Name
stringlengths 1
127
| ID
stringlengths 1
6
| Flame
stringclasses 4
values | Purpose
stringlengths 0
474
| Depositing Lab
stringlengths 0
42
| Publication
stringlengths 0
220
| GenBank File
stringlengths 0
144
| Sequence Type
stringclasses 2
values | GenBank Raw
stringlengths 0
181k
| Backbone
dict | Growth in Bacteria
dict | Terms and Licenses
dict | References
dict | Gene/Insert 1
dict | Cloning Information for Gene/Insert 1
dict | Gene/Insert 2
dict | Cloning Information for Gene/Insert 2
dict | Gene/Insert 3
dict | Cloning Information for Gene/Insert 3
dict |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
pBACgus/LRRK2 1867-2176 C-His | 40378 | Michael J Fox Foundation MJFF | Michael J Fox Foundation plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/52055/33e78450-fb17-4c44-8791-65bf64b343dc/addgene-plasmid-40378-sequence-52055.gbk | full | LOCUS sequence-52055-e 8344 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 8344)
AUTHORS MJFF
TITLE Michael J Fox Foundation plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8344)
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"Unpublished"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..8344
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 329..1156
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
CDS 329..453
/codon_start=3
/product="baculovirus late expression factor 2"
/label=lef2
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS complement(491..1096)
/codon_start=1
/product="baculovirus ORF603 protein"
/label=ORF603
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
CDS complement(1167..2978)
/codon_start=1
/gene="uidA (or gusA) from E. coli"
/product="beta-glucuronidase"
/label=GUS
/translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
ADEHGIVVIDETAAVGFSLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
promoter complement(2987..3302)
/gene="Autographa californica p6.9"
/label=p6.9 promoter
/note="baculovirus late basic protein promoter"
promoter 3317..3408
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 3333..3350
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
CDS 4351..4368
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 4394..4411
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_recomb 4448..5805
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
CDS complement(4464..5805)
/codon_start=2
/product="baculovirus capsid-associated protein"
/label=ORF1629
/note="required for viral replication"
/translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
primer_bind complement(5897..5914)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6068..6656)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6148..6167)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6827..7687)
/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 7450..7469
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7688..7792)
/gene="bla"
/label=AmpR promoter
rep_origin 7819..8274
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7906..7925)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8116..8137
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 ggaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag
61 agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtcga catgctgaac
121 aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat
181 gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg
241 gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac
301 cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat
361 aaaccaccaa actgccaaaa aatgaaaact gtcgacaagc tctgtccgtt tgctggcaac
421 tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata aatgctaaat
481 ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa
541 ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac
601 agttaatttg cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt
661 cttcgtattc cttctctttt tcatttttct cttcataaaa attaacatag ttattatcgt
721 atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt
781 ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat
841 tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt
901 tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa
961 ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt
1021 taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt
1081 taaaaataac agccattgta atgagacgca caaactaata tcacaaactg gaaatgtcta
1141 tcaatatata gttgctgatc agatcctcat tgtttgcctc cctgctgcgg tttttcaccg
1201 aagttcatgc cagtccagcg tttttgcagc agaaaagccg ccgacttcgg tttgcggtcg
1261 cgagtgaaga tccctttctt gttaccgcca acgcgcaata tgccttgcga ggtcgcaaaa
1321 tcggcgaaat tccatacctg ttcaccgacg acggcgctga cgcgatcaaa gacgcggtga
1381 tacatatcca gccatgcaca ctgatactct tcactccaca tgtcggtgta cattgagtgc
1441 agcccggcta acgtatccac gccgtattcg gtgatgataa tcggctgatg cagtttctcc
1501 tgccaggcca gaagttcttt ttccagtacc ttctctgccg tttccaaatc gccgctttgg
1561 acataccatc cgtaataacg gttcaggcac agcacatcaa agagatcgct gatggtatcg
1621 gtgtgagcgt cgcagaacat tacattgacg caggtgatcg gacgcgtcgg gtcgagttta
1681 cgcgttgctt ccgccagtgg cgcgaaatat tcccgtgcac cttgcggacg ggtatccggt
1741 tcgttggcaa tactccacat caccacgctt gggtggtttt tgtcacgcgc tatcagctct
1801 ttaatcgcct gtaagtgcgc ttgctgagtt tccccgttga ctgcctcttc gctgtacagt
1861 tctttcggct tgttgcccgc ttcgaaacca atgcctaaag agagcgaaaa gccgacagca
1921 gcagtttcat caatcaccac gatgccatgt tcatctgccc agtcgagcat ctcttcagcg
1981 taagggtaat gcgaggtacg gtaggagttg gccccaatcc agtccattaa tgcgtggtcg
2041 tgcaccatca gcacgttatc gaatcctttg ccacgtaagt ccgcatcttc atgacgacca
2101 aagccagtaa agtagaacgg tttgtggtta atcaggaact gttggccctt cactgccact
2161 gaccggatgc cgacgcgaag cgggtagata tcacactctg tctggctttt ggctgtgacg
2221 cacagttcat agagataacc ttcacccggt tgccagaggt gcggattcac cacttgcaaa
2281 gtcccgctag tgccttgtcc agttgcaacc acctgttgat ccgcatcacg cagttcaacg
2341 ctgacatcac cattggccac cacctgccag tcaacagacg cgtggttaca gtcttgcgcg
2401 acatgcgtca ccacggtgat atcgtccacc caggtgttcg gcgtggtgta gagcattacg
2461 ctgcgatgga ttccggcata gttaaagaaa tcatggaagt aagactgctt tttcttgccg
2521 ttttcgtcgg taatcaccat tcccggcggg atagtctgcc agttcagttc gttgttcaca
2581 caaacggtga tacgtacact tttcccggca ataacatacg gcgtgacatc ggcttcaaat
2641 ggcgtatagc cgccctgatg ctccatcact tcctgattat tgacccacac tttgccgtaa
2701 tgagtgaccg catcgaaacg cagcacgata cgctggcctg cccaaccttt cggtataaag
2761 acttcgcgct gataccagac gttgcccgca taattacgaa tatctgcatc ggcgaactga
2821 tcgttaaaac tgcctggcac agcaattgcc cggctttctt gtaacgcgct ttcccaccaa
2881 cgctgatcaa ttccacagtt ttcgcgatcc agactgaatg cccacaggcc gtcgagtttt
2941 ttgatttcac gggttggggt ttctacagga cggaccatat ttataggtgt aatttatgta
3001 gctgtaattt ttaccttatt aatatttttt acgctttgca ttcgacgact gaactcccaa
3061 atatatgttt aactcgtctt ggtcgtttga atttttgttg ctgtgtttcc taatattttc
3121 catcacctta aatatgttat tgtaatcctc aatgttgaac ttgcaattgg acacggcata
3181 gttttccata gtcgtgtaaa acatggtatt ggctgcattg taatacatcc gactgagcgg
3241 gtacggatct atgtgtttga gcagcctgtt caaaaactct gcatcgtcgc aaaacggaat
3301 ttcgtcgaca gatctgatca tggagataat taaaatgata accatctcgc aaataaataa
3361 gtattttact gttttcgtaa cagttttgta ataaaaaaac ctataaatat aggatccatg
3421 aacatcatgc tcaacaacga tgagctcgaa tttgaacaag ctccagaatt cctgctcggc
3481 gacggctctt tcggctccgt ataccgtgct gcttatgaag gcgaagaagt cgccgtaaag
3541 atcttcaaca agcacacctc gctccgtctg ctccgtcagg aactggtcgt tctgtgtcat
3601 ctgcaccatc cgagcctcat ctccctgctg gccgctggca tccgtcctcg tatgctggtg
3661 atggagctgg catccaaggg ctctctcgac cgcctgctgc agcaggacaa ggcttctctg
3721 acccgtactc tgcagcatcg tatcgctctg catgtagctg acggcctgcg ttatcttcat
3781 tctgcaatga tcatttatcg cgacctgaaa cctcataatg tactgctgtt tactctgtac
3841 ccgaacgctg ccatcatcgc taagattgca gactatggta ttgcgcagta ctgctgccgc
3901 atgggcatta aaacaagcga aggtacgccg ggtttccgtg ctccagaagt ggctcgtggc
3961 aacgttattt acaaccagca agctgacgtt tactcgttcg gtctgctcct gtacgatatc
4021 ctgactaccg gcggccgtat cgtagaaggt ctgaagttcc ctaacgaatt cgacgaactg
4081 gaaatccagg gcaagctgcc agacccggtt aaagaatacg gttgcgctcc gtggccgatg
4141 gtagagaagc tcatcaaaca gtgccttaaa gaaaaccctc aggaacgtcc aacctctgct
4201 caagtgttcg acatccttaa cagcgctgaa ctcgtctgtc tgactcgccg tattctcctg
4261 cctaagaatg ttatcgtgga atgtatggtt gctactcacc ataactctcg caacgcttcc
4321 atctggctcg gctgtggcca cactgaccgt caccaccatc accatcattg ataagcttgc
4381 ggccgcactc gagcaccacc accaccacca ctaacctagg tagctgagcg catgcaagct
4441 gatccgggtt attagtacat ttattaagcg ctagattctg tgcgttgttg atttacagac
4501 aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg gtatgttaga
4561 gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa atcctcaata
4621 gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga accgatggct
4681 ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg tagcagcaat
4741 ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa
4801 atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg actcgacgta
4861 aacacgttaa atagagcttg gacatattta acatcgggcg tgttagcttt attaggccga
4921 ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga ttttgccata
4981 gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt tgtagttgag
5041 ctttttggaa ttatttctga ttgcgggcgt ttttgggcgg gtttcaatct aactgtgccc
5101 gattttaatt cagacaacac gttagaaagc gatggtgcag gcggtggtaa catttcagac
5161 ggcaaatcta ctaatggcgg cggtggtgga gctgatgata aatctaccat cggtggaggc
5221 gcaggcgggg ctggcggcgg aggcggaggc ggaggtggtg gcggtgatgc agacggcggt
5281 ttaggctcaa atgtctcttt aggcaacaca gtcggcacct caactattgt actggtttcg
5341 ggcgccgttt ttggtttgac cggtctgaga cgagtgcgat ttttttcgtt tctaatagct
5401 tccaacaatt gttgtctgtc gtctaaaggt gcagcgggtt gaggttccgt cggcattggt
5461 ggagcgggcg gcaattcaga catcgatggt ggtggtggtg gtggaggcgc tggaatgtta
5521 ggcacgggag aaggtggtgg cggcggtgcc gccggtataa tttgttctgg tttagtttgt
5581 tcgcgcacga ttgtgggcac cggcgcaggc gccgctggct gcacaacgga aggtcgtctg
5641 cttcgaggca gcgcttgggg tggtggcaat tcaatattat aattggaata caaatcgtaa
5701 aaatctgcta taagcattgt aatttcgcta tcgtttaccg tgccgatatt taacaaccgc
5761 tcaatgtaag caattgtatt gtaaagagat tgtctcaagc tcggatcgat cccgcacgcc
5821 gataacaagc cttttcattt ttactacagc attgtagtgg cgagacactt cgctgtcgtc
5881 gaggtttaaa cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
5941 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
6001 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
6061 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
6121 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
6181 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
6241 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
6301 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
6361 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
6421 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
6481 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
6541 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
6601 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
6661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
6721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
6781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
6841 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
6901 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
6961 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
7021 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
7081 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
7141 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
7201 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
7261 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
7321 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
7381 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
7441 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
7501 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
7561 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
7621 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
7681 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
7741 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
7801 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
7861 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
7921 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
7981 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
8041 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
8101 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
8161 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
8221 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcccatt
8281 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
8341 gcca
//
| {
"Vector backbone": "pBACgus",
"Backbone size w/o insert (bp)": "",
"Vector type": "Baculovirus",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Top10",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "LRRK2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "WT aa1867-2176",
"Entrez Gene": "LRRK2",
"Promoter": "polH",
"Tag / Fusion Protein": "6xHis (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (destroyed during cloning)",
"3′ cloning site": "HindIII (destroyed during cloning)",
"5′ sequencing primer": "DSP2466For:GAATTTCGTCGACAGATC",
"3′ sequencing primer": "DSP2466Rev:AATAACCCGGATCAGCTT"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
Ube2r1 | 31426 | Rachel Klevit | Christensen et al Nat Struct Mol Biol. 2007 Oct . 14(10):941-8. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/37917/caff4ec9-b5e8-4547-9bf4-13320610ed74/addgene-plasmid-31426-sequence-37917.gbk | partial | LOCUS sequence-37917-e 812 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 812)
AUTHORS Christensen DE, Brzovic PS, Klevit RE
TITLE E2-BRCA1 RING interactions dictate synthesis of mono- or specific
polyubiquitin chain linkages.
