LOCUS V025373 6472 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025373 VERSION V025373 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 931..1965 /label="Aurkb(NM_011496)" /note="Aurkb(NM_011496)" /gene="Aurkb" CDS 1975..1992 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 2039..2263 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2309..2737 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2751..3080 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3147..3941 /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 4115..4236 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4285..4301) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4309..4325 /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(4333..4363) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4378..4399 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4687..5272) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5443..6303) /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(6304..6408) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cactagtcca gtgtggtgga attcgccacc atggctcaga aggagaacgc ctacccgtgg 961 ccctacggct caaagacgtc tcagtctggc ctgaacacgt tgtcccagag agtcctacgg 1021 aaggagcccg ccacgacatc tgcgcttgct ctcgtgaacc ggttcaacag ccagtccaca 1081 gctgcccctg gccagaagtt ggctgagaac aagagtcagg gctccactgc ctcgcaagga 1141 tcccagaaca agcagccttt cactattgac aactttgaga ttgggcgtcc tttgggcaaa 1201 ggcaaatttg gaaacgtgta cttggctcgg gagaagaaga gccgtttcat cgtggcactc 1261 aagatcctct tcaagtctca gattgagaag gagggggtag agcaccagct tcgccgagag 1321 atcgaaatcc aggcgcacct gaaacatccc aacatccttc aactctacaa ctacttctac 1381 gaccagcaga ggatctactt aatcctggaa tacgcccctc gcggggaact ctacaaggaa 1441 ctgcagaaga gtcggacctt cgatgagcag cggactgcca cgatcatgga ggaactgtca 1501 gatgccctga cctactgcca caagaagaag gtaattcaca gagacataaa gccggagaac 1561 ctgctgttag gtctgcaggg agaactgaag attgcagact ttggctggtc ggtgcatgcc 1621 ccatccctga ggaggaagac catgtgcggc acgctggact atctgccccc agagatgatt 1681 gaggggcgca tgcataatga aatggtagat ctatggtgca tcggggtgct ctgctatgaa 1741 ctgatggtgg ggaacccacc cttcgagagc cctagccaca gtgagacgta tcgtcggatt 1801 gtcaaggtgg acctgaagtt cccctcttct gtgccttcgg gcgcccagga cctcatctcc 1861 aagctgctca aacataaccc ctggcaacgg ctgcccctgg cggaggttgc agctcaccct 1921 tgggtccggg ccaactcaag gagggttctg cctccctctg cccttgcggc cgcacatcac 1981 catcatcacc attgactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact 2041 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 2101 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 2161 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 2221 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 2281 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 2341 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 2461 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 2521 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 2641 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 2701 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 2761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 2821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 2941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3001 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 3061 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 3121 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 3181 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 3241 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 3301 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 3361 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 3421 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 3481 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 3541 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 3601 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 3661 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 3721 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 3781 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 3841 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 3901 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 3961 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 4021 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 4081 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 4141 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 4201 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 4261 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 4321 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 4381 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 4441 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 4501 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 4561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 4621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 4681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 4741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 4801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 4861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 4921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 4981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 5041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 5101 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 5161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 5221 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 5281 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 5341 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5401 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 5461 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5521 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5581 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5641 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5701 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5761 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5821 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5881 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 5941 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 6001 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6061 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 6121 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 6181 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 6241 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 6301 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 6361 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 6421 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gt //