LOCUS Exported 4456 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-373]. ACCESSION . VERSION . KEYWORDS pAWSV-30 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4456) 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 4456) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4456 /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(2271..2289) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2310..2326) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2310..2326) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2323..2345) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2334..2350 /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(2358..2388) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2403..2424 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2541..2558) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2712..3300) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2792..2811) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3471..4331) /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 4094..4113 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4332..4436) /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 tggagataga attccgttgc ggatccctgc aggtgaaccc cgtatcctgc 1321 ccaccccaga atatgccaat ttttattatc taaaatggag catacagact ggtgccccgc 1381 agagaagccc gggaactcca ggtcatagct gaagctgcag tcagacccgc aggctccctt 1441 gaagcaggat ggttcgtgat gccctactca aacagggaca ccccaaaatc gaagcacttc 1501 ccagtacaga caaaaaggaa agcgccccca ttttgagtat cccagtatga gctcctcgcg 1561 aggatctgcc atcactgtga ctttcttgcc tacaagaggt tggcctccaa tcatccaagc 1621 acatgttaaa tgtcagcaat aaacttgctt gctatggggg aagaggaaag ttacatatca 1681 attataacac caacagcaaa aagcattcct ttctttgtgc atacagcccc ttggtcacag 1741 ctgcagatct tcagcctaaa gctttttccc cgtatccccc caggtgtctg caggctcaaa 1801 gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg 1861 tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg 1921 gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg 1981 ggggggctgt ccccgtgcac tcagcctaaa gctttttccc cgtatccccc caggtgtctg 2041 caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt 2101 gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc 2161 ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg 2221 gggctttggg ggggggctgt ccccatttaa atcagctcca gcttttgttc cctttagtga 2281 gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 2341 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 2401 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 2461 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 2521 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 2581 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 2641 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 2701 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 2761 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 2821 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 2881 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 2941 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 3001 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 3061 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 3121 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 3181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 3241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 3301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 3361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 3421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 3481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 3541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 3601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 3661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 3721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 3781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 3841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 3901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 3961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 4021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 4081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 4141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 4201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 4261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 4321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 4381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 4441 tttccccgaa aagtgc //