LOCUS Exported 4532 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-9-1]. ACCESSION . VERSION . KEYWORDS pAWSV-37 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4532) 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 4532) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4532 /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(2347..2365) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2386..2402) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2386..2402) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2399..2421) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2410..2426 /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(2434..2464) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2479..2500 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2617..2634) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2788..3376) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2868..2887) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3547..4407) /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 4170..4189 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4408..4512) /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 cgtgagctga attccgttgc ggatcctgct gggggaaaaa tatggcacct 1321 ggctgaagtg aaatgggact gtgactccta cctgtgccga ctggggtgct gggggggtga 1381 aatgtcgccc gaaccagtga ggtctctctc ctcctcttgt atcctctggt gctggtcagt 1441 cctcgcggct ctagcccctc gctcaagcct ctccctcccg cccgtgaggc cggctttccc 1501 cgggcccctc tgcgcagtgt atggggttat ttttactttc ggttatctag ctttatgaag 1561 actccacacc actcatacag ctagataacc aaagataaca accaaccccg cctcctggct 1621 gctgtcgccg cctctcccac gcagcctccc ggccgccgcc gccgccagca cctccgcagc 1681 ttcccggtcg cccgtcagcg ggagtaggag ggaagggaca cgagtggagt tgagggggag 1741 ggtgaagaga gaaatgaagt ccgagacaaa acaacaacaa aaacctcaga cacggagata 1801 cagacacgac agagaccgaa agatcttcag cctaaagctt tttccccgta tccccccagg 1861 tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt 1921 ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac 1981 cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat 2041 ccctgggggc tttggggggg ggctgtcccc gtgcactcag cctaaagctt tttccccgta 2101 tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc 2161 gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg 2221 agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac 2281 gtaattacat ccctgggggc tttggggggg ggctgtcccc atttaaatca gctccagctt 2341 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc 2401 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 2461 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 2521 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 2581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 2641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 2701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 2761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 2821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 2881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 2941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 3001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 3061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 3121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 3181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 3241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 3301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 3361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 3421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 3481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 3541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 3601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 3661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 3721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 3781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 3841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 3901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 3961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 4021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 4081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 4141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 4201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 4261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 4321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 4381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 4441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 4501 aggggttccg cgcacatttc cccgaaaagt gc //