LOCUS V027509 7826 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027509 VERSION V027509 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..3316 /label="Nek4(NM_011849)" /note="Nek4(NM_011849)" /gene="Nek4" CDS 3323..3388 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3426..3650 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3696..4124 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4138..4467 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4534..5328 /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 5502..5623 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5672..5688) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5696..5712 /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(5720..5750) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5765..5786 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6074..6659) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6830..7690) /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(7691..7795) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgccccagg ccgcatactg 961 ctacatgcgg gtcgtgggca ggggtagtta cggggaggtg acgctcgtga agcatcggcg 1021 agacggcaag cagtatgtca tcaaaaagct gaacctccga aatgcctcca gccgagagcg 1081 tcgagctgct gagcaggaag ctcagctcct atctcagttg aagcacccca acattgtcac 1141 ctacaaagaa tcatgggagg gaggagacgg tctgctctac attgtcatgg gcttctgcga 1201 aggaggtgat ctgtaccgga agctcaagga gcagaaaggg cagcttctgc ctgagagtca 1261 ggtggtggag tggtttgtcc agatcgccat ggccttgcag tatttacatg agaaacacat 1321 ccttcatcga gatctgaaaa ctcagaatgt cttcttaaca agaacaaaca tcatcaaagt 1381 gggggaccta ggaattgccc gcgtcttaga gaatcacggt gacatggcga gcaccctcat 1441 tggcacaccc tattacatga gccctgagct gttctccaac aaaccctaca actataagtc 1501 ggatgtttgg gctttgggat gctgtgtata tgaaatggcc accctgaagc acgcttttaa 1561 tgcaaaagac atgaattctt tagtttatcg gattattgaa ggaaagttgc cgccaatgcc 1621 aaaagtttac agcacagagc tggctgagct gataagaaca atgctgagca gaaggcctga 1681 ggagaggccc tctgtgcgga gcatcctgag gcagccctac ataaaacacc acatctcctt 1741 gttcttggag gccaccaagg caaaaacttc caaaaataat gttaaaaact gtgactccag 1801 agccaagcct gttgctgctg tagtttccag aaaggaagaa tcaaatactg acgtgataca 1861 ctaccagcca cgctcttctg agggctctgc tttgcatgtc atgggtgaag ataaatgttt 1921 gtcccaagag aaaccagtgg acattggtcc cttgagatca ccggctagtc ttgaaggcca 1981 cactggcaaa caagacatga acaatactgg ggagtcatgt gccacaatca gtaggataaa 2041 tatcgatatc ttacctgcag aaaggaggga ctcagcgaac gctggtgtag ttcaggagag 2101 ccaaccacaa cacgtggatg ccgctgatga agtagacagt cagtgcagta tttctcaaga 2161 gaaggagagg ctacagggca ataccaagtc cagtgatcag cctggaaatc tccttcccag 2221 acggtcctct gatggtggtg atggggaagg gagtgaactg gtgaagccat tgtatccctc 2281 aaataaagac caaaagccag accaggatca agttactggt attatagaaa atcaggacag 2341 catccaccca agatcccagc cacacagctc tatgtctgaa ccttctttgt cccggcagcg 2401 aaggcagaaa aaaagggaac agactgccca tagtcgtaca aagagccagt tccaagaatt 2461 accccctcgg cttttgcctt cttatcctgg tattgggaaa gtggatatca tagcaacaca 2521 gcaaaatgat ggaaaccaag gtggacctgt ggctgggtgt gtaaacagtt caaggaccag 2581 ctcaacagca tcagcaaagg atcgaccatt atcagccaga gagaggagac ggctaaagca 2641 gtcacaggag gagatgctgc cctcaggccc tgcagttcag agaactccaa gtgcagttga 2701 gccactgaaa ccacaggaag aagaccaacc catccctgcc caacgattct cttctgactg 2761 cagcatcact cagatgaatc acacactccc gagggaaaag gagaagaggc tgatgcatgg 2821 cctgtctgag gatgagttaa gctcttctac aagctcaact gataagtcag atggggattc 2881 cagggagggg aagagtcata caaatgaaat gaaagacttg gtacagctga tgactcagac 2941 tctgaggctg gaagctaagg agagctgtga agatctccag gtcctaaacc cagggtcaga 3001 attcagactt catcgaaagt atcgagacac gctggtgctt catggcaagg ttgcagaaga 3061 ggtggaaccc cactgtacgg agctgcctac gggtatcata ccaggttctg aaaagatcag 3121 gagaatcgtt gaagtcttaa gagctgatgt aatccagggc ctggggattc aactcttaga 3181 gcaggtgttt gatcttttgg gagaggaaga tgagttggag agagaggcac gattacagga 3241 gcacatggga gacaagtata caacttactg tgtgaaagct cgccaattga aattttttga 3301 agaaaatgtg agttttctcg aggactacaa agaccatgac ggtgattata aagatcatga 3361 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 3421 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3481 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3541 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3601 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3661 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 3781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 3901 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 3961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4021 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 4081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 4141 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4201 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4261 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4321 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4381 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4441 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4501 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4561 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 4621 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4681 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 4741 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 4801 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 4861 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 4921 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 4981 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 5041 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 5101 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 5161 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 5221 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 5281 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 5341 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 5401 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5461 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5521 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5581 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5641 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5701 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 5761 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 5821 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 5881 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 5941 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 6001 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 6061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 6121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 6181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6241 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6481 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6601 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6661 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6721 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 6781 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 6841 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 6901 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 6961 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 7021 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 7081 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 7141 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 7201 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7261 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7321 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7381 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7441 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7501 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7561 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 7621 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 7681 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 7741 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 7801 ttccccgaaa agtgccacct gacgtc //