JOURNAL Nat Struct Mol Biol. 2007 Oct . 14(10):941-8.
PUBMED 17873885
REFERENCE 2 (bases 1 to 812)
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 Struct
Mol Biol. 2007 Oct . 14(10):941-8."
FEATURES Location/Qualifiers
source 1..812
/mol_type="genomic DNA"
/organism="unspecified"
CDS 35..61
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 41..67
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/note="This is a variant of the standard HA tag, which has
sequence YPYDVPDYA. Some anti-HA antibodies may not
recognize this variant."
/translation="YDVPDYASL"
protein_bind 98..122
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 cccaccaaac ccaaaaaaag agatctctat ggcttaccca tacgatgttc cagattacgc
61 tagcttgggt ggtcatatgg ccatggaggc cggatctaca agtttgtaca aaaaagcagg
121 ctccgcggcc gcccccttca ccatggctcg gccgctagtg cccagctcgc agaaggcgct
181 gctgctggag ctcaaggggc tgcaggaaga gccggtcgag ggattccgcg tgacactggt
241 ggacgagggc gatctataca actgggaggt ggccatcttc gggcccccca acacctacta
301 cgagggcggc tacttcaagg cgcgcctcaa gttccccatc gactacccat actctccacc
361 agcctttcgg ttcctgacca agatgtggca ccctaacatc tacgagacgg gggacgtgtg
421 tatctccatc ctccacccgc cggtggacga cccccagagc ggggagctgc cctcagagag
481 gtggaacccc acgcagaacg tcaggaccat tctcctgagt gtgatctccc tcctgaacga
541 gcccaacacc ttctcgcccg caaacgtgga cgcctccgtg atgtacagga agtggaaaga
601 gagcaagggg aaggatcggg agtacacaga catcatccgg aagcaggtcc tggggaccaa
661 ggtggacgcg gagcgtgacg gcgtgaaggt gcccaccacg ctggccgagt actgcgtgaa
721 gaccaaggcg ccggcgcccg acgagggctc agacctcttc tacgacgact actacgagga
781 cggcgaggtg gaggaggagg ccgacagctg ct
//
| {
"Vector backbone": "pACT2",
"Backbone size w/o insert (bp)": "8100",
"Vector type": "Yeast Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "E2-BRCA1 RING interactions dictate synthesis of mono- or specific polyubiquitin chain linkages",
"Publication": "Nat Struct Mol Biol. 2007 Oct . 14(10):941-8.",
"DOI": "10.1038/nsmb1295",
"Authors": "Christensen DE, Brzovic PS, Klevit RE",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17873885",
"PubMed ID": "PubMed 17873885"
} | {
"Gene/Insert name": "Ubiquitin Conjugating Enzyme",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "400",
"Mutation": "",
"Entrez Gene": "CDC34",
"Promoter": "",
"Tag / Fusion Protein": "HA (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GAL4 AD",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pcDNA3.1hisA cdc25c C330S | 10966 | Toren Finkel | Savitsky et al J Biol Chem. 2002 Jun 7. 277(23):20535-40. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/17935/16b81622-0fe8-4f84-95c8-3c9aee8944cc/addgene-plasmid-10966-sequence-17935.gbk | partial | LOCUS sequence-17935-e 950 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 950)
AUTHORS Savitsky PA, Finkel T
TITLE Redox regulation of Cdc25C.
JOURNAL J Biol Chem. 2002 Jun 7. 277(23):20535-40.
PUBMED 11925443
REFERENCE 2 (bases 1 to 950)
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. 2002 Jun 7. 277(23):20535-40."
FEATURES Location/Qualifiers
source 1..950
/mol_type="genomic DNA"
/organism="unspecified"
CDS complement(19..60)
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
ORIGIN
1 gtgatgatga ccggtacgcg tagaatcgag accgaggaga gggttaggga taggcttacc
61 ttcgaagggc cctctagatg ggctcatgtc cttcaccaga agggcaatct gctcccgcag
121 ctgccgctcc ccttcctgca ctttgctctg gcttcgacac ctcagcaact cagtcttgtg
181 gtcctgatga tgcatagggc agtagctctg tggttcacac agttccatat attctggaaa
241 gaagtctctg tagccgcctt taaggatata tagctctggg tagtacaatg caggatactg
301 gttcagagac ctgtcctctt cacgcagaca gcggcacatt cgggggcccc tctctgagga
361 gaattcacag tggaacacga tgattattct cttctgggtg tccaaaggga cgatgggctt
421 cttcagaaag aagttaaaca gttcttcctg actatataag tttaaggctc cctggatgtg
481 tcctcccaga tactcataag gatagcgtga atcaatgaca taaaacttct caatcagacc
541 ctggaacttc cccgacagta aggcagccac tgtttctggg ttgacatact tcagatcttg
601 gtgtttccct gacacggttg gcagcgcaca taccttggaa aaatcaccaa tcaggtgccc
661 ctggttagaa tcttcctcca gcatctgagt gatagtaatg tcacacagag agactgtctt
721 ctttaaacat aagcccttcc tgagctttcc ttggccagaa aaatactttt ttttaacttt
781 atctggtatt gtgttgtcct tgaatttttc cacctgcttc agtcttggcc tgttcaagtt
841 ctctggcatc gacggggagc gatataggcc acttctgctc acctttgctt cttgatcttt
901 cagggaaaac tccattaatt catctgaaat ctcttctgcc tggtcttctc
//
| {
"Vector backbone": "pcDNA3.1 His A",
"Backbone size w/o insert (bp)": "5500",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Redox regulation of Cdc25C",
"Publication": "J Biol Chem. 2002 Jun 7. 277(23):20535-40.",
"DOI": "10.1074/jbc.M201589200",
"Authors": "Savitsky PA, Finkel T",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/11925443",
"PubMed ID": "PubMed 11925443"
} | {
"Gene/Insert name": "cdc25c C330S",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1450",
"Mutation": "C330S. Cysteine to Serine substitution at aa330.",
"Entrez Gene": "CDC25C",
"Promoter": "",
"Tag / Fusion Protein": "6xHis (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "BGHrev"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pHR_Gal4UAS_4D5-Low_CAR-mCherry_pGK_4D5-Low_SynNotch_RHL183 | 164827 | Lentiviral vector for Gal4 inducible AntiHER2 4D5-Low CAR-mCherry and constitutive expression of antiHER2 4D5-Low Gal4VP64 Synthetic Notch | Wendell Lim | Hernandez-Lopez et al Science. 2021 Mar 12;371(6534):1166-1171. doi: 10.1126/science.abc1855. Epub 2021 Feb 25. | partial | {
"Vector backbone": "pHR'SIN",
"Backbone size w/o insert (bp)": "",
"Vector type": "Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "T cell circuits that sense antigen density with an ultrasensitive threshold",
"Publication": "Science. 2021 Mar 12;371(6534):1166-1171. doi: 10.1126/science.abc1855. Epub 2021 Feb 25.",
"DOI": "10.1126/science.abc1855.",
"Authors": "Hernandez-Lopez RA, Yu W, Cabral KA, Creasey OA, Lopez Pazmino MDP, Tonai Y, De Guzman A, Makela A, Saksela K, Gartner ZJ, Lim WA /scienceabc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33632893",
"PubMed ID": "PubMed 33632893"
} | {
"Gene/Insert name": "AntiHER2 4D5-Low Chimeric Antigen receptor (CD8alphaTM-41BB-CD3z)",
"Species": "Synthetic",
"Insert Size (bp)": "1542",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "miniCMV",
"Tag / Fusion Protein": "mCherry (C terminal on insert)"
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "agagctcgtttagtgaaccg",
"3′ sequencing primer": "CAGCGGGGCTGCTAAAGCGCATGC"
} | {
"Gene/Insert name": "AntiHER2 4D5-Low Gal4VP64 synthetic Notch receptor",
"Species": "Synthetic",
"Insert Size (bp)": "2445",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "pGK",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GCTTCAAAAGCGCACGTCT",
"3′ sequencing primer": "catagcgtaaaaggagcaaca"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|||
PL2x2_5 | 45491 | David Baker | Mutations_Koga_Nature (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/64277/187cc730-3ffb-48fa-8c39-389b2c1a6991/addgene-plasmid-45491-sequence-64277.gbk | partial | LOCUS sequence-64277-e 855 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 855)
TITLE Mutations_Koga_Nature
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 855)
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..855
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 68..87
/label=T7
/note="T7 promoter, forward primer"
promoter 68..86
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 87..111
/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)."
