LOCUS V019270 6941 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019270 VERSION V019270 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 683..1621 /label="MGLL(NM_007283)" /note="MGLL(NM_007283)" /gene="MGLL" CDS 1628..1693 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1717..2305 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2371..2419 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2621..3049 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3063..3392 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3459..4253 /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 4429..4550 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4599..4615) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4623..4639 /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(4647..4677) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4692..4713 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5001..5589) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5760..6620) /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(6621..6725) /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 ggatccgcca ccatggaaac aggacctgaa gacccttcca gcatgccaga 721 ggaaagttcc cccaggcgga ccccgcagag cattccctac caggacctcc ctcacctggt 781 caatgcagac ggacagtacc tcttctgcag gtactggaaa cccacaggca cacccaaggc 841 cctcatcttt gtgtcccatg gagccggaga gcacagtggc cgctatgaag agctggctcg 901 gatgctgatg gggctggacc tgctggtgtt cgcccacgac catgttggcc acggacagag 961 cgaaggggag aggatggtag tgtctgactt ccacgttttc gtcagggatg tgttgcagca 1021 tgtggattcc atgcagaaag actaccctgg gcttcctgtc ttccttctgg gccactccat 1081 gggaggcgcc atcgccatcc tcacggccgc agagaggccg ggccacttcg ccggcatggt 1141 actcatttcg cctctggttc ttgccaatcc tgaatctgca acaactttca aggtccttgc 1201 tgcgaaagtg ctcaaccttg tgctgccaaa cttgtccctc gggcccatcg actccagcgt 1261 gctctctcgg aataagacag aggtcgacat ttataactca gaccccctga tctgccgggc 1321 agggctgaag gtgtgcttcg gcatccaact gctgaatgcc gtctcacggg tggagcgcgc 1381 cctccccaag ctgactgtgc ccttcctgct gctccagggc tctgccgatc gcctatgtga 1441 cagcaaaggg gcctacctgc tcatggagtt agccaagagc caggacaaga ctctcaagat 1501 ttatgaaggt gcctaccatg ttctccacaa ggagcttcct gaagtcacca actccgtctt 1561 ccatgaaata aacatgtggg tctctcaaag gacagccacg gcaggaactg cgtccccacc 1621 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga 1681 tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt 1741 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 1801 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 1861 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 1921 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 1981 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 2041 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 2101 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg 2161 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 2221 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 2281 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg 2341 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg 2401 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 2461 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 2521 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc 2581 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2641 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2701 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2761 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac 2821 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 2881 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 2941 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg 3001 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 3061 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3121 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3241 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3301 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 3361 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 3421 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 3481 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 3541 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 3601 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 3661 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 3721 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 3781 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 3841 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 3901 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 3961 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 4021 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 4081 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 4141 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 4201 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 4261 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 4321 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 4381 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 4441 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 4501 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 4561 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 4621 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 4681 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 4741 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 4801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 4861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 4921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 4981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 5041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 5101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 5161 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt 5221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 5281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 5341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 5401 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 5461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 5521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 5581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 5641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 5701 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 5761 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 5821 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 5881 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 5941 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 6001 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 6061 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 6121 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 6181 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 6241 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 6301 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 6361 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 6421 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 6481 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 6541 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 6601 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 6661 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 6721 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc 6781 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc 6841 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa 6901 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g //