LOCUS       V031133                 6441 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031133
VERSION     V031133
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..2588
                     /label="Was(NM_009515)"
                     /note="Was(NM_009515)"
                     /gene="Was"
     CDS             2607..2636
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2655..2678
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2698..2714)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2725..3347
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3496..4084)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4413..4460
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4692..5486)
                     /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(5521..5878)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5880..5984)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6011..6439
                     /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 gaatagtggc cctggccctg taggaggcag gcctgggggc cgagggggac
     1081 cagccgttca gcagaacatt ccttccaacc tcctccagga ccatgaaaac cagagactct
     1141 ttgagcttct tggccgaaaa tgctggacac tggctaccac agttgttcag ctctacctgg
     1201 cactgccccc tggagctgag cactggacca tggaacactg cggggctgtg tgcttcgtga
     1261 aggataaccc tcagaagtcc tacttcatcc gcctttatgg cctacaggct ggtcggctac
     1321 tctgggaaca ggagctgtac tctcagctgg tttatctcac tcccaccccg ttcttccaca
     1381 cttttgctgg agatgactgt caagtaggac tgaactttgc ggatgagagt gaagcccagg
     1441 ccttccgggc cttggtgcag gagaagatac aaaaaaggaa tcagaggcaa agcggagaaa
     1501 gacgccagct accaccacca ccagcaccaa tcaatgagga gagaagagga gggctcccac
     1561 ctgtgccccc acacccgggt ggagatcatg ggggcccatc aggtggtcca ctatctctag
     1621 gacttgtgac ggtcgacatt cagaaccctg acatcacaag ttcacgatac cgtgggctcc
     1681 ctgcacctgg ccctggccca actgataaga aacgctcagg gaaaaagaag atcagcaaag
     1741 ctgatatcgg agcaccgagt ggattcaaac atgtcagcca cgtgggctgg gatccccaga
     1801 atggatttga tgtgaacaac ctagacccgg atctgcggag cttgttctcc agggcaggaa
     1861 tcagcgaggc ccagctcact gacgcagaga cctccaagct catctacgat tttattgagg
     1921 accagggagg tctagaggct gtccggcagg agatgaggcg ccaagagcca ctcccaccac
     1981 ctccgccgcc atgcagagga ggaggaggag gaggaggagg aggaggagga ggaggaggag
     2041 gaggaggagg ccagcctctg agacctcctg ttgtggggag taataagggt cgctcaggtc
     2101 cactgccccc tgtacctatg gggggtgccc cacctccacc aacaccacga gggcccccac
     2161 caccaggccg agggggtcct cctccaccac cccctccagc cactggacga tctggaccac
     2221 cacctcctcc actccctgga gctgggggac caccagcacc gccaccacca ccaccaccac
     2281 caccacctcc accctgccct gggagtgggc ccgcccctcc cccgctccct cctactccag
     2341 tgtctggggg gagcccagca cctggtgggg gccggggtgc acttttggac caaatccggc
     2401 agggaattca gctgaacaag acccctggag ctctagagaa ctcagtacag caaccacccg
     2461 cgcagcagtc agaaggccta gtaggtgccc tgatgcatgt catgcagaag aggagtagag
     2521 tcatccattc ctcagatgaa ggggaggatc agaccggcga ggatgaagag gatgatgaat
     2581 gggatgacac gcgtacgcgg ccgctcgagc agaaactcat ctcagaagag gatctggcag
     2641 caaatgatat cctggattac aaggatgacg acgataaggt ttaaacggcc ggccgcggtc
     2701 atagctgttt cctgaacaga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     2761 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     2821 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     2881 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     2941 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3001 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3061 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3121 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3181 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     3241 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     3301 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     3361 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3661 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3901 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4081 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4141 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4201 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     4261 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     4321 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     4381 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     4441 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     4501 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     4561 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     4621 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     4681 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     4741 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     4801 gcaatatcac gggtagccaa cgctatgtcc tgatagcgat ccgccacacc cagccggcca
     4861 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     4921 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     4981 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     5041 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     5101 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     5161 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     5221 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     5281 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     5341 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     5401 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     5461 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     5521 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     5581 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     5641 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     5701 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     5761 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     5821 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     5881 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5941 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6001 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6061 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6121 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6181 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6241 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6301 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6361 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6421 caaaaattta acgcgaattt t
//