RBS 126..148
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 474..491
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(544..562)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 558..605
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(729..748)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatcga tctcgatccc
61 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
121 ataattttgt ttaactttaa gaaggagata tacatatggg caaagtcctg ctggtgattt
181 ccaccgatac gaacattatc agctctgtcc aggaacgtgc gaaacataat tatccgggtc
241 gttacattcg caccgccacg agttcccaag atatccgcga cattatcaaa agcatgaaag
301 ataacggcaa accgctggtt gtgtttgtga atggtgcatc tcagaacgac gttaacgaat
361 ttcaaaacga agctaaaaaa gaaggcgtta gctatgatgt cctgaaatcg accgacccgg
421 aagaactgac gcagcgtgtt cgcgaatttc tgaaaaccgc cggtagcctc gagcaccacc
481 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
541 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
601 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
841 ccccaaaaac tgata
//
| {
"Vector backbone": "pET29b(+)",
"Backbone size w/o insert (bp)": "5370",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "XL10 Gold",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "PL2x2_5",
"Species": "Synthetic",
"Insert Size (bp)": "321",
"Mutation": "changed E30Y of Di-IV_5_PL2x2_6",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "6xHis (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pBT29 | 138521 | phage cloning | David Liu | Richter et al Nat Biotechnol. 2020 Jul;38(7):883-891. doi: 10.1038/s41587-020-0453-z. Epub 2020 Mar 16. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/273893/5422d40e-9421-4a4e-b7fe-f74178362b09/addgene-plasmid-138521-sequence-273893.gbk | full | LOCUS sequence-273893- 5662 bp DNA circular SYN 26-AUG-2024
DEFINITION phage cloning.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5662)
AUTHORS Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW,
Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR
TITLE Phage-assisted evolution of an adenine base editor with improved Cas
domain compatibility and activity.
JOURNAL Nat Biotechnol. 2020 Jul;38(7):883-891. doi:
10.1038/s41587-020-0453-z. Epub 2020 Mar 16.
PUBMED 32433547
REFERENCE 2 (bases 1 to 5662)
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/s41587-020-0453-z"; journalName: "Nat Biotechnol"; date:
"2020-07"; volume: "38"; issue: "7"; pages: "883-891"
FEATURES Location/Qualifiers
source 1..5662
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 23..1255
/codon_start=1
/gene="II"
/product="pII"
/label=M13 gene II
/note="replication"
/translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
TVEYHVDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
FGTTDLGLCSEALLLNFANSLPCLYDLLDVNATTISRIDAAFSARAPNENIAKQVIDHL
RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
DLWSFSFSELFKAFEGNSMNIYDDSAVLAAIQSKHFTITPSGKTSFAKASRYFGFYRRL
VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
DWYNEPVLKIA"
CDS 1267..1530
/codon_start=1
/gene="V"
/product="pV"
/label=M13 gene V
/note="replication"
/translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQFCYVDLGNEYPVLV
KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
CDS 1532..1633
/codon_start=1
/gene="VII"
/product="pVII"
/label=M13 gene VII
/note="minor coat protein"
/translation="MEQVADFDIIYQAMIQISVVLCFALGIIAGGQR"
CDS 1630..1728
/codon_start=1
/gene="IX"
/product="pIX"
/label=M13 gene IX
/note="minor coat protein"
/translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
CDS 1725..1946
/codon_start=1
/gene="VIII"
/product="pVIII"
/label=M13 gene VIII
/note="major coat protein"
/translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
GYAWAMVVVIVGATIGIKLFKKFTSKAS"
terminator 1953..1992
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
protein_bind 2003..2021
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2028..2046
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 2064..2075
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2082..2759
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
terminator 2777..2848
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2864..2891
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(2990..3805)
/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(2990..3805)
/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 3713..3732
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3797..3816
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS complement(3873..4823)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 4871..5093
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
ORIGIN
1 cttattatca accggggtac atatgattga catgctagtt ttacggttac cgttcatcga
61 ttctcttgtt tgctccagac tctcaggcaa tgacctgata gcctttgtag atctctcaaa
121 aatagctacc ctctccggca tgaatttatc agctagaacg gttgaatatc atgttgatgg
181 tgatttgact gtctccggcc tttcccaccc ttttgaatct ttacctacac attactcagg
241 cattgcattt aaaatatatg agggttctaa aaatttttat ccttgcgttg aaataaaggc
301 ttctcccgca aaagtattac aggggcataa tgtttttggt acaaccgatt taggtttatg
361 ctctgaggct ttattgctta attttgctaa ttctttgcct tgcctgtatg atttattgga
421 tgttaacgct actactatta gtagaattga tgccgccttt tcagcccgcg ccccaaatga
481 aaatatagct aaacaggtta ttgaccattt gcgaaatgta tctaatggtc aaactaaatc
541 tactcgttcg cagaattggg aatcaactgt tacatggaat gaaacttcca gacaccgtac
601 tttagttgca tatttaaaac atgttgagct acagcaccag attcagcaat taagctctaa
661 gccatccgca aaaatgacct cttatcaaaa ggaacaatta aaggtactct ctaatcctga
721 cctgttggag tttgcttccg gtctggttcg ctttgaggct cgaattaaaa cgagatattt
781 gaagtctttc gggcttcctc ttaatctttt tgatgcaatc cgctttgctt ctgactataa
841 tagtcagggt aaagacctga tttttgattt atggtcattc tcgttttctg aactgtttaa
901 agcatttgag gggaattcaa tgaatattta tgacgattcc gcagtattgg ccgctatcca
961 gtctaaacat tttactatta ccccctctgg caaaacttct tttgcaaaag cctctcgcta
1021 ttttggtttt tatcgtcgtc tggtaaacga gggttatgat agtgttgctc ttactatgcc
1081 tcgtaattcc ttttggcgtt atgtatctgc attagttgaa tgtggtattc ctaaatctca
1141 actgatgaat ctttctacct gtaataatgt tgttccgtta gttcgtttta ttaacgtaga
1201 tttttcttcc caacgtcctg actggtataa tgagccagtt cttaaaatcg cataaggtaa
1261 ttcacaatga ttaaagttga aattaaacca tctcaagccc aatttactac tcgttctggt
1321 gtttctcgtc agggcaagcc ttattcactg aatgagcagt tttgttacgt tgatttgggt
1381 aatgaatatc cggttcttgt caagattact cttgatgaag gtcagccagc ctatgcgccg
1441 ggtctgtaca ccgttcatct gtcctctttc aaagttggtc agttcggttc ccttatgatt
1501 gaccgtctgc gcctcgttcc ggctaagtaa catggagcag gtcgcggatt tcgacataat
1561 ttatcaggcg atgatacaaa tctccgttgt actctgtttc gcgcttggta taatagctgg
1621 gggtcaaaga tgagtgtttt agtgtattct ttcgcctctt tcgttttagg ttggtgcctc
1681 cgtagtggca ttacgtattt tacccgttta atggaaactt cctcatgaaa aagtctttag
1741 tcctcaaagc ctctgtagcc gttgctaccc tcgttccgat gctgtctttc gctgctgagg
1801 gtgacgatcc cgcaaaagcg gcctttgact ccctgcaagc ctcagcgacc gaatatatcg
1861 gttatgcgtg ggcgatggtt gttgtcattg tcggcgcaac tatcggtatc aagctgttta
1921 agaaattcac ctcgaaagca agttgataaa ctgatacaat taaaggctcc ttttggagcc
1981 tttttttttg gagtagaaga gctccctatc agtgatagag attgacatcc ctatcagtga
2041 tagagatact gagcacgaat ctgaaagagg agaaaggatc tatggcgagt agcgaagacg
2101 ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt
2161 tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga
2221 aagttaccaa aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt
2281 acggttccaa agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct
2341 tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg
2401 ttacccagga ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta
2461 ccaacttccc gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca
2521 ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac
2581 tgaaagacgg tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg
2641 ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag
2701 actacaccat cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat
2761 aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
2821 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
2881 gcctttctgc gtttatacag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
2941 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
3001 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
3061 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
3121 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
3181 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
3241 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
3301 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
3361 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
3421 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
3481 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
3541 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
3601 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
3661 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
3721 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
3781 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
3841 gttttattgt tcatgaaggc caaataggcc gttcagatcc ttccgtattt agccagtatg
3901 ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatgc
3961 ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt
4021 aaagcatcgt gtagtgtttt tcttagtccg ttacgtaggt aggaatctga tgtaatggtt
4081 gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc
4141 atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca
4201 accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat
4261 tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca
4321 tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac
4381 cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat
4441 tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct
4501 aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc
4561 aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca
4621 ccataagcat tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat
4681 accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat
4741 aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct
4801 ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac
4861 caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc
4921 tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat
4981 tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata
5041 aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt
5101 tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac
5161 agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat
5221 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
5281 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
5341 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
5401 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
5461 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
5521 atgcacccag taaggcagcg gtatcatcaa caggcttacc cgtcttactg tcaagaggac
5581 atccggtcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttgctgagga
5641 gacttaggga ccctgctctt ct
//
| {
"Vector backbone": "pCMV with a BR322 origin",
"Backbone size w/o insert (bp)": "",
"Vector type": "Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity",
"Publication": "Nat Biotechnol. 2020 Jul;38(7):883-891. doi: 10.1038/s41587-020-0453-z. Epub 2020 Mar 16.",
"DOI": "10.1038/s41587-020-0453-z.",
"Authors": "Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR /s---z",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32433547",
"PubMed ID": "PubMed 32433547"
} | {
"Gene/Insert name": "phage genes",
"Species": "Other",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
EGFP-uTEV1-MAVS-mTA6 | 171004 | HiLITR protease with MAVS [Y536delinsFIVLI, R537A, R538A] C-terminal tmd targeting information (Mitochondria) | Alice Ting | Coukos et al Elife. 2021 Aug 20;10. pii: 69142. doi: 10.7554/eLife.69142. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/335832/5ebb6d5d-f762-48db-9863-1e867104dc79/addgene-plasmid-171004-sequence-335832.gbk | full | LOCUS sequence-335832- 8608 bp DNA circular SYN 26-AUG-2024
DEFINITION HiLITR protease with MAVS [Y536delinsFIVLI, R537A, R538A] C-terminal
tmd targeting information (Mitochondria).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8608)
AUTHORS Coukos RW, Yao D, Sanchez ML, Strand ET, Olive ME, Udeshi ND,
Weissman JS, Carr SA, Bassik MC, Ting AY
TITLE An engineered transcriptional reporter of protein localization
identifies regulators of mitochondrial and ER membrane protein
trafficking in high-throughput CRISPRi screens.
