LOCUS Exported 4269 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pBT264-[miR-181c]. ACCESSION . VERSION . KEYWORDS pAWSV-15 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4269) 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 4269) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4269 /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(2084..2102) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2123..2139) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2123..2139) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2136..2158) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2147..2163 /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(2171..2201) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2216..2237 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2354..2371) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2525..3113) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2605..2624) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3284..4144) /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 3907..3926 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4145..4249) /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 atttggacga attccgttgc ggatccggaa ggtcagagtc accggcagga 1321 ggatggcaat tccaggcctc agggtccact caacggtcga tggtttgcct gagctgccca 1381 aactcaccga caggttgaat gttcccccaa acccttggca aattttccgg gacctggagt 1441 cgggggtggg gtaggtggca gggaaccaga gagcatgaag agggagagga ggagagagta 1501 aggaacccct tttctctcat gaaataaaca aacgcttttc aggtgctgtt tgcacctaga 1561 tcttcagcct aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc 1621 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc 1681 acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc 1741 gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc 1801 tgtccccgtg cactcagcct aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc 1861 aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg 1921 ctgtccccgc acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc 1981 cgggcggctc gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt 2041 gggggggggc tgtccccatt taaatcagct ccagcttttg ttccctttag tgagggttaa 2101 tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 2161 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 2221 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 2281 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 2341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 2401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 2461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 2521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 2581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 2641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 2701 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 2761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 2821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 2881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 2941 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 3001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 3061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 3121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 3181 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 3241 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 3301 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 3361 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 3421 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 3481 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 3541 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 3601 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 3661 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 3721 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 3781 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 3841 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 3901 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 3961 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 4021 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 4081 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 4141 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 4201 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 4261 gaaaagtgc //