LOCUS V030928 6156 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030928 VERSION V030928 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..2303 /label="Kcnj6(NM_010606)" /note="Kcnj6(NM_010606)" /gene="Kcnj6" CDS 2322..2351 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2370..2393 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(2413..2429) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 2440..3062 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3211..3799) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 4128..4175 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(4407..5201) /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(5236..5593) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(5595..5699) /gene="bla" /label="AmpR promoter" rep_origin 5726..6154 /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 gacaatggcc aagttaactg aatccatgac taacgtcttg gaaggcgatt 1081 ccatggacca ggatgtggaa agcccagtgg ccattcacca gccaaagttg cctaagcagg 1141 ccagggacga cctgccgaga cacatcagcc gagacaggac caaaaggaaa atccagaggt 1201 acgtgaggaa ggatgggaag tgcaacgttc accacggcaa tgtgcgggag acgtaccgat 1261 acctgacgga catcttcacc accctggtgg acctgaagtg gagattcaac ctgttgatct 1321 ttgtcatggt ctacacagtg acgtggcttt tctttgggat gatctggtgg ctgattgcgt 1381 acatccgggg agatatggac cacatagagg acccctcgtg gactccttgt gtcaccaacc 1441 tcaacgggtt tgtctctgct tttttattct ccatagagac agaaaccacc atcggttatg 1501 gctaccgggt catcacggac aagtgccctg aggggattat tctcctctta atccagtccg 1561 tgttggggtc cattgtcaac gccttcatgg taggatgtat gtttgtgaaa atatcccaac 1621 ccaagaagag ggcagagacc ctggtctttt ccacccacgc ggtgatctcc atgcgggatg 1681 ggaaactgtg cttgatgttc cgggtggggg acttgaggaa ttctcacatt gtggaggcat 1741 ccatcagagc caagttgatc aagtccaaac agacttcaga gggggagttt attcccctca 1801 accagactga tatcaacgtg gggtactaca caggggacga ccggctcttt ctggtgtcac 1861 cattgattat tagccatgaa attaaccaac agagtccctt ctgggagatc tccaaagcgc 1921 agctgcctaa agaggaactg gagattgtgg tcatcctgga gggaatggtg gaagccacag 1981 gaatgacgtg ccaagcccga agctcctaca tcaccagtga gatcttgtgg ggttaccggt 2041 tcacacctgt cctaacgctg gaagacgggt tctacgaagt tgactacaac agcttccatg 2101 agacctatga gaccagcacc ccgtccctta gtgccaaaga gctagcggag ctggctaacc 2161 gggcagagct gcctctgagt tggtctgtgt ccagcaaact gaaccaacat gcagaattgg 2221 agacagaaga ggaagagaag aacccggaag aactgacgga gaggaatggt gacgtggcaa 2281 acctagagaa tgaatccaaa gtaacgcgta cgcggccgct cgagcagaaa ctcatctcag 2341 aagaggatct ggcagcaaat gatatcctgg attacaagga tgacgacgat aaggtttaaa 2401 cggccggccg cggtcatagc tgtttcctga acagatcccg ggtggcatcc ctgtgacccc 2461 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta 2521 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg 2581 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc 2641 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc 2701 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg 2761 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg 2821 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga 2881 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag 2941 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg 3001 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa 3061 ttgggtacgc ggccagcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 3121 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 3181 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 3241 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 3301 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 3361 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 3421 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 3481 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 3541 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 3601 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 3661 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 3721 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 3781 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 3841 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 3901 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga 3961 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact 4021 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg 4081 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc 4141 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat 4201 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag 4261 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct 4321 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc 4381 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg 4441 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg 4501 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcgatccgcc 4561 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc 4621 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg 4681 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga 4741 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg 4801 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg 4861 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc 4921 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg 4981 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg 5041 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg 5101 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa 5161 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct 5221 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg 5281 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc 5341 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg 5401 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga 5461 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct 5521 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt 5581 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt 5641 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 5701 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt 5761 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 5821 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 5881 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 5941 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 6001 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 6061 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 6121 aatgagctga tttaacaaaa atttaacgcg aatttt //