LOCUS V027818 8139 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027818 VERSION V027818 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 706..2865 /label="ACSL3(NM_004457)" /note="ACSL3(NM_004457)" /gene="ACSL3" CDS 2872..2895 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" misc_feature 2915..3503 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3569..3617 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3819..4247 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4261..4590 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4657..5451 /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 5627..5748 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5797..5813) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5821..5837 /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(5845..5875) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5890..5911 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6199..6787) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6958..7818) /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(7819..7923) /gene="bla" /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 gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgaa taaccacgtg 721 tcttcaaaac catctaccat gaagctaaaa cataccatca accctattct tttatatttt 781 atacattttc taatatcact ttatactatt ttaacataca ttccgtttta ttttttctcc 841 gagtcaagac aagaaaaatc aaaccgaatt aaagcaaagc ctgtaaattc aaaacctgat 901 tctgcataca gatctgttaa tagtttggat ggtttggctt cagtattata ccctggatgt 961 gatactttag ataaagtttt tacatatgca aaaaacaaat ttaagaacaa aagactcttg 1021 ggaacacgtg aagttttaaa tgaggaagat gaagtacaac caaatggaaa aatttttaaa 1081 aaggttattc ttggacagta taattggctt tcctatgaag atgtctttgt tcgagccttt 1141 aattttggaa atggattaca gatgttgggt cagaaaccaa agaccaacat cgccatcttc 1201 tgtgagacca gggccgagtg gatgatagct gcacaggcgt gttttatgta taattttcag 1261 cttgttacat tatatgccac tctaggaggt ccagccattg ttcatgcatt aaatgaaaca 1321 gaggtgacca acatcattac tagtaaagaa ctcttacaaa caaagttgaa ggatatagtt 1381 tctttggtcc cacgcctgcg gcacatcatc actgttgatg gaaagccacc gacctggtcc 1441 gagttcccca agggcatcat tgtgcatacc atggctgcag tggaggccct gggagccaag 1501 gccagcatgg aaaaccaacc tcatagcaaa ccattgccct cagatattgc agtaatcatg 1561 tacacaagtg gatccacagg acttccaaag ggagtcatga tctcacatag taacattatt 1621 gctggtataa ctgggatggc agaaaggatt ccagaactag gagaggaaga tgtctacatt 1681 ggatatttgc ctctggccca tgttctagaa ttaagtgctg agcttgtctg tctttctcac 1741 ggatgccgca ttggttactc ttcaccacag actttagcag atcagtcttc aaaaattaaa 1801 aaaggaagca aaggggatac atccatgttg aaaccaacac tgatggcagc agttccggaa 1861 atcatggatc ggatctacaa aaatgtcatg aataaagtca gtgaaatgag tagttttcaa 1921 cgtaatctgt ttattctggc ctataattac aaaatggaac agatttcaaa aggacgtaat 1981 actccactgt gcgacagctt tgttttccgg aaagttcgaa gcttgctagg gggaaatatt 2041 cgtctcctgt tgtgtggtgg cgctccactt tctgcaacca cgcagcgatt catgaacatc 2101 tgtttctgct gtcctgttgg tcagggatac gggctcactg aatctgctgg ggctggaaca 2161 atttccgaag tgtgggacta caatactggc agagtgggag caccattagt ttgctgtgaa 2221 atcaaattaa aaaactggga ggaaggtgga tactttaata ctgataagcc acaccccagg 2281 ggtgaaattc ttattggggg ccaaagtgtg acaatggggt actacaaaaa tgaagcaaaa 2341 acaaaagctg atttctttga agatgaaaat ggacaaaggt ggctctgtac tggggatatt 2401 ggagagtttg aacccgatgg atgcttaaag attattgatc gtaaaaagga ccttgtaaaa 2461 ctacaggcag gggaatatgt ttctcttggg aaagtagagg cagctttgaa gaatcttcca 2521 ctagtagata acatttgtgc atatgcaaac agttatcatt cttatgtcat tggatttgtt 2581 gtgccaaatc aaaaggaact aactgaacta gctcgaaaga aaggacttaa agggacttgg 2641 gaggagctgt gtaacagttg tgaaatggaa aatgaggtac ttaaagtgct ttccgaagct 2701 gctatttcag caagtctgga aaagtttgaa attccagtaa aaattcgttt gagtcctgaa 2761 ccgtggaccc ctgaaactgg tctggtgaca gatgccttca agctgaaacg caaagagctt 2821 aaaacacatt accaggcgga cattgagcga atgtatggaa gaaaactcga gcaccatcac 2881 catcaccatc accattaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg 2941 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 3001 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 3061 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 3121 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 3181 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 3241 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 3301 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 3361 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 3421 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 3481 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag 3541 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg 3601 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac 3661 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc 3721 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat 3781 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 3841 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 3901 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 3961 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 4021 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 4081 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 4141 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc 4201 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 4261 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4321 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 4381 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 4441 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 4501 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 4561 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 4621 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 4681 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 4741 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 4801 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 4861 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4921 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4981 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 5041 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 5101 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 5161 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 5221 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 5281 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 5341 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 5401 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 5461 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 5521 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 5581 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 5641 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 5701 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 5761 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 5821 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 5881 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 5941 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 6001 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 6061 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 6121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 6181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 6241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 6301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 6361 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 6421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6601 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 6661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 7021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 7081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 7141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 7201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 7261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 7321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 7381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 7441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 7501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 7561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 7621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 7681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 7741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 7801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 7861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7921 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc 7981 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc 8041 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg 8101 caaggcttga ccgacaattg catgaagaat ctgcttagg //