JOURNAL Elife. 2021 Aug 20;10. pii: 69142. doi: 10.7554/eLife.69142.
PUBMED 34414886
REFERENCE 2 (bases 1 to 8608)
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: "2021-08-20"; volume: "10. pii: 69142. doi"; pages: "
10.7554/eLife.69142"
FEATURES Location/Qualifiers
source 1..8608
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 177..196
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 436..457
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 499..508
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(525..839)
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
primer_bind complement(528..552)
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 599..617
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 635..653
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 670..688
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 706..724
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 742..760
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 777..795
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 813..831
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 893..1010
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(1223..1267)
/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 1452..1685
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2178..2303
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 2350..2530
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(2531..2757)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
primer_bind complement(2785..2805)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2785..2803)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2824..2840)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2824..2840)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2837..2859)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2848..2864
/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(2872..2902)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2917..2938
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3055..3072)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3226..3814)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3306..3325)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3985..4845)
/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 4608..4627
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4846..4917)
/gene="bla"
/label=AmpR promoter
rep_origin 4917..5052
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 5092..5213
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5103..5122)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5157..5176
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 5285..5518
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS 5723..5734
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 6132..6152
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6192..6938)
/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"
promoter complement(6948..7458)
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind complement(7011..7029)
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS complement(join(8394..8608,1..502))
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS complement(join(8394..8608,1..502))
/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(8436..8457)
/label=EGFP-C
/note="EGFP, forward primer"
ORIGIN
1 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
61 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
121 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
181 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
241 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
301 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
361 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
421 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
481 gctcctcgcc cttgctcacc atggtggcgc tagccaattc tccaggcgat ctgacggttc
541 actaaacgag ctctgcttat ataggcctcc caccgtacac gcctacctcg acatacgttc
601 tctatcactg atagggagta aactcgacat acgttctcta tcactgatag ggataaactc
661 gacatacgtt ctctatcact gatagggagt aaactcgaca tacgttctct atcactgata
721 gggagtaaac tcgacatacg ttctctatca ctgataggga gtaaactcga catcgttctc
781 tatcactgat agggagtaaa ctcgacatac gttctctatc actgataggg agtaaactcg
841 acatatcgat tcgcggccaa agtggatctc tgctgtccct gtaataaacc cgaaaatttt
901 gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta
961 ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga
1021 atactgccat ttgtctcgag gtcgagaatt gtcccctcgg ggttgggagg tgggtctgaa
1081 acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta
1141 ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat
1201 ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt
1261 cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat
1321 tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt
1381 tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac
1441 cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct
1501 ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc
1561 tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca
1621 gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag
1681 ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg
1741 ctactcctaa tggttcaatt tttactactt tatatttata taattcactt ctccaattgt
1801 ccctcatatc tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta
1861 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
1921 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
1981 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
2041 aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
2101 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
2161 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
2221 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
2281 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
2341 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
2401 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
2461 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
2521 agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg caatgcggca
2581 attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat aaggcacgat
2641 cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctctcct tgtaaggcat
2701 gttgctaact catcgttacc atgttgcaag actacaagag tattgcataa gactacatta
2761 agcttgcagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
2821 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
2881 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
2941 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
3001 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
3061 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
3121 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
3181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
3241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
3301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
3361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
3421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
3481 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
3541 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
3601 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
3661 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
3721 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
3781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
3841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
3901 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
3961 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
4021 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
4081 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
4141 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
4201 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
4261 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
4321 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
4381 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
4441 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
4501 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
4561 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
4621 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
4681 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
4741 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
4801 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
4861 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaaaggcc
4921 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
4981 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
5041 taggggcggg atgggcggag ttaggggcgg gatagctaga gccagacatg ataagataca
5101 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
5161 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttcctctca
5221 ctctctgata ttcatttctt tgcaagttat aaatactgaa taataagatg acatgaacta
5281 ctactgctag agattttcca cactgactaa aagggtctga gggatctcta gttaccagag
5341 tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta
5401 agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga
5461 gacccagtac aagcaaaaag cagatcttgt cttcgttggg agtgaattag cccttccagt
5521 cccccctttt cttttaaaaa gtggctaaga tctacagctg ccttgtaagt cattggtctt
5581 aaaggtacct ctagtccgga cgccgaggcg aaacaggcgg ggaggcggcc caaagggaga
5641 tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc
5701 cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc
5761 gcagaatcca ggtggcaaca caggcgagca gccatggaaa ggacgtcagc ttccccgaca
5821 acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
5881 cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct
5941 gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca
6001 cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat
6061 acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag
6121 cagcgtatcc acatagcgta aaaggagcaa catagttaag aatacgatat cttggatcga
6181 tccttactta gttacccggg gagcatgtca aggtcaaaat cgtcaagagc gtcagcaggc
6241 agcatatcaa ggtcaaagtc gtcaagggca tcggctggga gcatgtctaa gtcaaaatcg
6301 tcaagggcgt cggtcggccc gccgctttcg cactttagct gtttctccag gccacatatg
6361 attagttcca ggccgaaaag gaaggcaggt tcggctccct gccggtcgaa cagctcaatt
6421 gcttgtctca gaagtggggg catagaatcg gtggtaggtg tctctctttc ctcttttgct
6481 acttgatgct cctgttcctc caatacgcag cccagtgtaa agtggcccac ggcggacaga
6541 gcgtacagtg cgttctccag ggagaagcct tgctgacaca ggaacgcgag ctgattttcc
6601 agggtttcgt actgtttctc tgttgggcgg gtgccgagat gcactttagc cccgtcgcga
6661 tgtgagagga gagcacagcg gtatgacttg gcgttgttcc gcagaaagtc ttgccatgac
6721 tcgccttcca gggggcagaa gtgggtatga tgcctgtcca gcatctcgat tggcagggca
6781 tcgagcaggg cccgcttgtt cttcacgtgc cagtacaggg taggctgctc aactcccagc
6841 ttttgagcga gtttccttgt cgtcaggcct tcgataccga caccattgag taattccaga
6901 gctccgttta tgactttgct cttgtccagt ctagacatgg tgaattgctg gggagagagg
6961 tcggtgattc ggtcaacgag ggagccgact gccgacgtgc gctccggagg cttgcagaat
7021 gcggaacacc gcgcgggcag gaacagggcc cacactaccg ccccacaccc cgcctcccgc
7081 accgcccctt cccggccgct gctctcggcg cgccctgctg agcagccgct attggccaca
7141 gcccatcgcg gtcggcgcgc tgccattgct ccctggcgct gtccgtctgc gagggtacta
7201 gtgagacgtg cggcttccgt ttgtcacgtc cggcacgccg cgaaccgcaa ggaaccttcc
7261 cgacttaggg gcggagcagg aagcgtcgcc ggggggccca caagggtagc ggcgaagatc
7321 cgggtgacgc tgcgaacgga cgtgaagaat gtgcgagacc cagggtcggc gccgctgcgt
7381 ttcccggaac cacgcccaga gcagccgcgt ccctgcgcaa acccagggct gccttggaaa
7441 aggcgcaacc ccaaccccgg atgaattcgg ccctagtgca gagccgcccg gatcagtacg
7501 atgaacagca ccaccaggag tgtgactacc agcacccctg tcacagccac ctggagccac
7561 agagccccag gtgagggcct tccactacct gagccggaac tgccgctccc ctctccactc
7621 ccaggcttac cagacccaga agtagagccg ctaccctcgc ctgatccggg cttgccagaa
7681 cctgacgtgg aaccgaattc caccatgaaa actttatggc ccccccacaa tactgagtca
7741 gcatttaatc gccaaccact aacccactgc tgcgcctcct gatttgtcaa caattccatg
7801 aagtttttcg gcacgcttgt gaaataattg tttgtgttgg tgaaattcga tgctgagtgt
7861 ataccaacaa tgaacccatc tctagttgat actaatggat tgccacactg cccatccttg
7921 gtttgaatcc aatgcttcca gaatatgcca tcagatgaag ggaatgtgca actagtgtct
7981 gacaccatgc tagacatgct cttagtttgg aagttggttg tcacaagaca tatgcgctct
8041 tccctttgtg gctctctaaa tttcagcttt tgaggaaatg gtgggaaatc cttaggcatg
8101 cgaataatta tcatgtccct cccatcaatg aggtgttgtt gcaaagtcgt ggtgttcttg
8161 accttgaata caccatgtag tgattggacc aacagtgttc cattatttct tctaaacaag
8221 tgcttgtttg taatgatgaa gggaccaaat ccaataccat acaacgatgt tgtgtgccca
8281 tcagattcat tcgtcaaatg acaaatggtg ctcgatatcg ggttgtaatc acgtggtccc
8341 ttaaacaagc tttctccgcc tgaaccggat ccgcctgtgc cagatccaag gtccttgtac
8401 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
8461 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
8521 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
8581 acgctgccgt cctcgatgtt gtggcgga
//
| {
"Vector backbone": "pCW-Cas9",
"Backbone size w/o insert (bp)": "8000",
"Vector type": "Mammalian Expression, Lentiviral, Synthetic Biology",
"Selectable markers": "EGFP"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "An engineered transcriptional reporter of protein localization identifies regulators of mitochondrial and ER membrane protein trafficking in high-throughput CRISPRi screens",
"Publication": "Elife. 2021 Aug 20;10. pii: 69142. doi: 10.7554/eLife.69142.",
"DOI": "10.7554/eLife.69142.",
"Authors": "Coukos RW, Yao D, Sanchez ML, Strand ET, Olive ME, Udeshi ND, Weissman JS, Carr SA, Bassik MC, Ting AY /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34414886",
"PubMed ID": "PubMed 34414886"
} | {
"Gene/Insert name": "EGFP-uTEV1-MAVS(tmd)[Y536delinsFIVLI, R537A, R538A]",
"Species": "Synthetic",
"Insert Size (bp)": "1600",
"Mutation": "MAVS(tmd)[Y536delinsFIVLI, R537A, R538A]",
"Entrez Gene": "",
"Promoter": "pTRE-tight (TetOn)",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pABY-T1 | 201764 | For C-terminal tagging of a protein-of-interest with the ALFA tag using S2/S3 primers in yeast | Saravanan Palani | Akhuli et al mSphere. 2022 Oct 26;7(5):e0033322. doi: 10.1128/msphere.00333-22. Epub 2022 Oct 3. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/403474/79d6cb09-9c6f-48cd-a477-6d6c8924c7cc/addgene-plasmid-201764-sequence-403474.gbk | full | LOCUS sequence-403474- 3956 bp DNA circular SYN 26-AUG-2024
DEFINITION For C-terminal tagging of a protein-of-interest with the ALFA tag
using S2/S3 primers in yeast.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3956)
AUTHORS Akhuli D, Dhar A, Viji AS, Bhojappa B, Palani S
TITLE ALIBY: ALFA Nanobody-Based Toolkit for Imaging and Biochemistry in
Yeast.
JOURNAL mSphere. 2022 Oct 26;7(5):e0033322. doi: 10.1128/msphere.00333-22.
Epub 2022 Oct 3.
PUBMED 36190134
REFERENCE 2 (bases 1 to 3956)
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: "mSphere.";
date: "2022-10-26"; volume: "7(5):e0033322. doi"; pages: "
10.1128/msphere.00333-22. Epub 2022 Oct 3"
FEATURES Location/Qualifiers
source 1..3956
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(31..50)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(32..50)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(137..154)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(308..896)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(388..407)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1067..1927)
/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 1690..1709
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1928..2032)
/gene="bla"
/label=AmpR promoter
primer_bind 2100..2118
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2156..2178)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2278..2297
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 2378..2396
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2378..2395
/label=SP6
/note="SP6 promoter, forward primer"
CDS 2440..2478
/codon_start=1
/product="rationally designed epitope tag that forms a
stable alpha-helix (Gotzke et al., 2019)"
/label=ALFA-tag
/note="It may be beneficial to flank the ALFA-tag sequence
with proline residues to preserve the helical structure."
/translation="SRLEEELRRRLTE"
gene 2527..3883
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 2527..2870
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 2871..3680
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(2938..2957)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 3686..3883
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
ORIGIN
1 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
61 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
121 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
721 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
841 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
901 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
961 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
1021 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
1081 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
1141 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
1201 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
1261 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
1321 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
1381 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
1441 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
1501 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
1561 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
1621 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
1681 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
1741 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
1801 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
1861 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
1921 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
1981 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
2041 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
2101 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
2161 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
2221 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
2281 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct
2341 acaattaata cataacctta tgtatcatac acatacgatt taggtgacac tatagaacgc
2401 ggccgccagc tgaagcttcg tacgctgcag gtcgacccat caagattaga agaggagctg
2461 aggagaagat tgaccgagta aagatctgtt tagcttgcct cgtccccgcc gggtcacccg
2521 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
2581 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
2641 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
2701 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
2761 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
2821 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg
2881 aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
2941 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
3001 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
3061 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
3121 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag
3181 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
3241 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
3301 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
3361 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt
3421 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
3481 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
3541 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
3601 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
3661 tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt
3721 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
3781 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
3841 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
3901 acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactag
//
| {
"Vector backbone": "pFA6a",
"Backbone size w/o insert (bp)": "3917",
"Vector type": "Yeast Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "ALIBY: ALFA Nanobody-Based Toolkit for Imaging and Biochemistry in Yeast",
"Publication": "mSphere. 2022 Oct 26;7(5):e0033322. doi: 10.1128/msphere.00333-22. Epub 2022 Oct 3.",
"DOI": "10.1128/msphere.00333-22.",
"Authors": "Akhuli D, Dhar A, Viji AS, Bhojappa B, Palani S /msphere-",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36190134",
"PubMed ID": "PubMed 36190134"
} | {
"Gene/Insert name": "ALFA tag",
"Species": "Synthetic",
"Insert Size (bp)": "39",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SalI (not destroyed)",
"3′ cloning site": "BglII (not destroyed)",
"5′ sequencing primer": "SP6",
"3′ sequencing primer": "Kan-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pFTK077 | 171349 | Part Plasmid Entry Vector (LVL0) from the Fungal Modular Cloning ToolKit. | Arnold Driessen | Mozsik et al ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/365609/7ba90bb3-503f-4d36-9af5-05016fc2ba67/addgene-plasmid-171349-sequence-365609.gbk | full | LOCUS sequence-365609- 2753 bp DNA circular SYN 26-AUG-2024
DEFINITION Part Plasmid Entry Vector (LVL0) from the Fungal Modular Cloning
ToolKit..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2753)
AUTHORS Mozsik L, Pohl C, Meyer V, Bovenberg RAL, Nygard Y, Driessen AJM
TITLE Modular Synthetic Biology Toolkit for Filamentous Fungi.
JOURNAL ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260.
PUBMED 34726388
REFERENCE 2 (bases 1 to 2753)
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. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260."
FEATURES Location/Qualifiers
source 1..2753
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(24..42)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 110..214
/gene="bla"
/label=AmpR promoter
CDS 436..1227
/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 1398..1986
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1887..1906
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2140..2157
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
61 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
181 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
301 tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
361 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
421 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
481 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
541 tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt tacggtgacc
601 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
661 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
721 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
781 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
841 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
901 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
961 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
1021 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
1081 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
1141 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta cgtaaaaggc
1201 gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata tatactttag
1261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
1321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
1381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
1441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
1501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
1561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
1621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
1681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
1741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
1801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
1861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
1921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
1981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
2041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
2101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
2161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatca
2221 ctctgtggtc tcagcttctt caaccgactc cggcttcact tcttcgcccg cactgcctac
2281 cgtttgtacc atctgactca tataaatgtc tagcccctac ctacactata cctaagggag
2341 agaagcgtag agtgattaac gtacgggcct atagtacccc gatctctaga tagaacattt
2401 agtagagatt aggatgccta actaatttaa cttgagcatt gtcccgttca tattgatttt
2461 cagtccatta tacactctta atcgtttccc ggtagaagcc tgatatatac gaccataggg
2521 tgtggagaac agggcttccc gtctgcttgg ccgtacttaa gctatatatt ctacacggcc
2581 aatactcaat gtgcccttag cacctaagcg gcactctagg gtaagtgcgg gtgatatagg
2641 tgagaagtct taagactgat gacaggatat cacgcgttac cctgcaccgt acctactacc
2701 ttcaatatca actctttcag gatggacagg gtcgacacgc ttgagaccac gaa
//
| {
"Vector backbone": "pICH41276",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression, Yeast Expression, Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Modular Synthetic Biology Toolkit for Filamentous Fungi",
"Publication": "ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260.",
"DOI": "10.1021/acssynbio.1c00260.",
"Authors": "Mozsik L, Pohl C, Meyer V, Bovenberg RAL, Nygard Y, Driessen AJM /acssynbioc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34726388",
"PubMed ID": "PubMed 34726388"
} | {
"Gene/Insert name": "TpenDE Pc21g21370 terminator",
"Species": "Penicillium, Aspergillus, or other fungal sources",
"Insert Size (bp)": "",
"Mutation": "n/a",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Unknown (unknown if destroyed)",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pM9-BetaGal-8C | 172489 | Plasmid encoding E. coli BetaGalactosidase with an N-terminal M9 nuclear localization sequence | Siegfried Musser | Chowdhury et al Nat Cell Biol. 2022 Jan;24(1):112-122. doi: 10.1038/s41556-021-00815-6. Epub 2022 Jan 10. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/356067/6e117671-b940-434d-80f4-3f8025689df6/addgene-plasmid-172489-sequence-356067.gbk | full | LOCUS sequence-356067- 8613 bp DNA circular SYN 26-AUG-2024
DEFINITION Plasmid encoding E. coli BetaGalactosidase with an N-terminal M9
nuclear localization sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8613)
AUTHORS Chowdhury R, Sau A, Musser SM
TITLE Super-resolved 3D tracking of cargo transport through nuclear pore
complexes.
JOURNAL Nat Cell Biol. 2022 Jan;24(1):112-122. doi:
10.1038/s41556-021-00815-6. Epub 2022 Jan 10.
PUBMED 35013558
REFERENCE 2 (bases 1 to 8613)
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/s41556-021-00815-6"; journalName: "Nat Cell Biol"; date:
"2022-01"; volume: "24"; issue: "1"; pages: "112-122"
FEATURES Location/Qualifiers
source 1..8613
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 48..503
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(135..154)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 345..366
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(595..1410)
/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(595..1410)
/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 1318..1337
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1402..1421
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 1532..2120
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2021..2040
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2274..2291
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2306..2445
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2371..2390)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2531..2553
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2547..2738)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3547..4629)
/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 4591..4610
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4630..4707)
/gene="lacI"
/label=lacI promoter
primer_bind 4913..4932
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5020..5039
/label=T7
/note="T7 promoter, forward primer"
promoter 5020..5038
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5039..5063
/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)."
RBS 5078..5100
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5111..5128
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5138..5155
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 5162..5206
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
CDS 5222..5236
/codon_start=1
/product="enterokinase recognition and cleavage site"
/label=enterokinase site
/translation="DDDDK"
CDS 5284..5295
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
CDS 8493..8510
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8563..8581)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator join(8577..8613,1..11)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag
61 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
121 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
181 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
241 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
301 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
361 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
421 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
481 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
541 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
601 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
661 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
721 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
781 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
841 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
901 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
961 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
1021 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
1081 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
1141 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
1201 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
1261 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
1321 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
1381 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
1441 agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
1501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
1561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
1621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
1681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
1741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
1801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
1861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
1921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
1981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
2041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
2101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
2161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
2221 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
2281 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
2341 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
2401 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
2461 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
2521 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
2581 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
2641 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
2701 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
2761 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
2821 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
2881 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
2941 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
3001 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
3061 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
3121 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
3181 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
3241 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
3301 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
3361 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
3421 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
3481 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
3541 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
3601 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
3661 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
3721 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
3781 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac
3841 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
3901 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
3961 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
4021 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
4081 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
4141 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
4201 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
4261 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
4321 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
4381 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
4441 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
4501 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
4561 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
4621 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
4681 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
4741 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
4801 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
4861 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
4921 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
4981 cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt aatacgactc actatagggg
5041 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
5101 tatacatatg caccatcatc atcatcattc ttctggtctg gtgccacgcg gttctggtat
5161 gaaagaaacc gctgctgcta aattcgaacg ccagcacatg gacagcccag atctgggtac
5221 cgacgacgac gacaaggcca tggctgatat cggatccgga ggtggtggaa gctacaatga
5281 ttttgggaat tacaacaatc agtcttcaaa ttttggaccc atgaagggag gaaattttgg
5341 aggcagaagc tctggcccct atggcggtgg aggccaatac tttgcaaaac cacgaaacca
5401 aggtggctat ggcggttcca gcagcagcag tagctatggc agtggcagaa gatttggggg
5461 cgggctggta ccgaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
5521 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
5581 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggaggccg atcttcctga
5641 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
5701 caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
5761 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
5821 aattattttt gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg
5881 ttacggccag gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg
5941 agaaaaccgc ctcgcggtga tggtgctgcg ctggagtgac ggcagttatc tggaagatca
6001 ggatatgtgg cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac
6061 acaaatcagc gatttccatg ttgccactcg ctttaatgat gatttcagcc gcgctgtact
6121 ggaggctgaa gttcagatgt gcggcgagtt gcgtgactac ctacgggtaa cagtttcttt
6181 atggcagggt gaaacgcagg tcgccagcgg caccgcgcct ttcggcggtg aaattatcga
6241 tgagcgtggt ggttatgccg atcgcgtcac actacgtctg aacgtcgaaa acccgaaact
6301 gtggagcgcc gaaatcccga atctctatcg tgcggtggtt gaactgcaca ccgccgacgg
6361 cacgctgatt gaagcagaag cctgcgatgt cggtttccgc gaggtgcgga ttgaaaatgg
6421 tctgctgctg ctgaacggca agccgttgct gattcgaggc gttaaccgtc acgagcatca
6481 tcctctgcat ggtcaggtca tggatgagca gacgatggtg caggatatcc tgctgatgaa
6541 gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc tgtggtacac
6601 gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa cccacggcat
6661 ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt
6721 aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt cgctggggaa
6781 tgaatcaggc cacggcgcta atcacgacgc gctgtatcgc tggatcaaat ctgtcgatcc
6841 ttcccgcccg gtgcagtatg aaggcggcgg agccgacacc acggccaccg atattatttg
6901 cccgatgtac gcgcgcgtgg atgaagacca gcccttcccg gctgtgccga aatggtccat
6961 caaaaaatgg ctttcgctac ctggagagac gcgcccgctg atcctttgcc aatacgccca
7021 cgcgatgggt aacagtcttg gcggtttcgc taaatactgg caggcgtttc gtcagtatcc
7081 ccgtttacag ggcggcttcg tctgggactg ggtggatcag tcgctgatta aatatgatga
7141 aaacggcaac ccgtggtcgg cttacggcgg tgattttggc gatacgccga acgatcgcca
7201 gttctgtatg aacggtctgg tctttgccga ccgcacgccg catccagcgc tgacggaagc
7261 aaaacaccag cagcagtttt tccagttccg tttatccggg caaaccatcg aagtgaccag
7321 cgaatacctg ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg
7381 taagccgctg gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat
7441 tgaactgcct gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt
7501 agtgcaaccg aacgcgaccg catggtcaga agccgggcac atcagcgcct ggcagcagtg
7561 gcgtctggcg gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct
7621 gaccaccagc gaaatggatt tttgcatcga gctgggtaat aagcgttggc aatttaaccg
7681 ccagtcaggc tttctttcac agatgtggat tggcgataaa aaacaactgc tgacgccgct
7741 gcgcgatcag ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg
7801 cattgaccct aacgcctggg tcgaacgctg gaaggcggcg ggccattacc aggccgaagc
7861 agcgttgttg cagtgcacgg cagatacact tgctgatgcg gtgctgatta cgaccgctca
7921 cgcgtggcag catcagggga aaaccttatt tatcagccgg aaaacctacc ggattgatgg
7981 tagtggtcaa atggcgatta ccgttgatgt tgaagtggcg agcgatacac cgcatccggc
8041 gcggattggc ctgaactgcc agctggcgca ggtagcagag cgggtaaact ggctcggatt
8101 agggccgcaa gaaaactatc ccgaccgcct tactgccgcc tgttttgacc gctgggatct
8161 gccattgtca gacatgtata ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg
8221 gacgcgcgaa ttgaattatg gcccacacca gtggcgcggc gacttccagt tcaacatcag
8281 ccgctacagt caacagcaac tgatggaaac cagccatcgc catctgctgc acgcggaaga
8341 aggcacatgg ctgaatatcg acggtttcca tatggggatt ggtggcgacg actccctgag
8401 cccgtcagta tcggcggaat tccagctgag cgccggtcgc taccattacc agttggtctg
8461 gtctcaaaaa taagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg
8521 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
8581 cataacccct tggggcctct aaacgggtct tga
//
| {
"Vector backbone": "pET30a",
"Backbone size w/o insert (bp)": "5422",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Super-resolved 3D tracking of cargo transport through nuclear pore complexes",
"Publication": "Nat Cell Biol. 2022 Jan;24(1):112-122. doi: 10.1038/s41556-021-00815-6. Epub 2022 Jan 10.",
"DOI": "10.1038/s41556-021-00815-6.",
"Authors": "Chowdhury R, Sau A, Musser SM /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/35013558",
"PubMed ID": "PubMed 35013558"
} | {
"Gene/Insert name": "M9-BetaGal-8C",
"Species": "Escherichia coli",
"Insert Size (bp)": "3363",
"Mutation": "BetaGal has two cysteine mutations (C77A, C1022S), to reduce the number of maleimide reactive sites, and two enzymatic mutations (E538Q, W999L), to reduce Kcat and sugar binding affinity.",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7 Promoter primer",
"3′ sequencing primer": "T7 Terminator primer"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
CMV-d2eGFP-16pf | 21969 | Phil Sharp | Ebert et al Nat Methods. 2007 Sep . 4(9):721-6. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/28266/a21717e9-07bc-423e-9ebd-d515e95cdb87/addgene-plasmid-21969-sequence-28266.gbk | partial | LOCUS sequence-28266-e 1028 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 1028)
AUTHORS Ebert MS, Neilson JR, Sharp PA
TITLE MicroRNA sponges: competitive inhibitors of small RNAs in mammalian
cells.
JOURNAL Nat Methods. 2007 Sep . 4(9):721-6.
PUBMED 17694064
REFERENCE 2 (bases 1 to 1028)
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
Methods. 2007 Sep . 4(9):721-6."
