LOCUS V033719 6974 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033719 VERSION V033719 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 141..157 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 318..697 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 698..901 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 935..958 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 962..2734 /label="PAK4(NM_005884)" /note="PAK4(NM_005884)" /gene="PAK4" CDS 2786..3304 /codon_start=1 /product="N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNG IKANFKIRHNIE" /label="VN173" polyA_signal 3315..3937 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" promoter 3966..4295 /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(4334..4352) /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" primer_bind complement(4366..4382) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4390..4406 /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(4414..4444) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4459..4480 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4768..5356) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5527..6387) /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(6388..6492) /gene="bla" /label="AmpR promoter" rep_origin 6519..6947 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct 961 tatgtttggg aagaggaaga agcgggtgga gatctccgcg ccgtccaact tcgagcaccg 1021 cgtgcacacg ggcttcgacc agcacgagca gaagttcacg gggctgcccc gccagtggca 1081 gagcctgatc gaggagtcgg ctcgccggcc caagcccctc gtcgaccccg cctgcatcac 1141 ctccatccag cccggggccc ccaagaccat cgtgcggggc agcaaaggtg ccaaagatgg 1201 ggccctcacg ctgctgctgg acgagtttga gaacatgtcg gtgacacgct ccaactccct 1261 gcggagagac agcccgccgc cgcccgcccg tgcccgccag gaaaatggga tgccagagga 1321 gccggccacc acggccagag ggggcccagg gaaggcaggc agccgaggcc ggttcgccgg 1381 tcacagcgag gcgggtggcg gcagtggtga caggcgacgg gcggggccag agaagaggcc 1441 caagtcttcc agggagggct cagggggtcc ccaggagtcc tcccgggaca aacgccccct 1501 ctccgggcct gatgtcggca ccccccagcc tgctggtctg gccagtgggg cgaaactggc 1561 agctggccgg ccctttaaca cctacccgag ggctgacacg gaccacccat cccggggtgc 1621 ccagggggag cctcatgacg tggcccctaa cgggccatca gcggggggcc tggccatccc 1681 ccagtcctcc tcctcctcct cccggcctcc cacccgagcc cgaggtgccc ccagccctgg 1741 agtgctggga ccccacgcct cagagcccca gctggcccct ccagcctgca cccccgccgc 1801 ccctgctgtt cctgggcccc ctggcccccg ctcaccacag cgggagccac agcgagtatc 1861 ccatgagcag ttccgggctg ccctgcagct ggtggtggac ccaggcgacc cccgctccta 1921 cctggacaac ttcatcaaga ttggcgaggg ctccacgggc atcgtgtgca tcgccaccgt 1981 gcgcagctcg ggcaagctgg tggccgtcaa gaagatggac ctgcgcaagc agcagaggcg 2041 cgagctgctc ttcaacgagg tggtaatcat gagggactac cagcacgaga atgtggtgga 2101 gatgtacaac agctacctgg tgggggacga gctctgggtg gtcatggagt tcctggaagg 2161 aggcgccctc accgacatcg tcacccacac caggatgaac gaggagcaga tcgcggccgt 2221 gtgccttgca gtgctgcagg ccctgtcggt gctccacgcc cagggcgtca tccaccggga 2281 catcaagagc gactcgatcc tgctgaccca tgatggcagg gtgaagctgt cagactttgg 2341 gttctgcgcc caggtgagca aggaagtgcc ccgaaggaag tcgctggtcg gcacgcccta 2401 ctggatggcc ccagagctca tctcccgcct tccctacggg ccagaggtag acatctggtc 2461 gctggggata atggtgattg agatggtgga cggagagccc ccctacttca acgagccacc 2521 cctcaaagcc atgaagatga ttcgggacaa cctgccaccc cgactgaaga acctgcacaa 2581 ggtgtcgcca tccctgaagg gcttcctgga ccgcctgctg gtgcgagacc ctgcccagcg 2641 ggccacggca gccgagctgc tgaagcaccc attcctggcc aaggcagggc cgcctgccag 2701 catcgtgccc ctcatgcgcc agaaccgcac cagaggaggt ggaggttcag gtggtggagg 2761 atcttctaga agatccatcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 2821 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 2881 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg 2941 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt 3001 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 3061 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 3121 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 3181 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 3241 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat 3301 cgagtaggga tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg 3361 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt 3421 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc 3481 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag 3541 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc 3601 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt 3661 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg 3721 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct 3781 tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag acacagcata 3841 ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct 3901 catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt 3961 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 4021 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 4141 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 4201 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 4261 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc 4321 agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc 4381 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 4441 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 4501 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 4561 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 4621 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 4681 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 4741 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 4801 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 4861 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 4921 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 4981 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 5041 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 5101 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 5161 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 5221 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 5281 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 5341 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5401 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 5461 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5521 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5581 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 5641 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 5701 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 5761 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5821 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 5881 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 5941 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6001 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6061 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6121 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6181 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 6241 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6301 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6361 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6421 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6481 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt 6541 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 6601 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 6661 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 6721 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 6781 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 6841 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 6901 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt 6961 aacgcttaca attt //