LOCUS       V030002                 6465 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030002
VERSION     V030002
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..2612
                     /label="Kcna3(NM_008418)"
                     /note="Kcna3(NM_008418)"
                     /gene="Kcna3"
     CDS             2631..2660
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2679..2702
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2722..2738)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2749..3371
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3520..4108)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4437..4484
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4716..5510)
                     /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(5545..5902)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5904..6008)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6035..6463
                     /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 gaccgtggtg cccggggacc acctgctgga gccagaggcg gcgggaggcg
     1081 gtggcgggga cccgcctcag ggaggctgtg gcagtggcgg cggcggtggc ggctgcgacc
     1141 gctacgagcc actgccaccc gcgctgcccg ccgcgggcga gcaagattgc tgcggcgagc
     1201 gtgtggtcat caacatctct gggctgcgct tcgagacgca gctcaagacc ctctgccagt
     1261 tccccgagac actgctgggc gaccccaagc ggcgcatgcg gtactttgac ccactccgca
     1321 atgagtactt cttcgaccgc aaccgaccca gcttcgacgc catcctctac tactaccagt
     1381 ccgggggccg cattcgccgg ccggtcaacg tgcccatcga catcttctcc gaggagatcc
     1441 gcttttacca gctgggtgag gaggccatgg aaaagttccg tgaggatgag ggcttcctgc
     1501 gggaggagga gcgacccctg ccccgccgtg acttccagcg ccaggtgtgg ctgctcttcg
     1561 aatatccgga gagctccggg ccggcccggg gcattgccat tgtgtcagtg ctggtcattc
     1621 tcatctccat tgtcatcttc tgcttggaga cgcttcccga gtttcgcgat gagaaagact
     1681 atcccgcctc cccgtcgcag gacgtgtttg aggctgccaa caacagcacg tcgggggccc
     1741 cttctggagc ctccagcttc tcggacccct tcttcgtggt ggagaccttg tgcatcatct
     1801 ggttctcctt tgagcttctg gtgcggttct ttgcttgccc cagtaaagcc accttctcca
     1861 gaaatatcat gaacttgata gacattgtgg ccatcattcc ttattttatc actctgggca
     1921 ctgagctggc tgaacgacaa ggtaatgggc agcaggccat gtcgctggcc atcctgagag
     1981 tcatccgcct agtaagggtt ttccgcatct tcaagctctc ccgccattct aaggggctgc
     2041 agatcctagg acagacgctg aaggcttcca tgcgggagct ggggctgctc atattcttcc
     2101 tcttcattgg ggtcatcctt ttctccagtg cagtctactt tgctgaggca gacgaccctt
     2161 cttcgggttt taacagtatc ccggatgcct tctggtgggc agtagtaacc atgacaactg
     2221 ttggttatgg tgatatgcac ccagtgacca taggaggcaa gattgtgggc tctctttgtg
     2281 ccatcgcagg tgtcttgacc attgcattgc cagttcctgt gattgtttcc aacttcaact
     2341 acttctacca ccgggagaca gaaggggaag agcaagccca gtacatgcac gtgggcagtt
     2401 gccagcacct ctcctcttca gccgaggagc tccgaaaagc ccggagtaac tccactctga
     2461 gtaagtcgga gtatatggtg atcgaagagg ggggtatgaa ccacagcgcc ttcccgcaga
     2521 cccccttcaa aacgggcaac tccacagcca cttgcaccac gaacaataac cccaactcct
     2581 gtgtcaacat caagaagata ttcactgatg tcacgcgtac gcggccgctc gagcagaaac
     2641 tcatctcaga agaggatctg gcagcaaatg atatcctgga ttacaaggat gacgacgata
     2701 aggtttaaac ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc
     2761 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2821 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     2881 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     2941 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     3001 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     3061 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3121 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3181 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     3241 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     3301 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     3361 cctgttgaat tgggtacgcg gccagcggcg agcggtatca gctcactcaa aggcggtaat
     3421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4261 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     4321 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     4381 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     4441 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     4501 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     4561 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     4621 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     4681 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     4741 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     4801 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     4861 cgatccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     4921 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     4981 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     5041 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     5101 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     5161 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     5221 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     5281 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     5341 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     5401 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     5461 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     5521 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     5581 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     5641 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     5701 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     5761 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     5821 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     5881 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
     5941 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6001 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     6061 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     6121 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6181 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6241 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6301 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     6361 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     6421 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga atttt
//
