LOCUS Exported 4455 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-488]. ACCESSION . VERSION . KEYWORDS pAWSV-34 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4455) AUTHORS Wong AS, Choi GC, Cheng AA, Purcell O, Lu TK TITLE Massively parallel high-order combinatorial genetics in human cells. JOURNAL Nat Biotechnol. 2015 Sep;33(9):952-61. doi: 10.1038/nbt.3326. Epub 2015 Aug 17. PUBMED 26280411 REFERENCE 2 (bases 1 to 4455) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4455 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(6..461) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(143..164) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 355..374 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 588..610 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 602..619 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 603..619 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 626..645 /label=T7 /note="T7 promoter, forward primer" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 1175..1256 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 1200..1219 /label=SV40pA-R /note="SV40 polyA, reverse primer" promoter complement(2270..2288) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2309..2325) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2309..2325) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2322..2344) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2333..2349 /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(2357..2387) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2402..2423 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2540..2557) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2711..3299) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2791..2810) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3470..4330) /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 4093..4112 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4331..4435) /gene="bla" /label=AmpR promoter ORIGIN 1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct 481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtacagg 661 cgcgcctcag cctaaagctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc 721 agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc 781 cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg 841 ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg 901 ggctgtcccc gtgcactcag cctaaagctt tttccccgta tccccccagg tgtctgcagg 961 ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc 1021 gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag 1081 ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc 1141 tttggggggg ggctgtcccc actagtgatt cgaggcagtg aaaaaaatgc tttatttgtg 1201 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca 1261 acaacaattg aacgcgaaga attccgttgc ggatcccctc aactacactg cccccaatcc 1321 ttgcctagct caaacccaag gccaatgtgg tagaggaagc aggggtacat atagtatcca 1381 tcttttcaca tacaacacac tttattaact ttgtttgtct gaacagcaaa atatgactcc 1441 tgagtcttcc agaagaaatt gagagtcatc tgaccaagaa atagcctttc aaacaatttg 1501 gaaacttgtt tgagagtgcc attatctggg agagatgatt ctctgaaatt ttccactcta 1561 ggctcttctt tcttccttag gaggaaacac cagttgtggt ccaaaaattc ttcttgcaga 1621 aactggtttt gttccataga gcccacaacc caacccaaca cctcttcgcc ttcccccgcc 1681 tccctcactc ccagatcctt taccaccttc tgccacctct atcattccta cctctactac 1741 gccagatctt cagcctaaag ctttttcccc gtatcccccc aggtgtctgc aggctcaaag 1801 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt 1861 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg 1921 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg 1981 gggggctgtc cccgtgcact cagcctaaag ctttttcccc gtatcccccc aggtgtctgc 2041 aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg 2101 cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg 2161 gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg 2221 ggctttgggg gggggctgtc cccatttaaa tcagctccag cttttgttcc ctttagtgag 2281 ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 2341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 2401 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 2461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 2521 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 2581 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 2641 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 2701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 2761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 2821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 2881 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 2941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 3001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 3061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 3121 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 3181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 3241 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 3301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 3361 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 3421 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 3481 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 3541 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 3601 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 3661 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 3721 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 3781 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 3841 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 3901 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 3961 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 4021 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 4081 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 4141 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 4201 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 4261 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 4321 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 4381 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 4441 ttccccgaaa agtgc //