FEATURES Location/Qualifiers
source 1..1028
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 161..182
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(437..454)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 443..667
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(800..819)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gacttcagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
61 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
121 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
181 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagaagctt agccatggct
241 tcccgccgga ggtggaggag caggatgatg gcacgctgcc catgtcttgt gcccaggaga
301 gcgggatgga ccgtcaccct gcagcctgtg cttctgctag gatcaatgtg taggcggccg
361 cctcgagcgc caatatttac gtgctgctac gccaatattt acgtgctgct agggcccgtt
421 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
481 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
541 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
601 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
661 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
721 tgtagcggcg cattaancgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
781 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
841 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
901 cggcacctcg accccaaaaa cttgattagg gtgatggnnc gngtactaga agtcctattc
961 cgagttctat ctctagaant ataggacttc ctggnaaaaa gcctgaactc ncgcgacgtc
1021 tgtcanna
//
| {
"Vector backbone": "pcDNA5",
"Backbone size w/o insert (bp)": "5000",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "MicroRNA sponges: competitive inhibitors of small RNAs in mammalian cells",
"Publication": "Nat Methods. 2007 Sep . 4(9):721-6.",
"DOI": "10.1038/nmeth1079",
"Authors": "Ebert MS, Neilson JR, Sharp PA",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17694064",
"PubMed ID": "PubMed 17694064"
} | {
"Gene/Insert name": "CMV d2eGFP miR-16 perfect",
"Species": "",
"Insert Size (bp)": "50",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (not destroyed)",
"3′ cloning site": "ApaI (not destroyed)",
"5′ sequencing primer": "n/a",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
IOS1 A1_pECIA14 | 114736 | Prey vector IOS1 A1_pECIA14 should be used with bait vector IOS1 A1_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/270064/72e88661-3b0b-42df-96dc-fec4083b1f14/addgene-plasmid-114736-sequence-270064.gbk | full | LOCUS sequence-270064- 6668 bp DNA circular SYN 26-AUG-2024
DEFINITION Prey vector IOS1 A1_pECIA14 should be used with bait vector IOS1
A1_pECIA2..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6668)
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 6668)
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..6668
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(38..60)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 160..179
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 373..395
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 388..404
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 410..836
/label=MT promoter
/note="Drosophila metallothionein promoter"
sig_peptide 854..907
/codon_start=1
/product="Drosophila BiP signal sequence"
/label=BiP signal sequence
/translation="MKLCILLAVVAFVGLSLG"
CDS 2351..2374
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 4049..4072
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 4088..4105
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4128..4145)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4178..4312
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4202..4221)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4256..4275
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4456..4472)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4456..4472)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4469..4491)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4480..4496
/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(4504..4534)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4549..4570
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4687..4704)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4858..5446)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4938..4957)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5617..6477)
/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 6240..6259
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6478..6582)
/gene="bla"
/label=AmpR promoter
primer_bind 6650..6668
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
61 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
121 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
181 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
241 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
301 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
361 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatta attcgttgca
421 ggacaggatg tggtgcccga tgtgactagc tctttgctgc aggccgtcct atcctctggt
481 tccgataaga gacccagaac tccggccccc caccgcccac cgccaccccc atacatatgt
541 ggtacgcaag taagagtgcc tgcgcatgcc ccatgtgccc caccaagagc tttgcatccc
601 atacaagtcc ccaaagtgga gaaccgaacc aattcttcgc gggcagaaca aaagcttctg
661 cacacgtctc cactcgaatt tggagccggc cggcgtgtgc aaaagaggtg aatcgaacga
721 aagacccgtg tgtaaagccg cgtttccaaa atgtataaaa ccgagagcat ctggccaatg
781 tgcatcagtt gtggtcagca gcaaaatcaa gtgaatcatc tcagtgcaac taaagggggg
841 atccgatctc aatatgaagt tatgcatatt actggccgtc gtggcctttg ttggcctctc
901 gctcgggtcg ggattcatca gtttggattg cggttcgcct agagaaacta gctttcgtga
961 gaaaacaaca aacattactt acatttccga tgcgaatttc atcaataccg gtgttggtgg
1021 aagcatcaaa caaggatacc ggactcagtt tcagcaacaa acatggaacc tacggagttt
1081 tccacaaggc ataagaaact gctatacctt aaacctaacc ataggcgacg aatatctaat
1141 cagagccaat ttccttcacg gcggttatga cgacaaaccc tcaacgcaat tcgagctcta
1201 cctcggacct aatctttggt ctacagttac aaccacgaac gagaccgaag catcaatctt
1261 tgagatgatt catatcttga ccactgatcg tttacaaatc tgtctggtca agacaggaaa
1321 cgcaacgcct tttatctccg ctttagagct tcgtaaactc atgaacacaa cttacttgac
1381 tcgacaaggt tccttgcaga ccttcatcag agcagacgtt ggcgcgactg ttaaccaagg
1441 ctacaggtac ggaatcgatg tgtttgatcg tgtctggact ccgtataatt tcggaaactg
1501 gtcacagatt agtaccaatc agtcagtgaa tataaacaat gactatcaac caccagagat
1561 tgctatggtc acagcctctg ttccaacaga cccggacgca gctatgaaca tttcgctagt
1621 cggggtggaa cgtactgtgc agttctacgt ttttatgcat tttgcagaga ttcaagagct
1681 caaatccaat gatacaagag agttcaacat catgtacaat aacaagcata tctatggacc
1741 ttttagacca cttaatttca ccacgtcttc cgtatttaca cctaccgaag ttgtcgccga
1801 tgcaaatggg cagtacatat tttcgcttca aagaacagga aattcgactt tacctcctct
1861 gctcaacgct atggagattt attcggttaa cttgttgccg caacaggaaa ctgatagaaa
1921 agaagttgat gcaatgatga acatcaagtc ggcttacggc gtgaacaaga ttgactggga
1981 aggagatcct tgcgtgccgc tggattacaa gtggtctggt gttaattgca cttatgttga
2041 caatgagacc ccaaagatca tttctttgga tctgtctaca agtggcttga ccggagagat
2101 cttagaattt atatcagacc ttaccagttt agaagttctt gacttgtcta acaattcttt
2161 aactggttca gtcccagagt ttctagctaa tatggaaacc ttaaaactca taaatctatc
2221 tggaaatgag ctaaatggct ctatccctgc aactctcctt gataaagaac gacgaggatc
2281 tattacgtta agtatcgaag gaaataccgg gctttgttca tctacatcat gcgctaccac
2341 gaaaaagaag ctggaggtgc tgttccaggg accaagcggc aatcccggcg gcgatctggc
2401 cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct
2461 gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg agtgcgacgc
2521 ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg aggagaaccc
2581 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
2641 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
2701 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
2761 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
2821 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
2881 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
2941 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
3001 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
3061 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
3121 gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg
3181 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
3241 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
3301 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
3361 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
3421 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
3481 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
3541 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
3601 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
3661 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
3721 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
3781 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
3841 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
3901 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
3961 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
4021 ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata aggccagcgg
4081 caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
4141 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
4201 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
4261 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
4321 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
4381 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
4441 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
4501 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
4561 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
4621 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
4681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
4741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
4801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
4861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
4921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
4981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
5041 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
5101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
5161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
5221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
5281 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
5341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
5401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
5461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
5521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
5581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
5641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
5701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
5761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
5821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
5881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
5941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
6001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
6061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
6121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
6181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
6241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
6301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
6361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
6421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
6481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
6541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
6601 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
6661 cacgaggc
//
| {
"Vector backbone": "pECIA14",
"Backbone size w/o insert (bp)": "",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "An extracellular network of Arabidopsis leucine-rich repeat receptor kinases",
"Publication": "Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. Epub 2018 Jan 10.",
"DOI": "10.1038/nature25184.",
"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 /nature",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29320478",
"PubMed ID": "PubMed 29320478"
} | {
"Gene/Insert name": "AT1G51800",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Metallothionein (Copper-Inducible)",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pHH0103_CLTC_Clathrin | 109997 | Protein expression and purification of CLTC_Clathrin | Sachdev Sidhu | Teyra et al Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/215485/1df52119-1da4-4c40-b9cd-6678718dffc7/addgene-plasmid-109997-sequence-215485.gbk | full | LOCUS sequence-215485- 7815 bp DNA circular SYN 26-AUG-2024
DEFINITION Protein expression and purification of CLTC_Clathrin.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7815)
AUTHORS Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz
AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
TITLE Large-scale survey and database of high affinity ligands for peptide
recognition modules.
