LOCUS V043580 7680 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043580 VERSION V043580 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..2406 /label="FBXW4(NM_022039)" /note="FBXW4(NM_022039)" /gene="FBXW4" CDS 2413..2436 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" misc_feature 2456..3044 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3110..3158 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3360..3788 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3802..4131 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4198..4992 /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 5168..5289 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5338..5354) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5362..5378 /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(5386..5416) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5431..5452 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5740..6328) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6499..7359) /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(7360..7464) /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 ccaccatggg cagccagggc 721 cgctcggggc cccccgggaa cggcgggccc ggcgagggcg agggcggaga ggcgaggaag 781 ctgcaggaag ggagggtggc gagggggaag cgaaggaagg ggaaggggaa gggaaaagcg 841 agagcggggc aaggcggaag aggaagcggg gcggaaggga agcccgggcc gcagacggcg 901 aaggaggcag ccgggccggg ggctgacgcg ggagcgaggg catgcccaag ggaggaagca 961 gagggaggca gaagcgtgga ggaaggggcg agaggcatcg tcaagggagt cgaggggagc 1021 gcaggggccg ggaaggaggc acaaggaaga gagtatggga agaaggagga atggagggtc 1081 agggctaggc ggcgggaggg cgccaggccg ggaagagcac agggacgagg gggtcaggct 1141 tgggccgaca tcgcggggac aggggtggcc atggcggcgg cggccgggga ggaggaggag 1201 gaggaggagg cggctcggga gtcggctgcc cgcccggccg cggggcctgc gctctggcgc 1261 ctgccggagg agctgctgct gctcatctgc tcctacctgg acatgcgggc cctcggccgc 1321 ctggcccagg tgtgccgctg gctgcggcgc ttcaccagct gcgatctgct ctggcgccgg 1381 atagcccggg cctcgctcaa ctccggcttc acgcggctcg gcaccgacct gatgaccagt 1441 gtcccagtga aggaacgagt gaaggtgtct cagaactgga gactggggcg ctgccgagag 1501 gggattctgc tgaagtggag atgcagtcag atgccctgga tgcagctaga ggatgattct 1561 ctgtacatat cccaggctaa tttcatcctg gcctaccagt tccgtccaga tggtgccagc 1621 ttgaatcgtc ggcctctggg agtctttgct gggcatgatg aggacgtttg ccactttgtg 1681 ctggccaact cgcatattgt tagtgcagga ggggatggga agattggcat tcataagatt 1741 cacagcacct tcactgtcaa gtactcggct catgaacagg aggtgaactg tgtggattgc 1801 aaagggggca tcattgtgag tggctccagg gacaggacgg ccaaggtgtg gcctttggcc 1861 tcaggccggc tggggcagtg cttacacacc atccagactg aagaccgagt ctggtccatt 1921 gctatcagcc cattactcag ctcttttgtg acagggacgg cttgttgcgg gcacttctca 1981 cccctgagaa tctgggacct caacagtggg cagctgatga cacacttggg cagtgacttt 2041 cccccagggg ctggggtgct ggatgtcatg tatgagtccc ctttcacact gctgtcctgt 2101 ggctatgaca cctatgttcg ctactgggac ctccgcacca gcgtccggaa atgtgtcatg 2161 gagtgggagg agccccacga cagcaccctg tactgcctgc agacagatgg caaccacctg 2221 ctggccacag gttcctccta ctacggtgtt gtacggctgt gggaccggcg tcaaagggcc 2281 tgcctgcacg ccttcccgct gacgtcgact cccctcagca gccctgtgta ctgcctgcgt 2341 ctcaccacca agcatctcta tgctgccctg tcttacaacc tccacgtcct ggattttcaa 2401 aacccactcg agcaccatca ccatcaccat caccattagg gcccgatatc tcgacaatca 2461 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 2521 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 2581 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 2641 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 2701 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 2761 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 2821 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 2881 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 2941 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 3001 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg 3061 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa 3121 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca 3181 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc 3241 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt 3301 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca 3361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 3421 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 3481 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag 3541 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 3601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 3661 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 3721 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 3781 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 3841 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 3961 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 4021 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 4081 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 4141 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg 4201 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 4261 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 4321 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag 4381 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 4441 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 4501 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 4561 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 4621 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 4681 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc 4741 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 4801 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 4861 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 4921 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 4981 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct 5041 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 5101 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 5161 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 5221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 5281 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 5341 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 5401 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 5461 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 5521 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 5581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 5941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 6001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 6061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 6121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 6181 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 6241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 6301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 6361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 6421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 6481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 6541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 6601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 6661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 6721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 6781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 7021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 7081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 7141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 7201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 7261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 7321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 7381 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 7441 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg 7501 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca 7561 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc 7621 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg //