LOCUS V032167 8530 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032167 VERSION V032167 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..3186 /label="Map4k1(NM_008279)" /note="Map4k1(NM_008279)" /gene="Map4k1" CDS 3193..3282 /codon_start=1 /product="3 tandem Myc epitope tags" /transl_table=1 /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" /label="3xMyc" misc_feature 3306..3894 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3960..4008 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4210..4638 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4652..4981 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5048..5842 /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 6018..6139 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6188..6204) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6212..6228 /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(6236..6266) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6281..6302 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6590..7178) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7349..8209) /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(8210..8314) /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 ccaccatggc ccttgtggac 721 cccgacattt ttaataagga ccctcgagaa cactatgatc tactgcaacg actgggtggt 781 ggaacttatg gagaagtatt caaggctcgc gacaaggtgt ctaaggacct ggtggctttg 841 aagatggtga agatggagcc tgatgatgac gtcgccaccc ttcagaagga gattctcatg 901 ctgaaaactt gccggcacgc caacatcgtg gcttaccatg gcagttacct ctggctgcag 961 aagctgtgga tctgcatgga attctgtgga gctggttctc tccaggacat ctaccaagtg 1021 actggctctc tgtccgagct ccagatcagc tatgtgtgcc gggaagtgct tcagggactg 1081 gcctacctgc attcagagaa gaagatacat cgagacatca agggagccaa catcctcatc 1141 aacgactgcg gggaggttaa gctggctgac tttgggatct cagcccagat tggggcgaca 1201 ctggcccggc gcctttcttt cattgggaca ccgtactgga tggctccaga ggtggcagcg 1261 gtggccctga agggtggata caatgagctg tgcgacatct ggtctcttgg catcacagcc 1321 atcgagctgg ctgagcttca gccccctctc tttgatgtgc accctctcag ggttctcttc 1381 ctcatgacca agagcgggta ccagcctcct cggctgaagg aaaagagcag atggtcatct 1441 tccttccaca actttgtcaa agtcaccctc actaagaact ccaagaaacg ccccagtgcc 1501 accaagatgc tcagtcatca gctggtgtcc cagccagggc tgaacagagg cctgatcttg 1561 gatcttcttg acaaaatgaa gaatcctggg aaggggctgc ctgttgatat tgaagatgag 1621 gagcctgagc caccccctgc catccctcgg cggatcagat ctacctaccg ggccagctca 1681 ctagggatcc cagatgcaga ctgctgccgg aggcaaatgg agttccagag gcccagatgc 1741 gtggactgca gacctcaggc tgaaactgtt cgcttatatc cccctgcaca cttcgggagc 1801 accagtccca ggagtcagct gtcagattct gatgacgact atgatgatgt ggacatccct 1861 gccccttcag agaacatacc tcctccactg ccccccaagc ccaagtttcg gtctccatca 1921 gatgatggtt ctggaggaat tagggacgat gggcaattga gcccaggggt tctggttcga 1981 tgtgcaagtg ggccccctcc acgtactccc cgacctggac ctcccccagc gacctgcagc 2041 cctcacctca cagctcgctc agatccctca ttgtggaacc cagctgctcc ggagccaggc 2101 cagcccccac tcgtaccccc aaggaaggaa aagatgaggg gaaagatgga gaacgagaag 2161 cgcagagaga aatatcccct tcttgtgaaa ttattcaatg gctgtcccct tcagatccac 2221 agcacagctg cctggacaca tccctctaca aaagatcaga acctgctcct gggggcagag 2281 gaaggcattt tcatactgaa ccggaatgat caggaggcca cgctggaaat gatcttccct 2341 ggccgcacaa cctggctcta ttgcatcaac aacctcctca tgtctctctc agggaagaca 2401 ccctacctgt attctcatag catcctgggc ctgcttgaaa ggaaagacgg cagaacggga 2461 agccccattg ctcacattag ccctcaccgg ttactggcaa gaaagaacat ggtctccagt 2521 aagatccagg acaccaaagg ctgccgggct tgctgtgtgg ctgagagtgc tagctctggg 2581 ggcccattct tatgtggggc attggagacg tctgtggttt tgctccagtg gtaccaacca 2641 atgaacaaat tcttgcttgt ccggcaggtg ctgttcccgc tgcccacgcc gctgccggtg 2701 ttcacgctgc tgacaacgcc gggctccgag ctgccagctg tgtgcatcgg cgtgagcccg 2761 gggcaggcgg caaagtcggt gctcttccac accgtgcgct tcggtgcgct gtcctgctgg 2821 ctggacgact ccagcacgga gcacaaggga ccagtgcagg tgatccaagt aaaggaagac 2881 atggtgatgg tgttgatgga tggctctcta aagctagtga ccccagaggg ggcaccagct 2941 cctgggctcc gcacccctga gatccccatg acagaagcgg tcgaggctgt ggctatggtt 3001 gaagatcgac tcgaggcctt ctggaagcat ggagttcagg tgtgggctcc aggcttgaaa 3061 cagccgctgc aggagctgag agaccccacc ctcaccttcc gtctgctctg ctcccccagg 3121 ccggtagtgg tggagacaag gcccacggat gaccctacgg cccccagcaa cctctacatc 3181 caggaactcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt 3241 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc 3301 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 3361 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 3421 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 3481 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 3541 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 3601 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 3661 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 3721 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 3781 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 3841 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa 3901 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac 3961 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg 4021 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 4081 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 4141 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg 4201 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 4261 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 4321 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt 4381 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 4441 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 4501 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 4561 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac 4621 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 4681 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 4741 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 4801 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 4861 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 4921 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 4981 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc 5041 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 5101 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 5161 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 5221 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 5281 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 5341 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 5401 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 5461 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 5521 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 5581 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 5641 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 5701 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 5761 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 5821 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga 5881 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 5941 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 6001 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 6061 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 6121 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 6181 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 6241 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 6301 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 6361 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 6421 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 6481 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 6541 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 6601 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 6661 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 6721 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 6781 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 6841 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 6901 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 6961 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 7021 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 7081 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 7141 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 7201 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 7261 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 7321 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 7381 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 7441 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 7501 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 7561 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 7621 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 7681 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 7741 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 7801 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 7861 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 7921 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 7981 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 8041 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 8101 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 8161 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 8221 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 8281 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 8341 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc 8401 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta 8461 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa 8521 tctgcttagg //