421 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 481 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 541 cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag 601 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 661 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 721 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 781 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 841 taatctagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa 901 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact 961 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 1021 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 1081 atgtctggat ctgaattcat aacttcgtat agcatacatt atacgaagtt ataggatccg 1141 aagttcctat tctctagaaa gtataggaac ttcccgggcc taggatcgat ttaaagctat 1201 ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga 1261 agaggacttg aatgaaatgg agcaaaagct catttctgaa gaggacttga atgaaatgga 1321 gcaaaagctc atttctgaag aggacttgaa tgaaatggag caaaagctca tttctgaaga 1381 ggacttgaat gaaatggaga gcttgggcga cctcaccatg gagcaaaagc tcatttctga 1441 agaggacttg aattcaagct tccatggtgt ccaagctcac gtcgctccag caagaactcc 1501 tgagcgccct gctgagctcc ggggtcacca aggaggtgct ggttcaggcc ttggaggagt 1561 tgctgccatc cccgaacttc ggggtgaagc tggagacgct gcccctgtcc cctggcagcg 1621 gggccgagcc cgacaccaag ccggtcttcc atactctcac caacggccac gccaagggcc 1681 gcttgtccgg cgacgagggc tccgaggacg gcgacgacta tgacacacct cccatcctca 1741 aggagctgca ggcgctcaac accgaggagg cggcggagca gcgggcggag gtggaccgga 1801 tgctcagtga ggacccttgg agggctgcta aaatgatcaa gggttacatg cagcaacaca 1861 acatccccca gagggaggtg gtcgatgtca ccggcctgaa ccagtcgcac ctctcccagc 1921 atctcaacaa gggcacccct atgaagaccc agaagcgtgc cgctctgtac acctggtacg 1981 tcagaaagca acgagagatc ctccgacaat tcaaccagac agtccagagt tctggaaata 2041 tgacagacaa aagcagtcag gatcagctgc tgtttctctt tccagagttc agtcaacaga 2101 gccatgggcc tgggcagtcc gatgatgcct gctctgagcc caccaacaag aagatgcgcc 2161 gcaaccggtt caaatggggg cccgcgtccc agcaaatctt gtaccaggcc tacgatcggc 2221 aaaagaaccc cagcaaggaa gagagagagg ccttagtgga ggaatgcaac agggcagaat 2281 gtttgcagcg aggggtgtcc ccctccaaag cccacggcct gggctccaac ttggtcactg 2341 aggtccgtgt ctacaactgg tttgcaaacc gcaggaagga ggaggcattc cggcaaaagc 2401 tggccatgga cgcctatagc tccaaccaga ctcacagcct gaacgctctc ccacggctcc 2461 ccccaccacc agcccagctc ctctcctcca aacaagctgt caggagtgcg ctacagccag 2521 cagggaaaca atgagatcac ttcctcctca acaatcagtc accatggcaa cagcgccatg 2581 gtgaccagcc agtcggtttt acagcaagtc tccccagcca gcctggaccc aggccacaat 2641 ctcctctcac ctgatggtaa aatgatctca gtctcaggag gaggtttgcc cccagtcagc 2701 accttgacga atatccacag cctctcccac cataatcccc agcaatctca aaacctcatc 2761 atgacacccc tctctggagt catggcaatt gcacaaagcc tcaacacctc ccaagcacag 2821 agtgtccctg tcatcaacag tgtggccggc agcctggcag ccctgcagcc cgtccagttc 2881 tcccagcagc tgcacagccc tcaccagcag cccctcatgc agcagagccc aggcagccac 2941 atggcccagc agcccttcat ggcagctgtg actcagctgc agaactcaca catgtacgca 3001 cacaagcagg aaccccccca gtattcccac acctcccggt ttccatctgc aatggtggtc 3061 acagatacca gcagcatcag tacactcacc aacatgtctt caagtaaaca gtgtcctcta 3121 caagcctggt gacggacgtc taggagatct caccggtgac tagtcagggg ggatctgcat 3181 ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag 3241 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga 3301 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt 3361 ggaatgccag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga 3421 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc 3481 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt 3541 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct 3601 gattatgatc tgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct 3661 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 3721 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 3781 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 3841 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 3901 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 3961 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4021 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4261 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4501 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 4801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 4861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 4921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 4981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 5041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 5101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 5161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 5221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 5281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 5341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 5401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 5461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 5521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 5581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 5641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 5701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 5761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 5821 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc 5881 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc 5941 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 6001 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc 6061 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa 6121 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg 6181 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt 6241 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc 6301 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 6361 tcgctattac gccagtcgag gcggccgcgt cgaccatagc caattcaata tggcgtatat 6421 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat 6481 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt 6541 ggcaccatgc caattcaata tggcggactt ggcactgtgc caactgggga ggggtctact 6601 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat 6661 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa 6721 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc 6781 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat 6841 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt 6901 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga 6961 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc 7021 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc 7081 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac 7141 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac 7201 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac 7261 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa 7321 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac //

pDONR221-SLC26A3_STOP

161186

Gateway entry clone with codon-optimized ORF sequence for subcloning into custom expression vectors. Contains a STOP codon at the end of the codon-optimized ORF sequence.

Superti-Furga et al Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 10.1038/d41573-020-00056-6.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/339574/9eb15f0b-92cb-4fdc-92a5-0dc02aea197a/addgene-plasmid-161186-sequence-339574.gbk

full

LOCUS sequence-339574- 4845 bp DNA circular SYN 26-AUG-2024 DEFINITION Gateway entry clone with codon-optimized ORF sequence for subcloning into custom expression vectors. Contains a STOP codon at the end of the codon-optimized ORF sequence. . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4845) AUTHORS Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov V, Wolf G, Crawford M, Everley R, ... Steppan CM TITLE The RESOLUTE consortium: unlocking SLC transporters for drug discovery. JOURNAL Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 10.1038/d41573-020-00056-6. PUBMED 32265506 REFERENCE 2 (bases 1 to 4845) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; pages: "429-430" FEATURES Location/Qualifiers source 1..4845 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 1..100 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(2402..2501) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2518..2537) /label=T7 /note="T7 promoter, forward primer" promoter complement(2519..2537) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2542..2558) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2542..2558) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2660..2679) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2671..3480 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(2738..2757) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3627..4215 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4116..4135 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4369..4386 /label=L4440 /note="L4440 vector, forward primer" terminator complement(4545..4572) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(4664..4750) /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 4799..4821 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4813..4830 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4814..4830 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa 61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgat cgagcccttc 121 ggcaaccagt acatcgtggc ccggcccgtg tactctacca acgccttcga ggagaatcac 181 aagaagaccg gcagacacca caagacattt ctggatcacc tgaaggtgtg ctgtagctgc 241 tcccctcaga aggccaagag gatcgtgctg tccctgttcc ctatcgcctc ttggctgcca 301 gcctaccgcc tgaaggagtg gctgctgagc gatatcgtgt ctggcatcag caccggaatc 361 gtggccgtgc tgcagggact ggcctttgcc ctgctggtgg acatcccccc cgtgtacggc 421 ctgtatgcct ccttctttcc agccatcatc tatctgttct ttggcacatc ccggcacatc 481 tctgtgggcc ctttcccaat cctgtctatg atggtgggac tggccgtgag cggagccgtg 541 tccaaggccg tgccagacag aaatgccacc acactgggcc tgcccaacaa tagcaacaat 601 agctccctgc tggacgatga gagggtgcgc gtggcagcag cagcctctgt gaccgtgctg 661 agcggcatca tccagctggc cttcggcatc ctgcggatcg gctttgtggt catctacctg 721 tctgagagcc tgatctccgg ctttaccaca gcagcagcag tgcacgtgct ggtgtctcag 781 ctgaagttca tctttcagct gaccgtgcca agccacacag atcccgtgtc catcttcaag 841 gtgctgtatt ccgtgttttc tcagatcgag aagaccaaca tcgccgacct ggtgacagcc 901 ctgatcgtgc tgctggtggt gagcatcgtg aaggagatca atcagaggtt caaggacaag 961 ctgcccgtgc ctatcccaat cgagtttatc atgaccgtga tcgcagcagg cgtgtcctac 1021 ggctgcgatt tcaagaaccg ctttaaggtg gccgtggtgg gcgacatgaa tcctggcttc 1081 cagccaccca tcacaccaga tgtggagacc ttccagaaca cagtgggcga ctgttttgga 1141 atcgcaatgg tggccttcgc cgtggccttt agcgtggcct ccgtgtattc tctgaagtac 1201 gattatcctc tggacggcaa ccaggagctg atcgccctgg gcctgggcaa tatcgtgtgc 1261 ggcgtgttcc ggggatttgc aggatctacc gccctgtcca gatctgccgt gcaggagagc 1321 accggaggca agacacagat cgcaggactg atcggcgcca tcatcgtgct gatcgtggtg 1381 ctggcaatcg gcttcctgct ggcacccctg cagaagtccg tgctggccgc cctggccctg 1441 ggaaacctga agggcatgct gatgcagttt gccgagatcg gccggctgtg gagaaaggat 1501 aagtacgact gtctgatctg gatcatgacc ttcatcttta caatcgtgct gggactggga 1561 ctgggactgg cagccagcgt ggcctttcag ctgctgacca tcgtgttccg gacacagttt 1621 cctaagtgct ccaccctggc caatatcggc agaacaaaca tctacaagaa taagaaggat 1681 tactacgaca tgtacgagcc agagggcgtg aagatcttca ggtgtccctc tcctatctac 1741 tttgccaaca tcggcttctt tcggagaaag ctgatcgatg ccgtgggctt cagcccactg 1801 cgcatcctgc ggaagagaaa taaggccctg aggaagatcc gcaagctgca gaagcaggga 1861 ctgctgcagg tgaccccaaa gggctttatc tgtaccgtgg acacaatcaa ggattccgac 1921 gaggagctgg ataacaatca gatcgaggtg ctggaccagc caatcaacac cacagatctg 1981 cccttccaca tcgactggaa cgacgatctg cccctgaata tcgaggtgcc taagatcagc 2041 ctgcactccc tgatcctgga tttctctgcc gtgagctttc tggacgtgtc tagcgtgagg 2101 ggcctgaaga gcatcctgca ggagttcatc cgcatcaagg tggacgtgta catcgtgggc 2161 accgacgatg actttatcga gaagctgaat agatatgagt tctttgacgg cgaggtgaag 2221 tcctctatct tctttctgac catccacgat gccgtgctgc acatcctgat gaagaaggac 2281 tacagcacat ccaagttcaa cccctcccag gagaaggatg gcaagatcga ctttaccatc 2341 aacacaaatg gcggcctgag gaatcgcgtg tatgaggtgc ctgtggagac aaagttctaa 2401 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc 2461 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc 2521 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa 2581 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct 2641 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg 2701 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat 2761 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg 2821 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta 2881 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat 2941 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa 3001 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat 3061 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg 3121 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc 3181 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca 3241 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata 3301 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta 3361 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt 3421 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa 3481 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc 3541 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac 3601 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 3661 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 3721 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 3781 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 3841 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 3901 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 3961 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 4021 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 4081 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 4141 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 4201 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 4261 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 4321 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 4381 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 4441 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 4501 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca 4561 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc 4621 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga 4681 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc 4741 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc 4801 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc //

pSin-NE1m

172883

Expresses the genetically-encoded fluorescent norepinephrine(NE) sensor GRAB_NE1m in neurons

J. Julius Zhu

Feng et al Neuron. 2019 May 22;102(4):745-761.e8. doi: 10.1016/j.neuron.2019.02.037. Epub 2019 Mar 25.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/340541/d4ac8883-9bff-4c12-8206-2ba6c64826b0/addgene-plasmid-172883-sequence-340541.gbk

