LOCUS V027071 5966 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027071 VERSION V027071 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..1456 /label="ACYP2(NM_001320586)" /note="ACYP2(NM_001320586)" /gene="ACYP2" CDS 1463..1528 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1566..1790 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1836..2264 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2278..2607 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2674..3468 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 3642..3763 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3812..3828) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3836..3852 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(3860..3890) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3905..3926 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4214..4799) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4970..5830) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(5831..5935) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgttgctca cacaaagcct 961 gtttggtggt ctcttcccac ggacgcgcga gacaatgagg agatacaagg tctcgctgtt 1021 ctacctaggc tgttctagaa ctcctaatgt caagctatcc tcctgcctcg gcctcccatg 1081 ctgttgggat tacagctata aattcataca attatcagag tttggttttg gtcaagtcat 1141 aattgtgagt gaagaaccat ggaaggagaa catttcttgc tcatcaacta ctttcataaa 1201 atcaacaatt tgcttaagtg tttgcttcag aatgtataca gaagatgaag ctaggaaaat 1261 aggagtggtt ggctgggtga agaataccag caaaggcacc gtgacaggcc aagtgcaggg 1321 gccagaagac aaagtcaatt ccatgaagtc ctggctgagc aaggttggaa gccctagttc 1381 tcgcattgac cgcacaaact tttctaatga aaaaaccatc tctaagcttg aatactctaa 1441 ttttagtatt agatacctcg aggactacaa agaccatgac ggtgattata aagatcatga 1501 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 1561 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 1621 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 1681 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 1741 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 1801 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 1861 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 1921 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 1981 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 2041 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 2101 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 2161 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 2221 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 2281 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2341 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 2401 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 2461 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 2521 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 2581 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 2641 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 2701 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 2761 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 2821 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 2881 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 2941 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3001 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3061 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 3121 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 3181 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 3241 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 3301 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 3361 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 3421 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 3481 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 3541 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 3601 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 3661 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 3721 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 3781 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 3841 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 3901 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 3961 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4021 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4081 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4141 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4201 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4261 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4321 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4381 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4441 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4501 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4561 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4621 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 4681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 4741 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 4801 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 4861 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 4921 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 4981 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5041 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5101 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5161 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 5221 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 5281 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 5341 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 5401 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 5461 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 5521 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 5581 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 5641 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 5701 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 5761 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 5821 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 5881 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 5941 ttccccgaaa agtgccacct gacgtc //