LOCUS Exported 4432 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-29b]. ACCESSION . VERSION . KEYWORDS pAWSV-24 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4432) 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 4432) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4432 /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(2247..2265) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2286..2302) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2286..2302) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2299..2321) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2310..2326 /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(2334..2364) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2379..2400 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2517..2534) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2688..3276) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2768..2787) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3447..4307) /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 4070..4089 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4308..4412) /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 tcacagatga attccgttgc ggatccgcca gtgcagagac ctgactgcca 1321 tttgtgatat atgccaccac tgttggcagg ggtgaggaag gggtcagcta catgtgaggc 1381 aggttctcac agccatcctg ctgttgcttt caaatacttc agagctgtcc cattcacttc 1441 ttttgctgtt ggtagtgcag cgcagctggt ctcccccaag aacactgatt tcaaatggtg 1501 ctagacaatc actatttaaa tctaaaccac catatgaaac cagcttcctg aagaagcttt 1561 atgatctatt tttttcttca taatgctctc ttacaataaa cccaagacaa cctagaaagg 1621 aaaagttgaa acctagctgt gagttactca gtggtctaaa cagagctttg gcaaatttca 1681 tatattcata aactgatgtc actgaatttg ttattgatgg agatcttcag cctaaagctt 1741 tttccccgta tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga 1801 aagcgatccc gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg 1861 atgcgggggg agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg 1921 ggggagggac gtaattacat ccctgggggc tttggggggg ggctgtcccc gtgcactcag 1981 cctaaagctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc agcgagaagc 2041 gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc 2101 cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg ctcgctgctg 2161 ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg ggctgtcccc 2221 atttaaatca gctccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa 2281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 2341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 2401 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 2461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 2521 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 2581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 2641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 2701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 2761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 2821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 2881 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 2941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 3001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 3061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 3121 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 3181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 3241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 3301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 3361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 3421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 3481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 3541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 3601 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 3661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 3721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 3781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 3841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 3901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 3961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 4021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 4081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 4141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 4201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 4261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 4321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 4381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gc //