LOCUS       V028725                 6846 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028725
VERSION     V028725
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..2993
                     /label="Cacnb2(NM_023116)"
                     /note="Cacnb2(NM_023116)"
                     /gene="Cacnb2"
     CDS             3012..3041
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3060..3083
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3103..3119)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3130..3752
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3901..4489)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4818..4865
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5097..5891)
                     /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(5926..6283)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6285..6389)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6416..6844
                     /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 ggtccaaagc gacacgtcca agtcgcctcc cgtagcggcg gtggcgcagg
     1081 agagccagat ggagctgctg gagagcgcgg ctcccgcggg ggcgctcgga gcacagtcct
     1141 atggaaaagg agccaggagg aaaaatagat ttaaaggctc ggatggaagc acatcgtcag
     1201 acactacctc aaacagtttt gtccgccagg gttcggcaga ctcctacacc agccgcccat
     1261 ccgattcaga tgtgtctttg gaagaggacc gggaggcagt acgtagagaa gctgagcggc
     1321 aggcacaggc acagttggaa aaagcaaaga caaaacctgt tgcatttgca gttcggacca
     1381 atgtcagata cagcgcagcc caggaggatg acgttccggt gcctggcatg gccatctcct
     1441 tcgaggccaa agattttctg catgttaaag aaaaatttaa taatgactgg tggataggac
     1501 ggctggttaa agaaggctgt gaaatcggat ttattccaag cccagtcaaa ctagagaaca
     1561 tgaggctaca gcacgaacag agagccaagc aagggaaatt ctattccagt aaatcaggag
     1621 ggaattcatc atccagtttg ggtgacatag tgcccagttc cagaaaatcg acaccgccgt
     1681 catctgctat agacatagat gctactggct tagatgcaga agagaatgat attccagcaa
     1741 accaccgctc ccctaagccc agtgcaaaca gtgtaacatc accccactcc aaagagaaaa
     1801 gaatgccctt ctttaagaag acagagcaca ctcctcctta tgatgtggta ccttccatgc
     1861 gaccagtggt gctggtgggc ccgtctctga aggggtatga ggtcacagat atgatgcaaa
     1921 aagcactgtt tgattttcta aaacacagat ttgaagggcg gatatccatc acaagagtca
     1981 ctgctgacat ctcccttgcc aaacgctcag tattaaacaa tcccagtaag catgcaatca
     2041 tagaaagatc caacacaagg tccagcttag cggaagttca gagtgaaatt gaaaggattt
     2101 ttgaacttgc aagaacactg caattggtag tccttgacgc ggatacaatt aatcacccag
     2161 ctcaactcag taagacctct ttggccccta tcatagtgta cgtaaagatt tcttctccca
     2221 aggttttaca aaggttgata aaatctcgag ggaaatctca agcaaaacac ctcaatgtcc
     2281 agatggtagc agctgataaa ctggcccaat gtcctccgca ggaatcattt gatgtgatct
     2341 tggatgagaa ccagctggag gacgcttgtg agcatcttgc cgactatctg gaggcatact
     2401 ggaaggccac ccaccctccc agcggtaacc tccccaaccc tctgcttagc cggactttag
     2461 cctcctcaac tttacctctg agccccaccc tcgcctctaa ttcacagggt tctcaaggtg
     2521 atcaaaggcc tgatcgctct gctccccggt ctgcttccca agctgaagaa gaaccttgcc
     2581 tagaacccgt caaaaaatcc caacaccgtt cctcctcagc cacacaccaa aaccaccgca
     2641 gcgggacagg tcgaggcctc tctaggcaag agacgtttga ctctgaaacc caagagagcc
     2701 gagactctgc ctacgtggag ccaaaggaag attattcaca tgaacatgtg gaccggtatg
     2761 tcccacaccg cgagcataac cacagagagg agacccacag cagcaatggc cacagacaca
     2821 gggagtctcg ccaccgctct agggacatgg gtcgagacca ggaccacaat gagtgcatca
     2881 aacaacgaag ccggcataaa tctaaggatc gctactgtga caaggagggg gaagtaatat
     2941 ccaagagaag gaatgaggct ggcgagtgga acagggatgt atacatccgc caaacgcgta
     3001 cgcggccgct cgagcagaaa ctcatctcag aagaggatct ggcagcaaat gatatcctgg
     3061 attacaagga tgacgacgat aaggtttaaa cggccggccg cggtcatagc tgtttcctga
     3121 acagatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3181 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3241 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3301 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     3361 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     3421 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     3481 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3541 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3601 gtccttctga ttttaaaata actataccag caggaggacg tccagacaca gcataggcta
     3661 cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc
     3721 gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcggc gagcggtatc
     3781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4321 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4621 atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt
     4681 ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac
     4741 gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga
     4801 ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat
     4861 tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc
     4921 cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     4981 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     5041 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     5101 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     5161 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     5221 gccaacgcta tgtcctgata gcgatccgcc acacccagcc ggccacagtc gatgaatcca
     5281 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     5341 agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc
     5401 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     5461 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     5521 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     5581 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5641 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5701 gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5761 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5821 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5881 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag
     5941 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     6001 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6061 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     6121 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     6181 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6241 ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa
     6301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     6421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6541 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6781 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6841 aatttt
//
