LOCUS       V031202                 6276 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031202
VERSION     V031202
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..2423
                     /label="Gck(NM_010292)"
                     /note="Gck(NM_010292)"
                     /gene="Gck"
     CDS             2442..2471
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2490..2513
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2533..2549)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2560..3182
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3331..3919)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4248..4295
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4527..5321)
                     /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(5356..5713)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5715..5819)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5846..6274
                     /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 gctggatgac agagccagga tggaggccac caagaaggaa aaggtagagc
     1081 agatcctggc agagttccag ctgcaggagg aagacctgaa gaaggtgatg agccggatgc
     1141 agaaggagat ggaccgtggc ctgaagctgg agacccatca ggaggccagt gtaaagatgt
     1201 tgcccaccta cgtgcgttcc accccagaag gctcagaagt tggagacttt ctctccttag
     1261 acctgggagg aaccaacttc agggtgatgc tggtgaaagt gggggagggg gaggcaggac
     1321 agtggagcgt gaagacgaaa caccagatgt attccatccc cgaggacgcc atgacgggca
     1381 ctgcggagat gctctttgac tacatctctg agtgcatctc tgacttcctg gacaagcatc
     1441 agatgaaaca caagaaacta cccctgggct tcaccttctc cttccctgta aggcacgaag
     1501 acatagacaa gggcatcctg ctcaactgga ccaagggctt caaggcctcc ggagcagaag
     1561 ggaacaacat cgtgggactt ctccgagatg ctatcaagag gagaggggac tttgagatgg
     1621 atgtggtggc aatggtgaat gacacggtgg ccacaatgat ctcctgctac tatgaagacc
     1681 gccaatgtga ggtcggcatg attgtgggca ccggctgcaa cgcctgctac atggaggaga
     1741 tgcagaatgt ggagctggtg gaaggcgatg aggggcgcat gtgtgtcaac acagagtggg
     1801 gcgccttcgg gaactccggt gagctggacg agttcctcct ggagtacgac cggatggtgg
     1861 atgagagctc agtgaacccc ggtcagcagc tgtacgaaaa gatcattggc ggaaagtaca
     1921 tgggcgagct ggtacgactt gtgctgctca agctggtaga ggagaatctt ctgttccacg
     1981 gagaggcctc agagcagctg cgcacacgtg gtgcttttga gacccgtttt gtgtcgcagg
     2041 tggagagcga ctctggggac cgaaggcaga tccttaacat cctgagcact ctgggccttc
     2101 gaccctctgt cgccgactgc gacattgtgc gccgtgcctg tgaaagcgtg tccactcgcg
     2161 ccgcccacat gtgctcagca ggactagcgg gggtcataaa tcgcatgcgc gaaagccgca
     2221 gtgaggacgt gatgcgcatc acggtgggcg tggatggctc cgtgtacaag ctgcacccga
     2281 gcttcaagga gcggtttcac gccagtgtgc gcaggctgac acccaactgc gaaatcacct
     2341 tcattgaatc agaggagggc agcggcaggg gagccgcact ggtctctgcg gtggcctgca
     2401 agaaggcttg catgctgggc cagacgcgta cgcggccgct cgagcagaaa ctcatctcag
     2461 aagaggatct ggcagcaaat gatatcctgg attacaagga tgacgacgat aaggtttaaa
     2521 cggccggccg cggtcatagc tgtttcctga acagatcccg ggtggcatcc ctgtgacccc
     2581 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     2641 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     2701 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     2761 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     2821 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     2881 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     2941 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3001 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag
     3061 caggaggacg tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg
     3121 agttgcttgc ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa
     3181 ttgggtacgc ggccagcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     4081 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     4141 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     4201 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     4261 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     4321 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     4381 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     4441 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     4501 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     4561 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     4621 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcgatccgcc
     4681 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     4741 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     4801 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     4861 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     4921 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     4981 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     5041 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     5101 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     5161 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     5221 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     5281 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     5341 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     5401 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     5461 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     5521 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5581 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     5641 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     5701 ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5761 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5821 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5881 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5941 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6001 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6061 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6121 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6181 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6241 aatgagctga tttaacaaaa atttaacgcg aatttt
//
