LOCUS V040439 7450 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040439 VERSION V040439 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 706..2106 /label="MAP3K8(NM_005204)" /note="MAP3K8(NM_005204)" /gene="MAP3K8" CDS 2113..2202 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 2226..2814 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2880..2928 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3130..3558 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3572..3901 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3968..4762 /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 4938..5059 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5108..5124) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5132..5148 /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(5156..5186) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5201..5222 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5510..6098) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6269..7129) /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(7130..7234) /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgga gtacatgagc 721 actggaagtg acaataaaga agagattgat ttattaatta aacatttaaa tgtgtctgat 781 gtaatagaca ttatggaaaa tctttatgca agtgaagagc cagcagttta tgaacccagt 841 ctaatgacca tgtgtcaaga cagtaatcaa aacgatgagc gttctaagtc tctgctgctt 901 agtggccaag aggtaccatg gttgtcatca gtcagatatg gaactgtgga ggatttgctt 961 gcttttgcaa accatatatc caacactgca aagcattttt atggacaacg accacaggaa 1021 tctggaattt tattaaacat ggtcatcact ccccaaaatg gacgttacca aatagattcc 1081 gatgttctcc tgatcccctg gaagctgact tacaggaata ttggttctga ttttattcct 1141 cggggcgcct ttggaaaggt atacttggca caagatataa agacgaagaa aagaatggcg 1201 tgtaaactga tcccagtaga tcaatttaag ccatctgatg tggaaatcca ggcttgcttc 1261 cggcacgaga acatcgcaga gctgtatggc gcagtcctgt ggggtgaaac tgtccatctc 1321 tttatggaag caggcgaggg agggtctgtt ctggagaaac tggagagctg tggaccaatg 1381 agagaatttg aaattatttg ggtgacaaag catgttctca agggacttga ttttctacac 1441 tcaaagaaag tgatccatca tgatattaaa cctagcaaca ttgttttcat gtccacaaaa 1501 gctgttttgg tggattttgg cctaagtgtt caaatgaccg aagatgtcta ttttcctaag 1561 gacctccgag gaacagagat ttacatgagc ccagaggtca tcctgtgcag gggccattca 1621 accaaagcag acatctacag cctgggggcc acgctcatcc acatgcagac gggcacccca 1681 ccctgggtga agcgctaccc tcgctcagcc tatccctcct acctgtacat aatccacaag 1741 caagcacctc cactggaaga cattgcagat gactgcagtc cagggatgag agagctgata 1801 gaagcttccc tggagagaaa ccccaatcac cgcccaagag ccgcagacct actaaaacat 1861 gaggccctga acccgcccag agaggatcag ccacgctgtc agagtctgga ctctgccctc 1921 ttggagcgca agaggctgct gagtaggaag gagctggaac ttcctgagaa cattgctgat 1981 tcttcgtgca caggaagcac cgaggaatct gagatgctca agaggcaacg ctctctctac 2041 atcgacctcg gcgctctggc tggctacttc aatcttgttc ggggaccacc aacgcttgaa 2101 tatggcctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt 2161 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc 2221 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 2281 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 2341 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 2401 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 2461 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 2521 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 2581 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 2641 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 2701 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 2761 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa 2821 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac 2881 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg 2941 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 3001 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 3061 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg 3121 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 3181 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 3241 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt 3301 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 3361 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 3421 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 3481 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac 3541 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 3721 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 3781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 3841 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 3901 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc 3961 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 4021 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 4081 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 4141 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 4201 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 4261 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 4321 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 4381 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 4441 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 4501 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 4561 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 4621 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 4681 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 4741 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga 4801 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 4861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 4921 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4981 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5041 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 5101 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 5341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 5401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 5461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 5521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 5581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 5641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 5701 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 5761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 5821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 5881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 5941 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 6061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 6481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 6541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 6961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7261 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc 7321 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta 7381 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa 7441 tctgcttagg //