LOCUS Exported 4645 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-429]. ACCESSION . VERSION . KEYWORDS pAWSV-32 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4645) 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 4645) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4645 /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(2460..2478) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2499..2515) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2499..2515) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2512..2534) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2523..2539 /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(2547..2577) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2592..2613 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2730..2747) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2901..3489) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2981..3000) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3660..4520) /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 4283..4302 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4521..4625) /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 ctgaatgaga attccgttgc ggatccctcc gggtatctgt gactgtgacc 1321 ccaggggcca ggctgatagc acggggtcca cagcatctgc ccggggacta cagctcgagg 1381 gccccaggag gccgggtggg cacaggccat ggcgagggtg gggcacaaga gccccagacc 1441 ccggcggctt tgcactgatg ggctgcggat gggcacaggc catagtgagg ggggcatgag 1501 agccccagac cgggcggctt tgcactgatg agctgcaggg caggcagctc tctcctctaa 1561 cggtgatcag gcagcggatg gacggtttta ccagacagta ttagacagag ggccaggtct 1621 aaccatgtct ggtaagacgc ccatcggccg gcggtggtgt gggtggcctc cgggtcctgg 1681 gctggtgtct gcggggcctg ngaagggaag gccccggggg cggctcctgc cacctcctgc 1741 agctgccctg atccctcctg ggtctgcaga acgagccccc gaggcacttc cctgacagcc 1801 cccatcccag cacctggtcc cagcccagcc tggctgtgtg ggggtacccg ggagccacct 1861 ggtcacccag gccctgctct tggcaaaggt tccctgcagc ccctcgctgg cttcctgaag 1921 ctcggtcggg ttaagatctt cagcctaaag ctttttcccc gtatcccccc aggtgtctgc 1981 aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg 2041 cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg 2101 gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg 2161 ggctttgggg gggggctgtc cccgtgcact cagcctaaag ctttttcccc gtatcccccc 2221 aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac 2281 cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg 2341 gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta 2401 catccctggg ggctttgggg gggggctgtc cccatttaaa tcagctccag cttttgttcc 2461 ctttagtgag ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga 2521 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 2581 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 2641 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 2701 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 2761 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 2821 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 2881 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 2941 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 3001 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 3061 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 3121 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 3181 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 3241 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 3301 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 3361 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 3421 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 3481 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 3541 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 3601 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 3661 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 3721 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 3781 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 3841 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 3901 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 3961 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 4021 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 4081 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 4141 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 4201 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 4261 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 4321 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 4381 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 4441 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 4501 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 4561 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 4621 ccgcgcacat ttccccgaaa agtgc //