LOCUS V047118 7041 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047118 VERSION V047118 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" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 756..1721 /label="2310022B05Rik(NM_175149)" /note="2310022B05Rik(NM_175149)" /gene="2310022B05Rik" CDS 1728..1793 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1817..2405 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2471..2519 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2721..3149 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3163..3492 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3559..4353 /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 4529..4650 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4699..4715) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4723..4739 /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(4747..4777) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4792..4813 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5101..5689) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5860..6720) /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(6721..6825) /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 gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc gtcgatggcg gcggcgatcg 781 cggcctcgcg ttcggccgtc atgagcggga accggccgct ggacgaccgg gagcggaagc 841 gcttcaccta cttctcatcg ctgagcccca tggccaggaa gatcatgcag gacaaggaga 901 agatccggga gaagtacggg cccgagtggg cgcggctgcc gcccgcgcag caggacgaaa 961 tcatcgaccg ctgcctggtg gggcctcgcg ccccggcggc cgccgccgat gcgggggacg 1021 tgcgggaccc ggcgcgcttc cctggcctgc gcgggcccac gggccagaag ctggtgcgct 1081 tcggcgacga ggacataact tggcaagatg aacactcggc cccattctcc tgggagacga 1141 ggagtcagat ggaattcagc atctcctcct tatccatcca ggagcccagc gctgccactg 1201 tgaccagtga tgccaggtcc ctggccaagg ctcctcaggg cacacagggt tccaaacctg 1261 cccagagcag cagatcctct agcctggatg ccctgggacc ggctcggaaa gaggaagaag 1321 cacccttctg gaagatcaat gccgagcgct ctcgggaggg gcctgaggct gagttccagt 1381 ccctgacacc cagccagatc aagtccatgg agaaggggga aaaggttctg ccagcttgct 1441 accgccaaga gcccaccacg aaagacaggg aggccaaacc tgtgccacag gaacagcaga 1501 ccctccccag tgtgagcgca gagcaagagg tgccccagcc ggtgcaagcc cctgccagct 1561 tgctgcccaa ggctaccccc acggagtccc cagagaagcc accgcctcct gccgtccagc 1621 gggacgaaga tgatgatgct ctgttctcag agcctgcact tgcgcagatc agctccagta 1681 atgtgctctt gaagacaggc tttgattttc tggacaactg gggatccgac tacaaagacc 1741 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag 1801 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 1861 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 1921 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 1981 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 2041 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 2101 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 2161 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc 2221 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta 2281 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 2341 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 2401 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc 2461 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca 2521 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg 2581 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg 2641 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg 2701 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2761 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2821 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat 2881 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2941 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 3001 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 3061 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga 3121 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 3181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3241 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3301 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3361 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3421 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3481 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 3541 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 3601 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 3661 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 3721 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 3781 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 3841 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 3901 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 3961 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 4021 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 4081 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 4141 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 4201 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 4261 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 4321 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac 4381 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4441 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4501 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 4561 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 4621 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 4681 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 4741 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 4801 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 4861 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 4921 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 4981 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5041 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5101 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5161 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5221 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5281 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5341 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 5401 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 5461 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 5521 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 5581 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 5641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 5701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 5761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 5821 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 5881 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 5941 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6001 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6061 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6121 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6181 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6241 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 6301 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 6361 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 6421 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 6481 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 6541 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 6601 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 6661 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 6721 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 6781 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 6841 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt 6901 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag 6961 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat 7021 tgcatgaaga atctgcttag g //