LOCUS V019654 9356 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019654 VERSION V019654 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..4036 /label="MAP3K9(NM_033141)" /note="MAP3K9(NM_033141)" /gene="MAP3K9" CDS 4043..4108 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 4132..4720 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4786..4834 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5036..5464 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5478..5807 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5874..6668 /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 6844..6965 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(7014..7030) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7038..7054 /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(7062..7092) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7107..7128 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7416..8004) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8175..9035) /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(9036..9140) /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 ccatggagcc ctccagagcg cttctcggct gcctagcgag 721 cgccgccgct gccgccccgc cgggggagga tggagcaggg gccggggccg aggaggagga 781 ggaggaggag gaggaggcgg cggcggcggt gggccccggg gagctgggct gcgacgcgcc 841 gctgccctac tggacggccg tgttcgagta cgaggcggcg ggcgaggacg agctgaccct 901 gcggctgggc gacgtggtgg aggtgctgtc caaggactcg caggtgtccg gcgacgaggg 961 ctggtggacc gggcagctga accagcgggt gggcatcttc cccagcaact acgtgacccc 1021 gcgcagcgcc ttctccagcc gctgccagcc cggcggcgag gaccccagtt gctacccgcc 1081 cattcagttg ttagaaattg attttgcgga gctcaccttg gaagagatta ttggcatcgg 1141 gggctttggg aaggtctatc gtgctttctg gataggggat gaggttgctg tgaaagcagc 1201 tcgccacgac cctgatgagg acatcagcca gaccatagag aatgttcgcc aagaggccaa 1261 gctcttcgcc atgctgaagc accccaacat cattgcccta agaggggtat gtctgaagga 1321 gcccaacctc tgcttggtca tggagtttgc tcgtggagga cctttgaata gagtgttatc 1381 tgggaaaagg attcccccag acatcctggt gaattgggct gtgcagattg ccagagggat 1441 gaactactta catgatgagg caattgttcc catcatccac cgcgacctta agtccagcaa 1501 catattgatc ctccagaagg tggagaatgg agacctgagc aacaagattc tgaagatcac 1561 tgattttggc ctggctcggg aatggcaccg aaccaccaag atgagtgcgg cagggacgta 1621 tgcttggatg gcacccgaag tcatccgggc ctccatgttt tccaaaggca gtgatgtgtg 1681 gagctatggg gtgctacttt gggagttgct gactggtgag gtgccctttc gaggcattga 1741 tggcttagca gtcgcttatg gagtggccat gaacaaactc gcccttccta ttccttctac 1801 gtgcccagaa ccttttgcca aactcatgga agactgctgg aatcctgatc cccactcacg 1861 accatctttc acgaatatcc tggaccagct aaccaccata gaggagtctg gtttctttga 1921 aatgcccaag gactccttcc actgcctgca ggacaactgg aaacacgaga ttcaggagat 1981 gtttgaccaa ctcagggcca aagaaaagga acttcgcacc tgggaggagg agctgacgcg 2041 ggctgcactg cagcagaaga accaggagga actgctgcgg cgtcgggagc aggagctggc 2101 cgagcgggag attgacatcc tggaacggga gctcaacatc atcatccacc agctgtgcca 2161 ggagaagccc cgggtgaaga aacgcaaggg caagttcagg aagagccggc tgaagctcaa 2221 ggatggcaac cgcatcagcc tcccttctga tttccagcac aagttcacgg tgcaggcctc 2281 ccctaccatg gataaaagga agagtcttat caacagccgc tccagtcctc ctgcaagccc 2341 caccatcatt cctcgccttc gagccatcca gttgacacca ggtgaaagca gcaaaacctg 2401 gggcaggagc tcagtcgtcc caaaggagga aggggaggag gaggagaaga gggccccaaa 2461 gaagaaggga cggacgtggg ggccagggac gcttggtcag aaggagcttg cctcgggaga 2521 tgaaggatcc cctcagagac gtgagaaagc taatggttta agtaccccat cagaatctcc 2581 acatttccac ttgggcctca agtccctggt agatggatat aagcagtggt cgtccagtgc 2641 ccccaacctg gtgaagggcc caaggagtag cccggccctg ccagggttca ccagccttat 2701 ggagatggcc ttgctggcag ccagttgggt ggtgcccatc gacattgaag aggatgagga 2761 cagtgaaggc ccagggagtg gagagagtcg cctacagcat tcacccagcc agtcctacct 2821 ctgtatccca ttccctcgtg gagaggatgg cgatggcccc tccagtgatg gaatccatga 2881 ggagcccacc ccagtcaact cggccacgag tacccctcag ctgacgccaa ccaacagcct 2941 caagcggggc ggtgcccacc accgccgctg cgaggtggct ctgctcggct gtggggctgt 3001 tctggcagcc acaggcctag ggtttgactt gctggaagct ggcaagtgcc agctgcttcc 3061 cctggaggag cctgagccac cagcccggga ggagaagaaa agacgggagg gtctttttca 3121 gaggtccagc cgtcctcgtc ggagcaccag ccccccatcc cgaaagcttt tcaagaagga 3181 ggagcccatg ctgttgctag gagacccctc tgcctccctg acgctgctct ccctctcctc 3241 catctccgag tgcaactcca cacgctccct gctgcgctcc gacagcgatg aaattgtcgt 3301 gtatgagatg ccagtcagcc cagtcgaggc ccctcccctg agtccatgta cccacaaccc 3361 cctggtcaat gtccgagtag agcgcttcaa acgagatcct aaccaatctc tgactcccac 3421 ccatgtcacc ctcaccaccc cctcgcagcc cagcagtcac cggcggactc cttctgatgg 3481 ggcccttaag ccagagactc tcctagccag caggagcccc tccagcaatg ggttgagccc 3541 cagtcctgga gcaggaatgt tgaaaacccc cagtcccagc cgagacccag gtgaattccc 3601 ccgtctccct gaccccaatg tggtcttccc cccaacccca aggcgctgga acactcagca 3661 ggactctacc ttggagagac ccaagactct ggagtttctg cctcggccgc gtccttctgc 3721 caaccggcaa cggctggacc cttggtggtt tgtgtccccc agccatgccc gcagcacctc 3781 cccagccaac agctccagca cagagacgcc cagcaacctg gactcctgct ttgctagcag 3841 tagcagcact gtagaggagc ggcctggact tccagccctg ctcccgttcc aggcagggcc 3901 gctgcccccg actgagcgga cgctcctgga cctggatgca gaggggcaga gtcaggacag 3961 caccgtgccg ctgtgcagag cggaactgaa cacacacagg cctgcccctt atgagatcca 4021 gcaggagttc tggtctctcg aggactacaa agaccatgac ggtgattata aagatcatga 4081 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct 4141 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 4201 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 4261 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 4321 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 4381 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 4441 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 4501 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 4561 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 4621 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 4681 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa 4741 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 4801 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 4861 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 4921 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 4981 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg 5041 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 5101 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 5161 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct 5221 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 5281 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 5341 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 5401 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 5461 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 5521 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 5581 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 5641 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 5701 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 5761 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 5821 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 5881 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 5941 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 6001 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 6061 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 6121 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 6181 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 6241 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 6301 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 6361 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 6421 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 6481 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 6541 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 6601 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 6661 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca 6721 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 6781 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 6841 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6901 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6961 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 7021 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 7081 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 7141 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 7201 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 7261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 7321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 7381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 7441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 7501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 7561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 7621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 7681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 7741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 7801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 7861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 7921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 7981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 8041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 8101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 8161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 8221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 8281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 8341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 8401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 8461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 8521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 8581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 8641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 8701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 8761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 8821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 8881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 8941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 9001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 9061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 9121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 9181 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt 9241 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 9301 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg //