LOCUS V022178 6891 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022178 VERSION V022178 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 647..1561 /label="Ubb(NM_011664)" /note="Ubb(NM_011664)" /gene="Ubb" CDS 1562..1579 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" misc_feature 1667..2255 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2321..2369 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2571..2999 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3013..3342 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3409..4203 /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 4379..4500 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4549..4565) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4573..4589 /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(4597..4627) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4642..4663 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4951..5539) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5710..6570) /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(6571..6675) /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 gccaccatgc agatcttcgt 661 gaagaccctg accggcaaga ccatcaccct agaggtggag cccagtgaca ccatcgagaa 721 cgtgaaggcc aagatccagg ataaagaggg catcccccct gaccagcaga ggctgatctt 781 tgccggcaag cagctggaag atggccgcac cctctctgat tacaacatcc agaaggagtc 841 aaccctgcac ctggtccttc gcctgagagg tggcatgcag atcttcgtga agaccctgac 901 cggcaagacc atcaccctgg aggtggagcc cagtgacacc atcgagaacg tgaaggccaa 961 gatccaggat aaagagggca tcccccctga ccagcagagg ctgatctttg ccggcaagca 1021 gctggaagat ggccgcaccc tctctgatta caacatccag aaagagtcaa ccctgcacct 1081 ggtcctccgt ctgaggggtg gcatgcagat cttcgtgaag accctgactg gcaagaccat 1141 caccctggag gtggagccca gtgacaccat cgagaacgtg aaggccaaga tccaggataa 1201 agagggcatc ccccctgacc agcagaggct gatctttgcc ggcaagcagc tggaagatgg 1261 ccgcaccctc tctgattaca acatccagaa ggagtcaacc ctgcacctgg tccttcgcct 1321 gagaggtggc atgcagatct tcgtgaagac cctgaccggc aagaccatca ccttggaggt 1381 ggagcccagt gacaccatcg agaatgtgaa ggccaagatc caggataaag agggcatccc 1441 ccctgaccag cagaggctga tctttgccgg caagcagcta gaagatggcc gcactctctc 1501 tgattacaac atccagaaag agtcgaccct gcacctggtc ctccgtctga ggggtggcta 1561 tcaccatcac catcaccatt aggatccact agtccagtgt ggtggaattc tgcagatatc 1621 cagcacagtg gcggccgctc gagtctagag ggcccgatat ctcgacaatc aacctctgga 1681 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 1741 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 1801 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 1861 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 1921 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 1981 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 2041 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 2101 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 2161 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 2221 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa 2281 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 2341 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca 2401 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc 2461 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc 2521 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg 2581 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 2641 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 2701 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg 2761 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 2821 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 2881 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 2941 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 3001 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 3061 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 3121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 3181 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 3241 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 3301 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 3361 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 3421 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 3481 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 3541 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 3601 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 3661 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 3721 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 3781 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 3841 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 3901 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 3961 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 4021 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 4081 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 4141 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 4201 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 4261 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 4321 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa 4381 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 4441 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 4501 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt 4561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 4621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 4681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 4741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 4801 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 4861 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 4921 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 4981 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 5041 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 5101 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag 5161 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 5221 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5281 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5341 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 5401 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 5461 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 5521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 5581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 5641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 5701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga 6721 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc 6781 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa 6841 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g //