LOCUS Exported 4493 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-181a]. ACCESSION . VERSION . KEYWORDS pAWSV-14 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4493) 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 4493) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4493 /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(2308..2326) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2347..2363) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2347..2363) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2360..2382) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2371..2387 /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(2395..2425) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2440..2461 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2578..2595) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2749..3337) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2829..2848) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3508..4368) /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 4131..4150 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4369..4473) /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 tataagccga attccgttgc ggatccatca tggactgctc cttaccttgt 1321 tgaaatgaaa cacttttttt gtaaaaacaa acaaataata ataaaacaca caacagaaag 1381 caaggaacag tgagcagtag gaataaaatt cactggacca catttggggg attgcaggac 1441 catttctggt gaactcatgt gacatgtgaa ggtacagctt gctcttttgc ttcaggaaat 1501 gtagtgtcgt ctgtaaggac cccaaggtac agtcaacggt cagtggtttt ttcaaatccc 1561 aaactcaccg acagcgttga atgttccttg gagtcctctg aaggctggcc tgatagccct 1621 tcttcattct cccatccaca tcccaagaac cagtgagcag tgagaccacc gctgatggat 1681 gctcttacat tcctctctgg gtttcaagtc gagagtattt aaattataaa atcaatttgt 1741 gtgcagcaga gaactaatga actgtgggat agactgtgca gagatcttca gcctaaagct 1801 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg 1861 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg 1921 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc 1981 gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc cgtgcactca 2041 gcctaaagct ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag 2101 cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg 2161 ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct 2221 gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc 2281 catttaaatc agctccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta 2341 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 2401 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 2461 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 2521 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 2581 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 2641 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 2701 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 2761 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 2821 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 2881 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 2941 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 3001 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 3061 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 3121 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 3181 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 3241 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 3301 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 3361 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 3421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 3481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 3541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 3601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 3661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 3721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 3781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 3841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 3901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 3961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 4021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 4081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 4141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 4201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 4261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 4321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 4381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 4441 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgc //