LOCUS Exported 4427 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-216]. ACCESSION . VERSION . KEYWORDS pAWSV-21 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4427) 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 4427) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4427 /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(2242..2260) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2281..2297) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2281..2297) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2294..2316) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2305..2321 /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(2329..2359) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2374..2395 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2512..2529) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2683..3271) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2763..2782) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3442..4302) /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 4065..4084 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4303..4407) /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 cagtcaccga attccgttgc ggatccggtt tcctttccag cacaacagca 1321 ctttagaaaa agactttact aagattaaaa gaggttgagc atgaaacagt aaccatcgaa 1381 acttttcccc ccgtcttggt catgaaatag aaatttggta tttaattttg gttgccatgc 1441 aagtcttttg attcgtgagg gctaggaaat tgctctgttt agcataatcc cagagaccac 1501 tgtgagggat tgtatgaaca tctcacagtt gccagctgag attaagccaa ctcacagcca 1561 tccgtgttag acctgcaaga gaaagtgtat gagcagaggc acgtggctgc ctcaggtgga 1621 agcacatctt catacatcga gtgcaccagc agaagccctg atgtacccct ctgcattttt 1681 cttgaggcga cattatcaag ctactcctct tgctgagatc ttcagcctaa agctttttcc 1741 ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg 1801 atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg 1861 gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga 1921 gggacgtaat tacatccctg ggggctttgg gggggggctg tccccgtgca ctcagcctaa 1981 agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca 2041 gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct 2101 cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc 2161 tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg tccccattta 2221 aatcagctcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg 2281 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 2341 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 2401 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 2461 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 2521 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 2581 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 2641 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 2701 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 2761 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 2821 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 2881 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2941 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 3001 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 3061 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 3121 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 3181 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 3241 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 3301 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 3361 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 3421 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 3481 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 3541 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 3601 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 3661 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 3721 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 3781 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 3841 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 3901 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 3961 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 4021 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 4081 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 4141 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 4201 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 4261 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 4321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 4381 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgc //