LOCUS V022062 7517 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022062 VERSION V022062 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 677..2170 /label="NMT2(NM_004808)" /note="NMT2(NM_004808)" /gene="NMT2" CDS 2204..2269 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2293..2881 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2947..2995 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3197..3625 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3639..3968 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4035..4829 /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 5005..5126 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5175..5191) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5199..5215 /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(5223..5253) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5268..5289 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5577..6165) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6336..7196) /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(7197..7301) /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 gccaccatgg cggaggacag cgagtctgcg gccagccagc agagcctgga 721 actggacgac caggacacgt gcgggataga cggggacaat gaggaggaga cggagcacgc 781 caaaggaagt cctggagggt atttgggagc caaaaagaaa aagaagaaac agaagagaaa 841 aaaggagaaa ccaaattccg gaggcaccaa gtcagactcg gcatctgatt cccaggagat 901 taaaattcag cagccttcga aaaatcccag tgttccaatg cagaagttgc aggatatcca 961 gagagcaatg gagctgctat ccgcatgcca gggcccagcc aggaacattg atgaggctgc 1021 aaagcacaga taccagtttt gggacacaca accggtacca aaactagatg aagtcataac 1081 atctcatggt gcaattgaac cagataaaga caacgtacgc caagaaccgt attctttgcc 1141 acagggtttt atgtgggaca ctttagactt gagtgatgcc gaagtgctca aggagttata 1201 cacgttgtta aatgagaatt acgtagaaga tgatgacaat atgttccgat ttgactattc 1261 acccgagttc ctgttgtggg ctctgcgtcc accaggctgg ctcctgcagt ggcactgtgg 1321 ggtcagagtg tcttcaaata aaaaactggt cgggttcata agtgccatcc cagcaaacat 1381 tcggatttat gacagtgtga agaagatggt agaaatcaac tttctttgtg ttcataagaa 1441 gttgagatcg aaacgggtag ccccagtgct aatccgagag atcactagaa gagtgaacct 1501 ggaagggatc ttccaggctg tgtacaccgc gggagtggtt cttcctaagc ccatagccac 1561 atgcagatac tggcatcgat cactaaaccc cagaaaattg gtagaagtga aattttctca 1621 cttgagtaga aatatgactt tacagagaac aatgaagcta tacagacttc cagatgttac 1681 aaagacttca ggtttgagac caatggaacc aaaagatatc aaatcagttc gagaattaat 1741 caacacttac ctgaagcagt ttcatctggc tccagtgatg gatgaagagg aagtagccca 1801 ctggttcctc ccccgggagc acattattga cacgtttgta gtggagagcc ccaacggtaa 1861 actgactgat ttcctgagct tctatacgct cccctccacg gtgatgcacc accctgctca 1921 caagagcctc aaagccgcct actcattcta caacatccac acagagacgc ccctgctgga 1981 cctcatgagc gacgcgctca tcctggctaa atcgaaagga tttgatgtat tcaatgcact 2041 ggatttgatg gaaaataaga cattcttgga aaaactcaag tttggtatag gagatggcaa 2101 tttgcagtat tacctgtaca attggaggtg tccaggtaca gattctgaaa aggttggact 2161 agtactacaa tgcagatatc cagcacagtg gcggccgctc gaggactaca aagaccatga 2221 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2281 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 2341 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 2401 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 2461 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 2521 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 2581 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 2641 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt 2701 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 2761 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 2821 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 2881 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg 2941 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa 3001 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 3061 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 3121 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc 3181 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3241 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3301 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct 3361 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3421 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3481 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3541 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg 3601 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3721 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3781 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 3841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 3901 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 3961 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 4021 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 4081 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 4141 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 4201 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 4261 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 4321 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4381 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4441 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4501 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4561 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4621 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4681 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4741 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4801 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac 4861 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 4921 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 4981 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 5041 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 5101 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 5161 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 5221 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 5281 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 5341 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 5401 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 5461 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 5521 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 5581 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5641 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5701 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 5761 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 5821 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 5881 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 5941 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 6001 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 6061 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 6121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 6181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 6241 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 6301 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 6361 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 6421 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 6481 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 6541 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 6601 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 6661 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 6721 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 6781 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 6841 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 6901 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 6961 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 7021 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 7081 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 7141 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 7201 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 7261 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 7321 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa 7381 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg 7441 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca 7501 tgaagaatct gcttagg //