LOCUS 40924_10176 6011 bp DNA circular SYN 08-JAN-2019 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6011) TITLE Direct Submission REFERENCE 2 (bases 1 to 6011) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..6011 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 50..429 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 430..633 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 787..1375 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" polyA_signal 1441..1489 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 1691..2119 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2133..2462 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2529..3320 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 3499..3632 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3669..3685) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3693..3709) /label=lac operator /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(3717..3747) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3762..3783) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4071..4659) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4833..5690) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5691..5795) /label=AmpR promoter ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagg atcgaaccct taagggttcg atccctaccg gttagtaatg agtttgatat 781 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 841 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 901 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 961 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 1021 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 1081 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 1141 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 1201 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 1261 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 1321 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga 1381 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 1441 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg 1501 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac 1561 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc 1621 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg 1681 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 1741 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 1801 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg 1861 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 1921 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 1981 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2041 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa 2101 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2161 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2221 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2281 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 2341 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 2401 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 2461 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 2521 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 2581 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 2641 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 2701 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 2761 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 2821 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 2881 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 2941 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3001 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 3061 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3121 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 3181 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 3241 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 3301 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg 3361 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc 3421 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 3481 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat 3541 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 3601 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg 3661 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 3721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 3781 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 3841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 3901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 3961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 4021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 4081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 4141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 4201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 4261 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 4321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 4381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 4441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 4501 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 4561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 4621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 4681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 4741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 4801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 4861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 4921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 4981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 5041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 5101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 5161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 5221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 5281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 5341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 5401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 5461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 5521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 5581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 5641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 5701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 5761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 5821 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct 5881 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt 5941 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga 6001 atctgcttag g //