full

LOCUS sequence-340541- 11956 bp DNA circular SYN 26-AUG-2024 DEFINITION Expresses the genetically-encoded fluorescent norepinephrine(NE) sensor GRAB_NE1m in neurons. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11956) AUTHORS Feng J, Zhang C, Lischinsky JE, Jing M, Zhou J, Wang H, Zhang Y, Dong A, Wu Z, Wu H, Chen W, Zhang P, Zou J, Hires SA, Zhu JJ, Cui G, Lin D, Du J, Li Y TITLE A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo Detection of Norepinephrine. JOURNAL Neuron. 2019 May 22;102(4):745-761.e8. doi: 10.1016/j.neuron.2019.02.037. Epub 2019 Mar 25. PUBMED 30922875 REFERENCE 2 (bases 1 to 11956) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.neuron.2019.02.037"; journalName: "Neuron"; date: "2019-05-22- 22"; volume: "102"; issue: "4"; pages: "745-761.e8" FEATURES Location/Qualifiers source 1..11956 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 677..695 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 677..694 /label=SP6 /note="SP6 promoter, forward primer" sig_peptide 8353..8415 /codon_start=1 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader /translation="METDTLLLWVLLLWVPGSTGD" protein_bind 8416..8440 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 9543..9564 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(9670..9691) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(9931..9950) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" promoter 10821..10925 /gene="bla" /label=AmpR promoter CDS 10926..11786 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(11144..11163) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcgagctcg tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat 661 gtatcataca catacgattt aggtgacact atagattgac ggcgtagtac acactattga 721 atcaaacagc cgaccaattg cactaccatc acaatggaga agccagtagt aaacgtagac 781 gtagaccccc agagtccgtt tgtcgtgcaa ctgcaaaaaa gcttcccgca atttgaggta 841 gtagcacagc aggtcactcc aaatgaccat gctaatgcca gagcattttc gcatctggcc 901 agtaaactaa tcgagctgga ggttcctacc acagcgacga tcttggacat aggcagcgca 961 ccggctcgta gaatgttttc cgagcaccag tatcattgtg tctgccccat gcgtagtcca 1021 gaagacccgg accgcatgat gaaatatgcc agtaaactgg cggaaaaagc gtgcaagatt 1081 acaaacaaga acttgcatga gaagattaag gatctccgga ccgtacttga tacgccggat 1141 gctgaaacac catcgctctg ctttcacaac gatgttacct gcaacatgcg tgccgaatat 1201 tccgtcatgc aggacgtgta tatcaacgct cccggaacta tctatcatca ggctatgaaa 1261 ggcgtgcgga ccctgtactg gattggcttc gacaccaccc agttcatgtt ctcggctatg 1321 gcaggttcgt accctgcgta caacaccaac tgggccgacg agaaagtcct tgaagcgcgt 1381 aacatcggac tttgcagcac aaagctgagt gaaggtagga caggaaaatt gtcgataatg 1441 aggaagaagg agttgaagcc cgggtcgcgg gtttatttct ccgtaggatc gacactttat 1501 ccagaacaca gagccagctt gcagagctgg catcttccat cggtgttcca cttgaatgga 1561 aagcagtcgt acacttgccg ctgtgataca gtggtgagtt gcgaaggcta cgtagtgaag 1621 aaaatcacca tcagtcccgg gatcacggga gaaaccgtgg gatacgcggt tacacacaat 1681 agcgagggct tcttgctatg caaagttact gacacagtaa aaggagaacg ggtatcgttc 1741 cctgtgtgca cgtacatccc ggccaccata tgcgatcaga tgactggtat aatggccacg 1801 gatatatcac ctgacgatgc acaaaaactt ctggttgggc tcaaccagcg aattgtcatt 1861 aacggtagga ctaacaggaa caccaacacc atgcaaaatt accttctgcc gatcatagca 1921 caagggttca gcaaatgggc taaggagcgc aaggatgatc ttgataacga gaaaatgctg 1981 ggtactagag aacgcaagct tacgtatggc tgcttgtggg cgtttcgcac taagaaagta 2041 cattcgtttt atcgcccacc tggaacgcag acctgcgtaa aagtcccagc ctcttttagc 2101 gcttttccca tgtcgtccgt atggacgacc tctttgccca tgtcgctgag gcagaaattg 2161 aaactggcat tgcaaccaaa gaaggaggaa aaactgctgc aggtctcgga ggaattagtc 2221 atggaggcca aggctgcttt tgaggatgct caggaggaag ccagagcgga gaagctccga 2281 gaagcacttc caccattagt ggcagacaaa ggcatcgagg cagccgcaga agttgtctgc 2341 gaagtggagg ggctccaggc ggacatcgga gcagcattag ttgaaacccc gcgcggtcac 2401 gtaaggataa tacctcaagc aaatgaccgt atgatcggac agtatatcgt tgtctcgcca 2461 aactctgtgc tgaagaatgc caaactcgca ccagcgcacc cgctagcaga tcaggttaag 2521 atcataacac actccggaag atcaggaagg tacgcggtcg aaccatacga cgctaaagta 2581 ctgatgccag caggaggtgc cgtaccatgg ccagaattcc tagcactgag tgagagcgcc 2641 acgttagtgt acaacgaaag agagtttgtg aaccgcaaac tataccacat tgccatgcat 2701 ggccccgcca agaatacaga agaggagcag tacaaggtta caaaggcaga gcttgcagaa 2761 acagagtacg tgtttgacgt ggacaagaag cgttgcgtta agaaggaaga agcctcaggt 2821 ctggtcctct cgggagaact gaccaaccct ccctatcatg agctagctct ggagggactg 2881 aagacccgac ctgcggtccc gtacaaggtc gaaacaatag gagtgatagg cacaccgggg 2941 tcgggcaagt cagctattat caagtcaact gtcacggcac gagatcttgt taccagcgga 3001 aagaaagaaa attgtcgcga aattgaggcc gacgtgctaa gactgagggg tatgcagatt 3061 acgtcgaaga cagtagattc ggttatgctc aacggatgcc acaaagccgt agaagtgctg 3121 tacgttgacg aagcgttcgc gtgccacgca ggagcactac ttgccttgat tgctatcgtc 3181 aggccccgca agaaggtagt actatgcgga gaccccatgc aatgcggatt cttcaacatg 3241 atgcaactaa aggtacattt caatcaccct gaaaaagaca tatgcaccaa gacattctac 3301 aagtatatct cccggcgttg cacacagcca gttacagcta ttgtatcgac actgcattac 3361 gatggaaaga tgaaaaccac gaacccgtgc aagaagaaca ttgaaatcga tattacaggg 3421 gccacaaagc cgaagccagg ggatatcatc ctgacatgtt tccgcgggtg ggttaagcaa 3481 ttgcaaatcg actatcccgg acatgaagta atgacagccg cggcctcaca agggctaacc 3541 agaaaaggag tgtatgccgt ccggcaaaaa gtcaatgaaa acccactgta cgcgatcaca 3601 tcagagcatg tgaacgtgtt gctcacccgc actgaggaca ggctagtgtg gaaaaccttg 3661 cagggcgacc catggattaa gcagctcact aacataccta aaggaaactt tcaggctact 3721 atagaggact gggaagctga acacaaggga ataattgctg caataaacag ccccactccc 3781 cgtgccaatc cgttcagctg caagaccaac gtttgctggg cgaaagcatt ggaaccgata 3841 ctagccacgg ccggtatcgt acttaccggt tgccagtgga gcgaactgtt cccacagttt 3901 gcggatgaca aaccacattc ggccatttac gccttagacg taatttgcat taagtttttc 3961 ggcatggact tgacaagcgg actgttttct aaacagagca tcccactaac gtaccatccc 4021 gccgattcag cgaggccggt agctcattgg gacaacagcc caggaacccg caagtatggg 4081 tacgatcacg ccattgccgc cgaactctcc cgtagatttc cggtgttcca gctagctggg 4141 aagggcacac aacttgattt gcagacgggg agaaccagag ttatctctgc acagcataac 4201 ctggtcccgg tgaaccgcaa tcttcctcac gccttagtcc ccgagtacaa ggagaagcaa 4261 cccggcccgg tcgaaaaatt cttgaaccag ttcaaacacc actcagtact tgtggtatca 4321 gaggaaaaaa ttgaagctcc ccgtaagaga atcgaatgga tcgccccgat tggcatagcc 4381 ggtgcagata agaactacaa cctggctttc gggtttccgc cgcaggcacg gtacgacctg 4441 gtgttcatca acattggaac taaatacaga aaccaccact ttcagcagtg cgaagaccat 4501 gcggcgacct taaaaaccct ttcgcgttcg gccctgaatt gccttaaccc aggaggcacc 4561 ctcgtggtga agtcctatgg ctacgccgac cgcaacagtg aggacgtagt caccgctctt 4621 gccagaaagt ttgtcagggt gtctgcagcg agaccagatt gtgtctcaag caatacagaa 4681 atgtacctga ttttccgaca actagacaac agccgtacac ggcaattcac cccgcaccat 4741 ctgaattgcg tgatttcgtc cgtgtatgag ggtacaagag atggagttgg agccgcgccg 4801 tcataccgca ccaaaaggga gaatattgct gactgtcaag aggaagcagt tgtcaacgca 4861 gccaatccgc tgggtagacc aggcgaagga gtctgccgtg ccatctataa acgttggccg 4921 accagtttta ccgattcagc cacggagaca ggcaccgcaa gaatgactgt gtgcctagga 4981 aagaaagtga tccacgcggt cggccctgat ttccggaagc acccagaagc agaagccttg 5041 aaattgctac aaaacgccta ccatgcagtg gcagacttag taaatgaaca taacatcaag 5101 tctgtcgcca ttccactgct atctacaggc atttacgcag ccggaaaaga ccgccttgaa 5161 gtatcactta actgcttgac aaccgcgcta gacagaactg acgcggacgt aaccatctat 5221 tgcctggata agaagtggaa ggaaagaatc gacgcggcac tccaacttaa ggagtctgta 5281 acagagctga aggatgaaga tatggagatc gacgatgagt tagtatggat tcatccagac 5341 agttgcttga agggaagaaa gggattcagt actacaaaag gaaaattgta ttcgtacttc 5401 gaaggcacca aattccatca agcagcaaaa gacatggcgg agataaaggt cctgttccct 5461 aatgaccagg aaagtaatga acaactgtgt gcctacatat tgggtgagac catggaagca 5521 atccgcgaaa agtgcccggt cgaccataac ccgtcgtcta gcccgcccaa aacgttgccg 5581 tgcctttgca tgtatgccat gacgccagaa agggtccaca gacttagaag caataacgtc 5641 aaagaagtta cagtatgctc ctccaccccc cttcctaagc acaaaattaa gaatgttcag 5701 aaggttcagt gcacgaaagt agtcctgttt aatccgcaca ctcccgcatt cgttcccgcc 5761 cgtaagtaca tagaagtgcc agaacagcct accgctcctc ctgcacaggc cgaggaggcc 5821 cccgaagttg tagcgacacc gtcaccatct acagctgata acacctcgct tgatgtcaca 5881 gacatctcac tggatatgga tgacagtagc gaaggctcac ttttttcgag ctttagcgga 5941 tcggacaact ctattactag tatggacagt tggtcgtcag gacctagttc actagagata 6001 gtagaccgaa ggcaggtggt ggtggctgac gttcatgccg tccaagagcc tgcccctatt 6061 ccaccgccaa ggctaaagaa gatggcccgc ctggcagcgg caagaaaaga gcccactcca 6121 ccggcaagca atagctctga gtccctccac ctctcttttg gtggggtatc catgtccctc 6181 ggatcaattt tcgacggaga gacggcccgc caggcagcgg tacaacccct ggcaacaggc 6241 cccacggatg tgcctatgtc tttcggatcg ttttccgacg gagagattga tgagctgagc 6301 cgcagagtaa ctgagtccga acccgtcctg tttggatcat ttgaaccggg cgaagtgaac 6361 tcaattatat cgtcccgatc agccgtatct tttcctctac gcaagcagag acgtagacgc 6421 aggagcagga ggactgaata ctgactaacc ggggtaggtg ggtacatatt ttcgacggac 6481 acaggccctg ggcacttgca aaagaagtcc gttctgcaga accagcttac agaaccgacc 6541 ttggagcgca atgtcctgga aagaattcat gccccggtgc tcgacacgtc gaaagaggaa 6601 caactcaaac tcaggtacca gatgatgccc accgaagcca acaaaagtag gtaccagtct 6661 cgtaaagtag aaaatcagaa agccataacc actgagcgac tactgtcagg actacgactg 6721 tataactctg ccacagatca gccagaatgc tataagatca cctatccgaa accattgtac 6781 tccagtagcg taccggcgaa ctactccgat ccacagttcg ctgtagctgt ctgtaacaac 6841 tatctgcatg agaactatcc gacagtagca tcttatcaga ttactgacga gtacgatgct 6901 tacttggata tggtagacgg gacagtcgcc tgcctggata ctgcaacctt ctgccccgct 6961 aagcttagaa gttacccgaa aaaacatgag tatagagccc cgaatatccg cagtgcggtt 7021 ccatcagcga tgcagaacac gctacaaaat gtgctcattg ccgcaactaa aagaaattgc 7081 aacgtcacgc agatgcgtga actgccaaca ctggactcag cgacattcaa tgtcgaatgc 7141 tttcgaaaat atgcatgtaa tgacgagtat tgggaggagt tcgctcggaa gccaattagg 7201 attaccactg agtttgtcac cgcatatgta gctagactga aaggccctaa ggccgccgca 7261 ctatttgcaa agacgtataa tttggtccca ttgcaagaag tgcctatgga tagattcgtc 7321 atggacatga aaagagacgt gaaagttaca ccaggcacga aacacacaga agaaagaccg 7381 aaagtacaag tgatacaagc cgcagaaccc ctggcgactg cttacttatg cgggattcac 7441 cgggaattag tgcgtaggct tacggccgtc ttgcttccaa acattcacac gctttttgac 7501 atgtcggcgg aggattttga tgcaatcata gcagaacact tcaagcaagg cgacccggta 7561 ctggagacgg atatcgcatc attcgacaaa agccaagacg acgctatggc gttaaccggt 7621 ctgatgatct tggaggacct gggtgtggat caaccactac tcgacttgat cgagtgcgcc 7681 tttggagaaa tatcatccac ccatctacct acgggtactc gttttaaatt cggggcgatg 7741 atgaaatccg gaatgttcct cacacttttt gtcaacacag ttttgaatgt cgttatcgcc 7801 agcagagtac tagaagagcg gcttaaaacg tccagatgtg cagcgttcat tggcgacgac 7861 aacatcatac atggagtagt atctgacaaa gaaatggctg agaggtgcgc cacctggctc 7921 aacatggagg ttaagatcat cgacgcagtc atcggtgaga gaccacctta cttctgcggc 7981 ggatttatct tgcaagattc ggttacttcc acagcgtgcc gcgtggcgga tcccctgaaa 8041 aggctgttta agttgggtaa accgctccca gccgacgacg agcaagacga agacagaaga 8101 cgcgctctgc tagatgaaac aaaggcgtgg tttagagtag gtataacagg cactttagca 8161 gtggccgtga cgacccggta tgaggtagac aatattacac ctgtcctact ggcattgaga 8221 acttttgccc agagcaaaag agcattccaa gccatcagag gggaaataaa gcatctctac 8281 ggtggtccta aatagtcagc atagtacatt tcatctgact aataccacaa caccaccacc 8341 tctagatcca ccatggagac agacacactc ctgctatggg tactgctgct ctgggttcca 8401 ggttccactg gtgacacaag tttgtacaaa aaagttggca ccaccggtgg ctccctgcag 8461 ccggacgcgg gcaacgcgag ctggaacggg accgaggcgc cggggggcgg cgcccgggcc 8521 accccttact ccctgcaggt gacgctgacg ctggtgtgcc tggccggcct gctcatgctg 8581 ctcaccgtgt tcggcaacgt gctcgtcatc atcgccgtgt tcacgagccg cgcgctcaag 8641 gcgccccaaa acctcttcct ggtgtctctg gcctcggccg acatcctggt ggccacgctc 8701 gtcatccctt tctcgctggc caacgaggtc atgggctact ggtacttcgg caaggcttgg 8761 tgcgagatct acctggcgct cgacgtgctc ttctgcacgt cgtccatcgt gcacctgtgc 8821 gccatcagcc tggaccgcta ctggtccatc acacaggcca tcgagtacaa cctgaagcgc 8881 acgccgcgcc gcatcaaggc catcatcatc accgtgtggg tcatctcggc cgtcatctcc 8941 ttcccgccgc tcatctccat cgagaagaag ggcggcggcg gcggcccgca gccggccgag 9001 ccgcgctgcg agatcaacga ccagaagtgg tacgtcatct cgtcgtgcat cggctccttc 9061 ttcgctccct gcctcatcat gatcctggtc tacgtgcgca tctaccagat cgccaagcgt 9121 cgcacccgcg tgccacccag ccgccggggt ccggacgccg tcgccgcgcc gccggggggc 9181 accgagcgca ggcccaacgg tctgggcccc gagcgcagcg cgggcccggg gggcgcagag 9241 gccgaaccgc tgcccaccca gctcaacggc gcccctggcg agcccgcgcc ggccgggccg 9301 cgcgacaccg acgcgctgga cctggaggag ggaggaaacg tctatatcaa ggccgacaag 9361 cagaagaacg gcatcaaggc gaacttccac atccgccaca acatcgagga cggcggcgtg 9421 cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 9481 gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat 9541 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg 9601 tacaagggcg gtaccggagg gagcatggtg agaaagggcg aggagctgtt caccggggtg 9661 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 9721 gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 9781 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc 9841 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 9901 tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 9961 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 10021 gaggacggca acatcctggg gcacaagctg gagtacaaca ccggagcagc agcacgctgg 10081 cgcgggcggc agaaccgcga gaagcgcttc acgttcgtgc tggccgtggt catcggagtg 10141 ttcgtggtgt gctggttccc cttcttcttc acctacacgc tcacggccgt cgggtgctcc 10201 gtgccacgca cgctcttcaa attcttcttc tggttcggct actgcaacag ctcgttgaac 10261 ccggtcatct acaccatctt caaccacgat ttccgccgcg ccttcaagaa gatcctctgt 10321 cggggggaca ggaagcggat cgtgttgtaa gcatgcaggc cttgggccca atgatccgac 10381 cagcaaaact cgatgtactt ccgaggaact gatgtgcata atgcatcagg ctggtacatt 10441 agatccccgc ttaccgcggg caatatagca acactaaaaa ctcgatgtac ttccgaggaa 10501 gcgcagtgca taatgctgcg cagtgttgcc acataaccac tatattaacc atttatctag 10561 cggacgccaa aaactcaatg tatttctgag gaagcgtggt gcataatgcc acgcagcgtc 10621 tgcataactt ttattatttc ttttattaat caacaaaatt ttgtttttaa catttcaaaa 10681 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaggggaatt cctcgataat taagcggccg 10741 ctcgagcggc cgcttaatta atcgagggga attaattctt gaagacgaaa gggccaggtg 10801 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 10861 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 10921 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 10981 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 11041 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 11101 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 11161 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 11221 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 11281 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 11341 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 11401 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 11461 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 11521 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 11581 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 11641 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 11701 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 11761 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 11821 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 11881 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 11941 agatcaaagg atcttc //

pFUSB4_TAAG

48562

Stephen Ekker

Ma et al PLoS One. 2013 May 29;8(5):e65259. doi: 10.1371/journal.pone.0065259. Print 2013.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/70262/c3c662b1-23bf-4e08-b77f-56b29bc4eda9/addgene-plasmid-48562-sequence-70262.gbk

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LOCUS sequence-70262-e 2906 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2906) AUTHORS Ma AC, Lee HB, Clark KJ, Ekker SC TITLE High Efficiency In Vivo Genome Engineering with a Simplified 15-RVD GoldyTALEN Design. JOURNAL PLoS One. 2013 May 29;8(5):e65259. doi: 10.1371/journal.pone.0065259. Print 2013. PUBMED 23734242 REFERENCE 2 (bases 1 to 2906) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One."; date: "2013-05-29"; volume: "8(5):e65259. doi"; pages: " 10.1371/journal.pone.0065259. Print 2013" FEATURES Location/Qualifiers source 1..2906 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 8..30 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 22..39 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 23..39 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 858..1649 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 1742..2330 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2231..2250 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2484..2501 /label=L4440 /note="L4440 vector, forward primer" terminator complement(2660..2687) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(2779..2865) /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagcgtct catggcctga 61 ccccggacca agtggtggcc tatcgccagc aacggtggcg gcaagcaagc gctcgaaacg 121 gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg 181 gctatcgcca gcaacattgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg 241 gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacatt 301 ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat 361 ggcctgaccc cggaccaagt ggtggctatc gccagcaaca atggcggcaa gcaagcgctc 421 gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa 481 gtggtggcta tcgagacgtc tagaccagcc aggacagaaa tgcctcgact tcgctgctac 541 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat 601 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca 661 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg 721 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg 781 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta 841 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag 901 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg 961 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga 1021 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg 1081 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg 1141 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca 1201 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc 1261 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg 1321 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact 1381 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt 1441 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc 1501 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag 1561 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag 1621 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta 1681 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 1741 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 1801 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 1861 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 1921 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 1981 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 2041 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 2101 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag 2161 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 2221 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 2281 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 2341 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 2401 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 2461 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa 2521 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt 2581 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg 2641 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg 2701 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt 2761 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat 2821 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc 2881 ctactctcgc gttaacgcta gcatgg //

pDONR223-CLK3

23753

Johannessen et al Nature. 2010 Dec 16;468(7326):968-72. doi: 10.1038/nature09627

https://media.addgene.org/snapgene-media/v2.0.0/sequences/267450/04534c2d-a0d0-456d-8085-94712839292d/addgene-plasmid-23753-sequence-267450.gbk

full

LOCUS sequence-267450- 4260 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4260) AUTHORS Johannessen CM, Boehm JS, Kim SY, Thomas SR, Wardwell L, Johnson LA, Emery CM, Stransky N, Cogdill AP, Barretina J, Caponigro G, Hieronymus H, Murray RR, Salehi-Ashtiani K, Hill DE, Vidal M, Zhao JJ, Yang X, Alkan O, Kim S, Harris JL, Wilson CJ, Myer VE, Finan PM, Root DE, Roberts TM, Golub T, Flaherty KT, Dummer R, Weber BL, Sellers WR, Schlegel R, Wargo JA, Hahn WC, Garraway LA TITLE COT drives resistance to RAF inhibition through MAP kinase pathway reactivation. JOURNAL Nature. 2010 Dec 16;468(7326):968-72. doi: 10.1038/nature09627 PUBMED 21107320 REFERENCE 2 (bases 1 to 4260) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: "Nature"; date: "2010-12-16- 16"; volume: "468"; issue: "7326"; pages: "968-72" FEATURES Location/Qualifiers source 1..4260 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 375..394 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 628..645 /label=L4440 /note="L4440 vector, forward primer" terminator complement(804..831) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(923..1009) /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 1058..1080 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1072..1089 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1073..1089 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1105..1204 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(2675..2774) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2791..2810) /label=T7 /note="T7 promoter, forward primer" promoter complement(2792..2810) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2815..2831) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2815..2831) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 3262..4053 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin join(4146..4260,1..474) /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 61 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 541 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 601 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 661 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 721 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc 781 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt 841 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa 901 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac 961 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag 1021 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg 1081 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt 1141 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt 1201 ggcatgcatc actgtaagcg ataccgctcc cctgaaccag acccgtacct gagctaccga 1261 tggaagagga ggaggtccta cagtcgggaa catgaaggga gactgcgata cccgtcccga 1321 agggagcctc ccccacgaag atctcggtcc agaagccatg accgcctgcc ctaccagagg 1381 aggtaccggg agcgccgtga cagcgataca taccggtgtg aagagcggag cccatccttt 1441 ggagaggact actatggacc ttcacgttct cgtcatcgtc ggcgatcgcg ggagaggggg 1501 ccataccgga cccgcaagca tgcccaccac tgccacaaac gccgcaccag gtcttgtagc 1561 agcgcctcct cgagaagcca acagagcagt aagcgcagca gccggagtgt ggaagatgac 1621 aaggagggtc acctggtgtg ccggatcggc gattggctcc aagagcgata tgagattgtg 1681 gggaacctgg gtgaaggcac ctttggcaag gtggtggagt gcttggacca tgccagaggg 1741 aagtctcagg ttgccctgaa gatcatccgc aacgtgggca agtaccggga ggctgcccgg 1801 ctagaaatca acgtgctcaa aaaaatcaag gagaaggaca aagaaaacaa gttcctgtgt 1861 gtcttgatgt ctgactggtt caacttccac ggtcacatgt gcatcgcctt tgagctcctg 1921 ggcaagaaca cctttgagtt cctgaaggag aataacttcc agccttaccc cctaccacat 1981 gtccggcaca tggcctacca gctctgccac gcccttagat ttctgcatga gaatcagctg 2041 acccatacag acttgaaacc agagaacatc ctgtttgtga attctgagtt tgaaaccctc 2101 tacaatgagc acaagagctg tgaggagaag tcagtgaaga acaccagcat ccgagtggct 2161 gactttggca gtgccacatt tgaccatgag caccacacca ccattgtggc cacccgtcac 2221 tatcgcccgc ctgaggtgat ccttgagctg ggctgggcac agccctgtga cgtctggagc 2281 attggctgca ttctctttga gtactaccgg ggcttcacac tcttccagac ccacgaaaac 2341 cgagagcacc tggtgatgat ggagaagatc ctagggccca tcccatcaca catgatccac 2401 cgtaccagga agcagaaata tttctacaaa gggggcctag tttgggatga gaacagctct 2461 gacggccggt atgtgaagga gaactgcaaa cctctgaaga gttacatgct ccaagactcc 2521 ctggagcacg tgcagctgtt tgacctgatg aggaggatgt tagaatttga ccctgcccag 2581 cgcatcacac tggccgaggc cctgctgcac cccttctttg ctggcctgac ccctgaggag 2641 cggtccttcc acaccagccg caacccaagc agatgcccaa ctttcttgta caaagttggc 2701 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat 2761 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata 2821 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat 2881 aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg 2941 ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg 3001 acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg 3061 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt 3121 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac 3181 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc 3241 cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta 3301 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg 3361 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg 3421 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact 3481 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac 3541 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag 3601 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc 3661 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc 3721 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg 3781 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt 3841 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg 3901 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga 3961 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg 4021 aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc 4081 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 4141 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 4201 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact //

