LOCUS V030961 6114 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030961 VERSION V030961 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1029..2261 /label="Kcnk2(NM_010607)" /note="Kcnk2(NM_010607)" /gene="Kcnk2" CDS 2280..2309 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2328..2351 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(2371..2387) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 2398..3020 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3169..3757) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 4086..4133 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(4365..5159) /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" promoter complement(5194..5551) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(5553..5657) /gene="bla" /label="AmpR promoter" rep_origin 5684..6112 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccat ggcggcccct gacttgctgg atcccaagtc tgctgctcag aactccaaac 1081 cgaggctctc attctcctca aaacccaccg tgcttgcttc ccgggtggag agtgactcgg 1141 ccattaatgt tatgaaatgg aagacagtct ccacgatttt cctggtggtc gtcctctacc 1201 tgatcatcgg agccacggtg ttcaaggcat tggagcagcc tcaggagatt tcccagagga 1261 ccaccattgt gatccagaag cagaccttca tagcccagca tgcctgcgtc aactccaccg 1321 agctggacga actcatccag caaatagtgg cagcaataaa cgcagggatt atccccttag 1381 gaaacagctc caatcaagtt agtcactggg acctcggaag ctctttcttc tttgctggta 1441 ctgttatcac aaccatagga tttggaaaca tctccccacg aactgaaggt ggaaaaatat 1501 tctgcatcat ctatgccttg ctgggaattc ccctctttgg ctttctactg gctggggttg 1561 gtgatcagct aggaactata tttggaaaag gaattgccaa agtggaagac acatttatta 1621 agtggaatgt tagtcagacg aagattcgta tcatctccac catcatcttc atcctgtttg 1681 gctgtgtcct ctttgtggct ctccctgcgg tcatattcaa gcacatagaa ggctggagcg 1741 ccctggacgc tatctatttt gtggttatca ctctgacgac cattggattt ggagactacg 1801 tggcaggtgg atcagacatt gaatatctgg acttctacaa gcctgtggtg tggttctgga 1861 tcctcgttgg gctggcctac tttgcagctg ttctgagcat gattggggac tggctacggg 1921 tgatctctaa gaagacgaag gaagaggtgg gagagttcag agcgcatgcc gctgagtgga 1981 cagccaatgt cacggccgag ttcaaggaaa cgaggaggcg gctgagcgtg gagatctacg 2041 acaagttcca gcgtgccaca tccgtgaagc ggaagctctc cgcagagctg gcgggcaacc 2101 acaaccagga actgactccg tgtaggagga ccctgtctgt gaaccacctg accagcgaga 2161 gggaagtcct gcctcccttg ctgaaggctg agagcatcta tctgaacggt ctgacaccac 2221 actgtgctgg tgaggacata gctgtcattg agaacatgaa gacgcgtacg cggccgctcg 2281 agcagaaact catctcagaa gaggatctgg cagcaaatga tatcctggat tacaaggatg 2341 acgacgataa ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg 2401 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg 2461 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc 2521 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc 2581 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc 2641 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg 2701 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt 2761 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg 2821 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt 2881 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc 2941 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact 3001 cagtagatgc ctgttgaatt gggtacgcgg ccagcggcga gcggtatcag ctcactcaaa 3061 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 3121 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 3181 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 3241 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 3301 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 3361 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 3421 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 3481 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 3541 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 3601 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 3661 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 3721 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 3781 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 3841 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 3901 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc 3961 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg 4021 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg 4081 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc 4141 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc 4201 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac 4261 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag 4321 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca 4381 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg 4441 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg 4501 tcctgatagc gatccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca 4561 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg 4621 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct 4681 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg 4741 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc 4801 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc 4861 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc 4921 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc 4981 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct 5041 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg 5101 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg 5161 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa 5221 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat 5281 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg 5341 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat 5401 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg 5461 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca 5521 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc 5581 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 5641 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc 5701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 5761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 5821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 5881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 5941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 6001 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 6061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa tttt //