LOCUS V034787 7693 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034787 VERSION V034787 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 964..3183 /label="POLI(NM_007195)" /note="POLI(NM_007195)" /gene="POLI" CDS 3190..3255 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3293..3517 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3563..3991 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4005..4334 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4401..5195 /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 5369..5490 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5539..5555) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5563..5579 /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(5587..5617) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5632..5653 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5941..6526) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6697..7557) /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(7558..7662) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatggaga agctgggggt ggagccggag gaggaaggcg gcggcgacga cgacgaggaa 1021 gacgccgagg cctgggccat ggaactggcg gacgtggggg cggcagccag ctcgcaggga 1081 gttcatgatc aagtgttgcc cacaccaaat gcttcatcca gagtcatagt acatgtggat 1141 ctggattgct tttatgcaca agtagaaatg atctcaaatc cagagctaaa agacaaacct 1201 ttaggggttc aacagaaata tttggtggtt acctgcaact atgaagctag gaaacttgga 1261 gttaagaaac ttatgaatgt cagagatgca aaagaaaagt gtccacagtt ggtattagtt 1321 aatggagaag acctgacccg ctacagagaa atgtcttata aggttacaga attactggaa 1381 gaatttagtc cagttgttga gagacttgga tttgatgaaa attttgtgga tctaacagaa 1441 atggttgaga agagactaca gcagctgcaa agtgatgaac tttctgcggt gactgtgtcg 1501 ggtcatgtat acaataatca gtctataaac ctgcttgacg tcttgcacat cagactactt 1561 gttggatctc agattgcagc agagatgcgg gaagccatgt ataatcagtt ggggctcact 1621 ggctgtgctg gagtggcttc taataaactg ttggcaaaat tagtttctgg tgtctttaaa 1681 ccaaatcaac aaacagtctt attacctgaa agttgtcaac atcttattca tagtttgaat 1741 cacataaagg aaatacctgg tattggctat aaaactgcca aatgtcttga agcactgggt 1801 atcaatagtg tgcgtgatct ccaaaccttt tcacccaaaa ttttagaaaa agaattagga 1861 atttcagttg ctcagcgtat ccaaaagctc agttttggag aggataactc ccctgtgata 1921 ctctcaggac cacctcagtc ctttagtgaa gaagattcat ttaaaaaatg ttcatctgaa 1981 gttgaagcta aaaataagat tgaagaacta cttgctagtc ttttaaacag agtatgccaa 2041 gatggaagga agcctcatac agtgagatta ataatccgtc ggtattcctc tgagaagcac 2101 tatggtcgtg agagtcgtca gtgccctatt ccttcacatg taattcagaa attagggaca 2161 ggaaattatg atgtgatgac cccaatggtt gatatactta tgaaactttt tcgaaatatg 2221 gtgaatgtga agatgccatt tcaccttacc cttctaagtg tgtgcttctg caaccttaaa 2281 gcactaaata ctgctaagaa agggcttatt gattattatt taatgccatc attatcaact 2341 acttcacgct ctggcaagca cagttttaaa atgaaagaca ctcatatgga agattttccc 2401 aaagacaaag aaacaaaccg ggatttccta ccaagtggaa gaattgaaag tacaagaact 2461 agggagtctc cactagatac cacaaatttt tctaaagaaa aagacattaa tgaattccca 2521 ctctgttcac ttcctgaagg tgttgaccaa gaagtcttca agcagcttcc agtagatatt 2581 caagaagaaa tcctttctgg aaaatctagg gaaaaatttc aagggaaagg aagtgtgagt 2641 tgtccattac atgcctctag aggagtatta tctttctttt ctaaaaaaca aatgcaagat 2701 attcccataa atcctagaga tcatttatcc agtagcaaac aggtatcctc tgtatctcct 2761 tgtgaaccgg gaacatcagg ctttaatagc agtagttctt cttacatgtc tagccaaaag 2821 gattattcat attatttaga taatagatta aaagatgaac gaataagtca aggacctaaa 2881 gaacctcaag gattccactt tacaaattca aaccctgctg tgtctgcttt tcattcattt 2941 ccaaacttgc agagtgagca acttttctcc agaaaccaca ctacagatag ccataagcaa 3001 acagtagcaa cagactctca tgaaggactt acagaaaata gagagccaga ttctgttgat 3061 gagaaaatta ctttcccttc tgacattgat cctcaagttt tctatgaact accagaagca 3121 gtacaaaagg aactgctggc agagtggaag agagcaggat cagatttcca cattggacat 3181 aaactcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag 3241 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct 3301 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 3361 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 3421 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 3481 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 3541 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 3601 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 3661 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 3721 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 3781 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 3841 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 3901 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 3961 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 4021 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 4081 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 4141 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 4201 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 4261 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 4321 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 4381 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 4441 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 4501 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 4561 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 4621 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 4681 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 4741 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 4801 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 4861 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 4921 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4981 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 5041 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 5101 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 5161 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 5221 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 5281 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 5341 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 5401 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 5461 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 5521 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5581 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5641 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5701 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5761 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5821 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6121 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6361 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 6421 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt 6481 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6541 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6601 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6661 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6721 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6781 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6841 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6901 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6961 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 7021 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 7081 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 7141 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 7201 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 7261 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 7321 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7381 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7441 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7501 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 7561 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7621 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7681 gccacctgac gtc //