JOURNAL Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
PUBMED 33438817
REFERENCE 2 (bases 1 to 7815)
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 Syst
Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: "
10.15252/msb.20199310"
FEATURES Location/Qualifiers
source 1..7815
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..29
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 37..53
/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 42..64
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 138..155
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 171..824
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 782..804
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
CDS 831..854
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 867..884
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
terminator 2010..2057
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin complement(3650..4105)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3787..3808)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3999..4018
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind complement(4147..4169)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4255..4395
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4310..4329
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4410..4427)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4581..5168)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5339..6199)
/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 5962..5981
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6200..6304)
/gene="bla"
/label=AmpR promoter
primer_bind 6372..6390
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6413..6490
/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 6491..7573
/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(6510..6529)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
901 cggccatggc gcagattctg ccgatccgtt ttcaagaaca tctgcagctg caaaacctgg
961 gcattaaccc ggccaatatc ggttttagca ccctgacgat ggaatctgat aaattcattt
1021 gcatccgcga aaaagtgggt gaacaggcgc aagtggttat tatcgatatg aacgacccga
1081 gcaatccgat tcgtcgcccg atcagtgcag actccgctat catgaacccg gcatctaaag
1141 tgattgcgct gaaagccggc aaaaccctgc agatcttcaa catcgaaatg aaaagtaaaa
1201 tgaaagcgca caccatgacg gatgacgtta cgttctggaa atggatttcc ctgaacaccg
1261 ttgcactggt cacggataat gctgtttatc attggtcaat ggagggtgaa tcgcagccgg
1321 tcaaaatgtt tgatcgtcac agctctctgg caggctgcca gattatcaac taccgcaccg
1381 acgctaaaca aaaatggctg ctgctgacgg gcatcagtgc acagcaaaat cgtgtcgtgg
1441 gtgctatgca gctgtatagt gtggatcgca aagtttccca accgattgaa ggtcatgcgg
1501 cctcatttgc gcagttcaaa atggaaggca acgccgaaga atcgaccctg ttttgtttcg
1561 cggtccgtgg tcaggccggc ggtaaactgc acattatcga agttggcacc ccgccgacgg
1621 gtaaccaacc gtttccgaaa aaagccgtgg atgtgttttt cccgccggaa gcgcagaatg
1681 actttccggt ggccatgcaa atcagcgaaa aacatgatgt tgtcttcctg atcaccaaat
1741 atggctacat tcacctgtat gacctggaaa ccggcacgtg catttacatg aatcgtatct
1801 ctggcgaaac cattttcgtt acggcaccgc atgaagcaac cgctggcatt atcggtgtca
1861 accgcaaggg tcaggtgctg agcgtctgtg tggaagaaga aaatattatc ccgtacatta
1921 ccaacgtgct gcagaatccg gatctggcac tgcgtatggc tgttcgcaac aatctggcgg
1981 gcgccgaaga actggcggcc gcataataaa ccgatacaat taaaggctcc ttttggagcc
2041 ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccttt agttgttcct
2101 ttctattctc actccgctga aactgttgaa agttgtttag caaaacccca tacagaaaat
2161 tcatttacta acgtctggaa agacgacaaa actttagatc gttacgctaa ctatgagggc
2221 tgtctgtgga atgctacagg cgttgtagtt tgtactggtg acgaaactca gtgtctagct
2281 agagtggcgg tggctctggt tccggtgatt ttgattatga aaagatggca aacgctaata
2341 agggggctat gaccgaaaat gccgatgaaa acgcgctaca gtctgacgct aaaggcaaac
2401 ttgattctgt cgctactgat tacggtgctg ctatcgatgg tttcattggt gacgtttccg
2461 gccttgctaa tggtaatggt gctactggtg attttgctgg ctctaattcc caaatggctc
2521 aagtcggtga cggtgataat tcacctttaa tgaataattt ccgtcaatat ttaccttccc
2581 tccctcaatc ggttgaatgt cgcccttttg tctttggcgc tggtaaacca tatgaatttt
2641 ctattgattg tgacaaaata aacttattcc gtggtgtctt tgcgtttctt ttatatgttg
2701 ccacctttat gtatgtattt tctacgtttg ctaacatact gcgtaataag gagtcttaat
2761 catgccagtt cttttggcta gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg
2821 cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga
2881 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc
2941 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc
3001 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
3061 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
3121 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
3181 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
3241 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
3301 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
3361 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
3421 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
3481 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
3541 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
3601 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca ggatccgcga aattgtaagc
3661 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
3721 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
3781 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
3841 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
3901 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
3961 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
4021 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
4081 cttaatgcgc cgctacaggg cgcgtcgcgg atcctgcctc gcgcgtttcg gtgatgacgg
4141 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
4201 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
4261 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
4321 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
4381 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
4441 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
4501 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4561 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4621 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca gggtttcccc
4681 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
4741 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
4801 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
4861 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
4921 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
4981 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
5041 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
5101 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
5161 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
5221 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
5281 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
5341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
5401 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
5461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
5521 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
5581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
5641 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
5701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
5761 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
5821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
5881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
5941 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
6001 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
6061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
6121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
6181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
6241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
6301 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
6361 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc ccgacaccat
6421 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
6481 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
6541 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
6601 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
6661 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
6721 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
6781 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
6841 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
6901 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
6961 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
7021 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
7081 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
7141 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
7201 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
7261 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
7321 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
7381 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
7441 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
7501 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
7561 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcacaattct
7621 catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
7681 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
7741 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
7801 ttctgaaatg agctg
//
| {
"Vector backbone": "pHH0103",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Large-scale survey and database of high affinity ligands for peptide recognition modules",
"Publication": "Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.",
"DOI": "10.15252/msb.20199310.",
"Authors": "Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS /msb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33438817",
"PubMed ID": "PubMed 33438817"
} | {
"Gene/Insert name": "CLTC_Clathrin",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "CLTC",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Sfi1 (unknown if destroyed)",
"3′ cloning site": "Not1 (unknown if destroyed)",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
CDS12_mTurquoise2 | 136089 | FP for N-term fusion with a CTAG | Jim Haseloff | Sauret-Gueto et al ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/275020/c70e3173-4f94-417c-b041-bcbf6ec01e77/addgene-plasmid-136089-sequence-275020.gbk | full | LOCUS sequence-275020- 2777 bp DNA circular SYN 26-AUG-2024
DEFINITION FP for N-term fusion with a CTAG.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2777)
AUTHORS Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M,
Markel K, Delmans M, West A, Patron NJ, Haseloff J
TITLE Systematic tools for reprogramming plant gene expression in a simple
model, Marchantia polymorpha.
JOURNAL ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511.
PUBMED 32163700
REFERENCE 2 (bases 1 to 2777)
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. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511."
FEATURES Location/Qualifiers
source 1..2777
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 83..101
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator complement(105..148)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
CDS 165..881
/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(210..231)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(471..490)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 818..839
/label=EGFP-C
/note="EGFP, forward primer"
terminator 901..958
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
primer_bind complement(982..999)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1153..1741)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1233..1252)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(1924..2018)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2039..2698)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 2613..2632
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
1 ttggaacctc ttacgtgccc gatcaactcg agtgccacct gacgtctaag aaaccattat
61 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa
121 atccttagct ttcgctaagg atgatttctg gaattcggtc tcaaatggtg agcaagggcg
181 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
241 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
301 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgt
361 cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
421 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
481 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
541 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
601 actttagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
661 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
721 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
781 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
841 ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggcttcgcga gaccctgcag
901 tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
961 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
1021 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
1081 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
1141 gttgctggcg tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc
1201 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
1261 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
1321 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
1381 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
1441 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
1501 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
1561 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
1621 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
1681 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
1741 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
1801 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
1861 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgaggct
1921 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
1981 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
2041 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
2101 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
2161 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
2221 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
2281 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
2341 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
2401 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
2461 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
2521 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
2581 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
2641 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
2701 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
2761 tttcattatg gtgaaag
//
| {
"Vector backbone": "pUAP4",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant Expression, Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Chloramphenicol, 25 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "OpenMTA",
"Industry Terms": "OpenMTA"
} | {
"Title": "Systematic tools for reprogramming plant gene expression in a simple model, Marchantia polymorpha",
"Publication": "ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511.",
"DOI": "10.1021/acssynbio.9b00511.",
"Authors": "Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M, Markel K, Delmans M, West A, Patron NJ, Haseloff J /acssynbiob",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32163700",
"PubMed ID": "PubMed 32163700"
} | {
"Gene/Insert name": "CDS12_mTurquoise2(noStop)",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "BsaI/ SapI domesticated",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BsaI (destroyed during cloning)",
"3′ cloning site": "BsaI (destroyed during cloning)",
"5′ sequencing primer": "n/a",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pGBW-m4134408 | 152305 | Mammalian expression plasmid for SARS-CoV-2 nsp9 | 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/313244/d6488cdc-96fe-4c29-b97a-fbd96c23d9dc/addgene-plasmid-152305-sequence-313244.gbk | full | LOCUS sequence-313244- 6485 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp9.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6485)
TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for
protein expression and/or purification
REFERENCE 2 (bases 1 to 6485)
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..6485
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 306..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 925..945
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 946..972
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 1025..1146
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1036..1055)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1090..1109
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
terminator 1261..1307
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 1337..1473
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 1518..1589
/label=3' beta-globin insulator
/note="insulator downstream of the human beta-globin locus
(Farrell et al., 2002)"
rep_origin complement(1815..2403)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1895..1914)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2903..4112
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 3949..3968
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 4116..4715
/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(4116..4135)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4612..4632
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4742..4790
/note="synthetic polyadenylation signal"
CDS 5056..5916
/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(5274..5293)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
terminator 5929..6065
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 6217..6288
/label=5' beta-globin insulator
/note="insulator upstream of the human beta-globin locus
(Farrell et al., 2002)"
terminator complement(6293..6339)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
polyA_signal 6357..6405
/note="synthetic polyadenylation signal"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaataa cgagttaagc
601 ccagtagcgc ttcgccagat gagttgcgcc gcaggtacaa cccaaactgc ttgtacggat
661 gacaacgccc tcgcttacta taataccaca aagggaggcc gattcgttct ggccctgttg
721 tcggatctgc aagacttgaa atgggcacgt tttcccaagt ccgacgggac tggcaccatt
781 tacacagaac tagagccgcc atgcaggttc gtgactgata cccctaaagg acccaaggtc
841 aaatatctgt attttatcaa ggggcttaac aatctcaata gaggcatggt gttaggatct
901 ctagccgcaa cggtgcggct gcaagaaaac ctttactttc agtcgtaccc ctatgacgta
961 ccagattatg cgtaagctag cttgactgac tgagatacag cgtaccttca gctcacagac
1021 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
1081 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
1141 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
1201 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
1261 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
1321 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
1381 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
1441 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
1501 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
1561 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
1621 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
1681 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
1741 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
1801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
1861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
1921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
1981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
2041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
2101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
2161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
2221 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
2281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
2341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
2401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
2461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
2521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
2581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
2641 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
2701 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
2761 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
2821 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
2881 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
2941 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
3001 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
3061 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
3121 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
3181 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
3241 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
3301 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
3361 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
3421 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
3481 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
3541 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
3601 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
3661 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
3721 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
3781 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
3841 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
3901 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
3961 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
4021 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
4081 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
4141 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
4201 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
4261 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
4321 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
4381 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
4441 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
4501 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
4561 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
4621 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
4681 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
4741 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
4801 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
4861 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
4921 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
4981 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
5041 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
5101 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
5161 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
5221 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
5281 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
5341 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
5401 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
5461 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
5521 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
5581 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
5641 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
5701 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
5761 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
5821 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
5881 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
5941 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
6001 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
6061 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
6121 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
6181 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
6241 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
6301 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
6361 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
6421 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
6481 ttccg
//
| {
"Vector backbone": "pTwist_CMV_Puro_6",
"Backbone size w/o insert (bp)": "6080",
"Vector type": "Mammalian Expression",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "OpenMTA",
"Industry Terms": "OpenMTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "SARS-CoV-2 nsp9",
"Species": "Severe acute respiratory syndrome coronavirus 2",
"Insert Size (bp)": "338",
"Mutation": "wt",
"Entrez Gene": "ORF1ab",
"Promoter": "CMV",
"Tag / Fusion Protein": "Cleavable TEV;HA (C terminal on backbone)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
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