LOCUS V020639 7202 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020639 VERSION V020639 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 908..2695 /label="MYLK2(NM_033118)" /note="MYLK2(NM_033118)" /gene="MYLK2" CDS 2705..2722 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 2769..2993 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3039..3467 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3481..3810 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3877..4671 /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 4845..4966 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5015..5031) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5039..5055 /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(5063..5093) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5108..5129 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5417..6002) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6173..7033) /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(7034..7138) /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 cgccaccatg gcgacagaaa atggagcagt tgagctggga attcagaacc catcaacaga 961 caaggcacct aaaggtccca caggtgaaag acccctggct gcagggaaag accctggccc 1021 cccagaccca aagaaagctc cggatccacc caccctgaag aaagatgcca aagcccctgc 1081 ctcagagaaa ggggatggta ccctggccca accctcaact agcagccaag gccccaaagg 1141 agagggtgac aggggcgggg ggcccgcgga gggcagtgct gggcccccgg cagccctgcc 1201 ccagcagact gcgacacctg agaccagcgt caagaagccc aaggctgagc agggagcctc 1261 aggcagccag gatcctggaa agcccagggt gggcaagaag gcagcagagg gccaagcagc 1321 agccaggagg ggctcacctg cctttctgca tagccccagc tgtcctgcca tcatctccag 1381 ttctgagaag ctgctggcca agaagccccc aagcgaggca tcagagctca cctttgaagg 1441 ggtgcccatg acccacagcc ccacggatcc caggccagcc aaggcagaag aaggaaagaa 1501 catcctggca gagagccaga aggaagtggg agagaaaacc ccaggccagg ctggccaggc 1561 taagatgcaa ggggacacct cgagggggat tgagttccag gctgttccct cagagaaatc 1621 cgaggtgggg caggccctct gtctcacagc cagggaggag gactgcttcc agattttgga 1681 tgattgcccg ccacctccgg cccccttccc tcaccgcatg gtggagctga ggaccgggaa 1741 tgtcagcagt gaattcagta tgaactccaa ggaggcgctc ggaggtggca agtttggggc 1801 agtctgtacc tgcatggaga aagccacagg cctcaagctg gcagccaagg tcatcaagaa 1861 acagactccc aaagacaagg aaatggtgtt gctggagatt gaggtcatga accagctgaa 1921 ccaccgcaat ctgatccagc tgtatgcagc catcgagact ccgcatgaga tcgtcctgtt 1981 catggagtac atcgagggcg gagagctctt cgagaggatt gtggatgagg actaccatct 2041 gaccgaggtg gacaccatgg tgtttgtcag gcagatctgt gacgggatcc tcttcatgca 2101 caagatgagg gttttgcacc tggacctcaa gccagagaac atcctgtgtg tcaacaccac 2161 cgggcatttg gtgaagatca ttgactttgg cctggcacgg aggtataacc ccaacgagaa 2221 gctgaaggtg aactttggga ccccagagtt cctgtcacct gaggtggtga attatgacca 2281 aatctccgat aagacagaca tgtggagtat gggggtgatc acctacatgc tgctgagcgg 2341 cctctccccc ttcctgggag atgatgacac agagacccta aacaacgttc tatctggcaa 2401 ctggtacttt gatgaagaga cctttgaggc cgtatcagac gaggccaaag actttgtctc 2461 caacctcatc gtcaaggacc agagggcccg gatgaacgct gcccagtgtc tcgcccatcc 2521 ctggctcaac aacctggcgg agaaagccaa acgctgtaac cgacgcctta agtcccagat 2581 cttgcttaag aaatacctca tgaagaggcg ctggaagaaa aacttcattg ctgtcagcgc 2641 tgccaaccgc ttcaagaaga tcagcagctc gggggcactg atggctctgg gggtcgcggc 2701 cgcacatcac catcatcacc attgactcga gtctagaggg cccgtttaaa cccgctgatc 2761 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 2821 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 2881 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 2941 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 3001 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 3061 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 3121 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 3181 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 3241 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 3301 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 3361 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 3421 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 3481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3541 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 3601 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 3661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 3721 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 3781 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 3841 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3901 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3961 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 4021 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 4081 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4141 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4201 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 4261 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4321 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 4381 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 4441 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 4501 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 4561 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 4621 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4681 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4741 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4801 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 4861 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 4921 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 4981 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 5041 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 5101 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 5161 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 5221 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 5281 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 5341 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 5401 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 5461 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 5521 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 5581 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 5641 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 5701 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 5761 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 5821 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 5881 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 5941 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 6001 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6061 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 6121 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 6181 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 6241 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 6301 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 6361 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 6421 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6481 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6781 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 7021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 7141 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg 7201 gt //