LI B-C mCherry

54426

Martin Parniske

Binder et al PLoS One. 2014 Feb 13;9(2):e88218. doi: 10.1371/journal.pone.0088218. eCollection 2014.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/84646/27b7b0b7-79f9-4fd2-9b40-b2f0dee99cc2/addgene-plasmid-54426-sequence-84646.gbk

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LOCUS sequence-84646-e 3247 bp DNA circular SYN 26-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3247) AUTHORS Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske M TITLE A modular plasmid assembly kit for multigene expression, gene silencing and silencing rescue in plants. JOURNAL PLoS One. 2014 Feb 13;9(2):e88218. doi: 10.1371/journal.pone.0088218. eCollection 2014. PUBMED 24551083 REFERENCE 2 (bases 1 to 3247) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One."; date: "2014-02-13"; volume: "9(2):e88218. doi"; pages: " 10.1371/journal.pone.0088218. eCollection 2014" FEATURES Location/Qualifiers source 1..3247 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 587..605 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(643..665) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 765..784 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 978..1000 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 992..1009 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 993..1009 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 1095..1802 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1240..1258) /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(1892..1908) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1892..1908) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1905..1927) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1916..1932 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(1940..1970) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1985..2006 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2123..2140) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2294..2882) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2374..2393) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2997..3025 /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" CDS join(3214..3247,1..500) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" ORIGIN 1 gcagtcgccc taaaacaaag ttaggtggct caagtatggg catcattcgc acatgtaggc 61 tcggccctga ccaagtcaaa tccatgcggg ctgctcttga tcttttcggt cgtgagttcg 121 gcgacgtagc cacctactcc caacatcagc cggactccga ttacctcggg aacttgctcc 181 gtagtaagac attcatcgcg cttgctgcct tcgaccaaga agcggttgtt ggcgctctcg 241 cggcttacgt tctgcccagg tttgagcagc cgcgtagtga gatctatatc tatgatctcg 301 cagtctccgg cgagcaccgg aggcagggca ttgccaccgc gctcatcaat ctcctcaagc 361 atgaggccaa cgcgcttggt gcttatgtga tctacgtgca agcagattac ggtgacgatc 421 ccgcagtggc tctctataca aagttgggca tacgggaaga agtgatgcac tttgatatcg 481 acccaagtac cgccacctaa caattcgttc aagccgagat cggcttcccg gccgcggagt 541 tgttcggtaa attgtcacaa cgccgctcat gacattaacc tataaaaata ggcgtatcac 601 gaggcccttt tgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 661 cccggagaag gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 721 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 781 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 841 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 901 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 961 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct 1021 cggtacctcg cgaatgcatc tagatatcgg atcccgggcc cgtcgactgc agaggtacgg 1081 gtctcatctg aacaatggtg agcaagggcg aggaggataa catggccatc atcaaggagt 1141 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg 1201 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg 1261 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg 1321 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct 1381 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact 1441 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct 1501 ccgacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc gagcggatgt 1561 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg 1621 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg 1681 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg 1741 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca 1801 agggtggagg aggttctgga ggcggtggaa gtggtggcgg aggtagcggc acctgagacc 1861 tctgcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 1921 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 1981 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 2041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 2101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 2161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 2221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 2281 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 2341 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 2401 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 2461 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 2521 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 2581 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 2641 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 2701 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 2761 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 2821 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 2881 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 2941 aagggatttt ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga acccagttga 3001 cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt 3061 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt 3121 atgactgttt ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg 3181 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg 3241 cagcagg //

pBabe-kappa-HA-dL5-myc-ADRB2

101253

Expresses HA-dL5(E52D)-cMyc N-terminal fusion of beta-2 adrenergic receptor in mammalian cells, MMLV retroviral plasmid (MBIC5, dL5**, FAP, ADRB2)

Marcel Bruchez

Perkins et al Biochemistry. 2018 Feb 6;57(5):861-871. doi: 10.1021/acs.biochem.7b01135. Epub 2018 Jan 16.

https://media.addgene.org/snapgene-media/v2.0.0/sequences/235286/9e561f13-9553-448e-a852-f37fe0f95a96/addgene-plasmid-101253-sequence-235286.gbk

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LOCUS sequence-235286- 7794 bp DNA circular SYN 26-AUG-2024 DEFINITION Expresses HA-dL5(E52D)-cMyc N-terminal fusion of beta-2 adrenergic receptor in mammalian cells, MMLV retroviral plasmid (MBIC5, dL5**, FAP, ADRB2). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7794) AUTHORS Perkins LA, Yan Q, Schmidt BF, Kolodieznyi D, Saurabh S, Larsen MB, Watkins SC, Kremer L, Bruchez MP TITLE Genetically Targeted Ratiometric and Activated pH Indicator Complexes (TRApHIC) for Receptor Trafficking. JOURNAL Biochemistry. 2018 Feb 6;57(5):861-871. doi: 10.1021/acs.biochem.7b01135. Epub 2018 Jan 16. PUBMED 29283245 REFERENCE 2 (bases 1 to 7794) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1021/acs.biochem.7b01135"; journalName: "Biochemistry"; date: "2018-02-6- 6"; volume: "57"; issue: "5"; pages: "861-871" FEATURES Location/Qualifiers source 1..7794 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 66..535 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 600..799 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1000..1416 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1346..1368 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1384..1400 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" sig_peptide 1476..1538 /codon_start=1 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader /translation="METDTLLLWVLLLWVPGSTGD" CDS 1539..1565 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1581..2300 /codon_start=1 /product="fluorogen-activating protein composed of a dimer of the L5-MG single-chain antibody fragment (Szent-Gyorgyi et al., 2008)" /label=FAP-alpha-2 /note="binds malachite green derivatives to produce red fluorescence" /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL SDVDGYLFGGGTQLTVLS" CDS 2337..2366 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3618..3764 /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" promoter 3804..3834 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3842..3858 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 3847..3869 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3866..3882 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3866..3882 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 3878..4156 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWR" primer_bind complement(3898..3915) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3898..3914) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3907..3929) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(4185..4205) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4190..4516 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4370..4502 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4432..4451 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4526..5125 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4526..4545) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5022..5042 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 5346..5815 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(5945..5962) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6116..6704) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6196..6215) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6864..7724) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7487..7506 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(join(7725..7794,1..35)) /gene="bla" /label=AmpR promoter ORIGIN 1 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 61 gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg 121 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc 181 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt 241 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt 301 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc 361 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc 421 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg 481 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg 541 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca 601 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg 661 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact 721 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt 781 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg 841 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg 901 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt 961 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc 1021 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta 1081 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc 1141 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc 1201 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc 1261 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct 1321 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc 1381 tccctttatc cagccctcac tccttctcta ggcgccggcc ggatccacta gtaacggccg 1441 ccagtgtgct ggaattcggc ttggggatat ccaccatgga gacagacaca ctcctgctat 1501 gggtactgct gctctgggtt ccaggttcca ctggtgacta tccatatgat gttccagatt 1561 atgctggggc ccagccggcc caggccgtcg ttacccaaga acctagtgtt accgttagcc 1621 caggtggtac tgttatactt acttgtggaa gtggtacggg tgccgtcaca tctggtcatt 1681 atgcaaattg gtttcaacaa aaaccaggac aagctccaag agctttgatt tttgatactg 1741 ataagaagta ttcttggacc ccaggtagat tttctggatc tttgctggga gcaaaggcag 1801 ctttgacaat atcagatgct cagcctgagg acgaagccga gtattactgt tctcttagcg 1861 acgtggatgg ctacttgttt ggcggtggaa cacaactgac ggttctgtcc ggtggtggcg 1921 gctctggtgg cggtggcagc ggcggtggtg gttccggagg cggcggttct caggctgtgg 1981 tgactcagga gccgtcagtg actgtgtccc caggagggac agtcattctc acttgtggct 2041 ccggcactgg agctgtcacc agtggtcatt atgccaactg gttccagcag aagcctggcc 2101 aagcccccag ggcacttata tttgacaccg acaagaagta ttcctggacc cctggccgat 2161 tctcaggctc cctccttggg gccaaggctg ccctgaccat ctcggatgcg cagcctgaag 2221 atgaggctga gtattactgt tcgctctccg acgttgacgg ttatctgttc ggaggaggca 2281 cccagctgac cgtcctctcc ggccgcaggg gccgggatcc gcggctgcag gtcgacgaac 2341 aaaaactcat ctcagaagag gatctgaatg ctatggggca acccgggaac ggcagcgcct 2401 tcttgctggc acccaatgga agccatgcgc cggaccacga cgtcacgcag caaagggacg 2461 aggtgtgggt ggtgggcatg ggcatcgtca tgtctctcat cgtcctggcc atcgtgtttg 2521 gcaatgtgct ggtcatcaca gccattgcca agttcgagcg tctgcagacg gtcaccaact 2581 acttcatcac ttcactggcc tgtgctgatc tggtcatggg cctagcagtg gtgccctttg 2641 gggccgccca tattcttatg aaaatgtgga cttttggcaa cttctggtgc gagttttgga 2701 cttccattga tgtgctgtgc gtcacggcca gcattgagac cctgtgcgtg atcgcagtgg 2761 atcgctactt tgccattact tcacctttca agtaccagag cctgctgacc aagaataagg 2821 cccgggtgat cattctgatg gtgtggattg tgtcaggcct tacctccttc ttgcccattc 2881 agatgcactg gtacagggcc acccaccagg aagccatcaa ctgctatgcc aatgagacct 2941 gctgtgactt cttcacgaac caagcctatg tcattgcctc ttccatcgtg tccttctacg 3001 ttcccctggt gatcatggtc ttcgtctact ccagggtctt tcaggaggcc aaaaggcagc 3061 tccagaagat tgacaaatct gagggccgct tccatgtcca gaaccttagc caggtggagc 3121 aggatgggcg gacggggcat ggactccgca gatcttccaa gttctgcttg aaggagcaca 3181 aagccctcaa gacgttaggc atcatcatgg gcactttcac cctctgctgg ctgcccttct 3241 tcatcgttaa cattgtgcat gtgatccagg ataacctcat ccgtaaggaa gtttacatcc 3301 tcctaaattg gataggctat gtcaattctg gtttcaatcc ccttatctac tgccggagcc 3361 cagatttcag gattgccttc caggagcttc tgtgcctgcg caggtcttct ttgaaggcct 3421 atggcaatgg ctactccagc aacggcaaca caggggagca gagtggatat cacgtggaac 3481 aggagaaaga aaataaactg ctgtgtgaag acctcccagg cacggaagac tttgtgggcc 3541 atcaaggtac tgtgcctagc gataacattg attcacaagg gaggaattgt agtacaaatg 3601 actcactgct gtagaatgct gtgggccagg acacgcagga ggtcatcgtg gtgccacact 3661 ccttgccctt taaggtggtg gtgatctcag ccatcctggc cctggtggtg ctcaccatca 3721 tctcccttat catcctcatc atgctttggc agaagaagcc acgtccacag catgggttag 3781 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgaa 3841 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact 3901 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct 3961 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc 4021 ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga 4081 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc 4141 ctcaaactgg cgttagccag caggtggcgt tagctcgacc ctgtggaatg tgtgtcagtt 4201 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 4261 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 4321 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 4381 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 4441 agaggccgag gccgcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 4501 cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac 4561 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 4621 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 4681 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 4741 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 4801 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct 4861 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 4921 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga 4981 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 5041 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 5101 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag 5161 gagcgcacga ccccatgcat cgataaaata aaagatttta tttagtctcc agaaaaaggg 5221 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa 5281 ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg 5341 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg 5401 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc 5461 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt 5521 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt 5581 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc 5641 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc 5701 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg 5761 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacatgc 5821 agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt 5881 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 5941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 6001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 6061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 6121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 6181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 6241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 6301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 6361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 6421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 6481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 6541 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 6601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 6661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 6721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 6781 gattatcaaa aaggatcttc acctagatcc ttttgcggcc gcaaatcaat ctaaagtata 6841 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6901 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6961 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 7021 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 7081 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 7141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 7201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7621 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7681 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7741 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaa //