LOCUS       V031204                 8511 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031204
VERSION     V031204
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..4658
                     /label="Egfr(NM_207655)"
                     /note="Egfr(NM_207655)"
                     /gene="Egfr"
     CDS             4677..4706
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4725..4748
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4768..4784)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4795..5417
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5566..6154)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6483..6530
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6762..7556)
                     /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(7591..7948)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7950..8054)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8081..8509
                     /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 gcgaccctca gggaccgcga gaaccacact gctggtgttg ctgaccgcgc
     1081 tctgcgccgc aggtggggcg ttggaggaaa agaaagtctg ccaaggcaca agtaacaggc
     1141 tcacccaact gggcactttt gaagaccact ttctgagcct gcagaggatg tacaacaact
     1201 gtgaagtggt ccttgggaac ttggaaatta cctatgtgca aaggaattac gacctttcct
     1261 tcttaaagac catccaggag gtggccggct atgtcctcat tgccctcaac accgtggaga
     1321 gaatcccttt ggagaacctg cagatcatca ggggaaatgc tctttatgaa aacacctatg
     1381 ccttagccat cctgtccaac tatgggacaa acagaactgg gcttagggaa ctgcccatgc
     1441 ggaacttaca ggaaatcctg attggtgctg tgcgattcag caacaacccc atcctctgca
     1501 atatggatac tatccagtgg agggacatcg tccaaaacgt ctttatgagc aacatgtcaa
     1561 tggacttaca gagccatccg agcagttgcc ccaaatgtga tccaagctgt cccaatggaa
     1621 gctgctgggg aggaggagag gagaactgcc agaaattgac caaaatcatc tgtgcccagc
     1681 aatgttccca tcgctgtcgt ggcaggtccc ccagtgactg ctgccacaac caatgtgctg
     1741 cggggtgtac agggccccga gagagtgact gtctggtctg ccaaaagttc caagatgagg
     1801 ccacatgcaa agacacctgc ccaccactca tgctgtacaa ccccaccacc tatcagatgg
     1861 atgtcaaccc tgaagggaag tacagctttg gtgccacctg tgtgaagaag tgcccccgaa
     1921 actacgtggt gacagatcat ggctcatgtg tccgagcctg tgggcctgac tactacgaag
     1981 tggaagaaga tggcatccgc aagtgtaaaa aatgtgatgg gccctgtcgc aaagtttgta
     2041 atggcatagg cattggtgaa tttaaagaca cactctccat aaatgctaca aacatcaaac
     2101 acttcaaata ctgcactgcc atcagcgggg accttcacat cctgccagtg gcctttaagg
     2161 gggattcttt cacgcgcact cctcctctag acccacgaga actagaaatt ctaaaaaccg
     2221 taaaggaaat aacaggcttt ttgctgattc aggcttggcc tgataactgg actgacctcc
     2281 atgctttcga gaacctagaa ataatacgtg gcagaacaaa gcaacatggt cagttttctt
     2341 tggcggtcgt tggcctgaac atcacatcac tggggctgcg ttccctcaag gagatcagtg
     2401 atggggatgt gatcatttct ggaaaccgaa atttgtgcta cgcaaacaca ataaactgga
     2461 aaaaactctt cgggacaccc aatcagaaaa ccaaaatcat gaacaacaga gctgagaaag
     2521 actgcaaggc cgtgaaccac gtctgcaatc ctttatgctc ctcggaaggc tgctggggcc
     2581 ctgagcccag ggactgtgtc tcctgccaga atgtgagcag aggcagggag tgcgtggaga
     2641 aatgcaacat cctggagggg gaaccaaggg agtttgtgga aaattctgaa tgcatccagt
     2701 gccatccaga atgtctgccc caggccatga acatcacctg tacaggcagg ggaccagaca
     2761 actgcatcca gtgtgcccac tacattgatg gcccacactg tgtcaagacc tgcccagctg
     2821 gcatcatggg agagaacaac actctggtct ggaagtatgc agatgccaat aatgtctgcc
     2881 acctatgcca cgccaactgt acctatggat gtgctgggcc aggtcttcaa ggatgtgaag
     2941 tgtggccatc tgggccaaag ataccatcta ttgccactgg gattgtgggt ggcctcctct
     3001 tcatagtggt ggtggccctt gggattggcc tattcatgcg aagacgtcac attgttcgaa
     3061 agcgtacact acgccgcctg cttcaagaga gagagctcgt ggaacctctc acacccagcg
     3121 gagaagctcc aaaccaagcc cacttgagga tattaaagga aacagaattc aaaaagatca
     3181 aagttctggg ttcgggagca tttggcacag tgtataaggg tctctggatc ccagaaggtg
     3241 agaaagtaaa aatcccggtg gccatcaagg agttaagaga agccacatct ccaaaagcca
     3301 acaaagaaat ccttgacgaa gcctatgtga tggctagtgt ggacaaccct catgtatgcc
     3361 gcctcctggg catctgtctg acctccactg tccagctcat tacacagctc atgccctacg
     3421 gttgcctcct ggactacgtc cgagaacaca aggacaacat tggctcccag tacctcctca
     3481 actggtgtgt gcagattgca aagggcatga actacctgga agatcggcgt ttggtgcacc
     3541 gtgacttggc agccaggaat gtactggtga agacaccaca gcatgtcaag atcacagatt
     3601 ttgggctggc caaactgctt ggtgctgaag agaaagaata tcatgccgag gggggcaaag
     3661 tgcctatcaa gtggatggct ttggaatcaa ttttacaccg aatttataca caccaaagtg
     3721 atgtctggag ctatggtgtc actgtgtggg aactgatgac ctttgggtcc aagccttatg
     3781 atggaatccc agcaagtgac atctcatcca tcctagagaa aggagagcgc cttccacagc
     3841 cacctatctg caccatcgat gtctacatga tcatggtcaa gtgctggatg atagatgctg
     3901 atagccgccc aaagttccga gagttgattc ttgaattctc caaaatggcc cgagacccac
     3961 agcgctacct tgttatccag ggggatgaaa gaatgcattt gccaagccct acagactcca
     4021 acttttaccg agccctgatg gatgaagagg acatggagga tgtagttgat gctgatgagt
     4081 atcttatccc acagcaaggc ttcttcaaca gcccgtccac gtcgaggact cccctcttga
     4141 gttctctgag tgcaactagc aacaattcca ctgtggcttg cattaataga aatgggagct
     4201 gccgtgtcaa agaagacgcc ttcttgcagc ggtacagctc cgaccccaca ggtgctgtaa
     4261 cagaggacaa catagatgac gcattcctcc ctgtacctga atatgtaaac caatctgttc
     4321 ccaagaggcc agcaggctct gtgcagaacc ctgtctatca caatcagccc ctgcatccag
     4381 ctcctggaag agacctgcat tatcaaaatc cccacagcaa tgcagtgggc aaccctgagt
     4441 atctcaacac tgcccagcct acctgtctca gtagtgggtt taacagccct gcactctgga
     4501 tccagaaagg cagtcaccaa atgagcctag acaaccctga ctaccagcag gacttcttcc
     4561 ccaaggaaac caagccaaat ggcatattta agggccccac agctgaaaat gcagagtacc
     4621 tacgggtggc acctccaagc agtgagttta ttggagcaac gcgtacgcgg ccgctcgagc
     4681 agaaactcat ctcagaagag gatctggcag caaatgatat cctggattac aaggatgacg
     4741 acgataaggt ttaaacggcc ggccgcggtc atagctgttt cctgaacaga tcccgggtgg
     4801 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4861 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4921 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4981 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     5041 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     5101 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     5161 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     5221 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta
     5281 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa
     5341 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag
     5401 tagatgcctg ttgaattggg tacgcggcca gcggcgagcg gtatcagctc actcaaaggc
     5461 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5521 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5581 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5641 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5701 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5761 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5821 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5881 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5941 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6001 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6061 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6121 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6181 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6301 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
     6361 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
     6421 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     6481 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     6541 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa
     6601 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     6661 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     6721 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     6781 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     6841 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     6901 tgatagcgat ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     6961 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     7021 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     7081 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     7141 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     7201 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     7261 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     7321 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
     7381 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     7441 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     7501 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     7561 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc
     7621 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7681 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7741 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     7801 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     7861 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     7921 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt
     7981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     8101 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     8161 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     8221 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     8281 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     8341 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     8401 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     8461 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt t
//
