LOCUS       V031327                 6972 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031327
VERSION     V031327
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..3119
                     /label="Trf(NM_133977)"
                     /note="Trf(NM_133977)"
                     /gene="Trf"
     CDS             3138..3167
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3186..3209
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3229..3245)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3256..3878
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4027..4615)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4944..4991
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5223..6017)
                     /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(6052..6409)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6411..6515)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6542..6970
                     /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 gaggctcacc gtgggtgccc tgctggcctg cgctgccctg gggctgtgtc
     1081 tggctgtccc tgacaaaacg gtcaaatggt gcgcagtgtc agagcacgag aataccaaat
     1141 gcatcagctt ccgtgaccac atgaagaccg tccttccgcc tgatggcccc cggcttgcct
     1201 gtgtgaagaa aacctcctat ccggattgca tcaaggccat ttctgcaagt gaagccgatg
     1261 ctatgacctt ggatgggggt tgggtgtacg atgccggcct gactccgaac aacctgaagc
     1321 ccgtggcggc ggagttttat ggatcagtgg aacatccaca gacctactac tacgctgtgg
     1381 ctgtggtaaa gaagggaaca gacttccagc tgaaccagct cgaaggcaag aagtcctgcc
     1441 acacaggcct gggaaggtct gcaggctggg tcatccccat tggcttgctc ttctgtaagc
     1501 tgtcggagcc ccgcagtcct cttgagaaag ctgtgtccag tttcttctcg ggcagttgtg
     1561 tcccctgtgc agatccagtg gccttcccca aactgtgtca actgtgccca ggctgtggct
     1621 gctcctccac tcaaccgttc tttggctacg taggcgcatt caagtgtctg aaagatggcg
     1681 gtggggatgt ggcctttgtc aagcacacaa ccatatttga ggtcttgccg gagaaggctg
     1741 acagggacca atatgaactg ctctgccttg acaatacccg caagccagtg gatcagtatg
     1801 aggattgcta cctggctcgg atcccttctc atgctgttgt ggctcgaaaa aacaatggca
     1861 aggaagactt gatctgggag attctcaaag tggcacagga acactttggc aaaggcaaat
     1921 caaaagactt ccaactgttc agctctcctc ttgggaaaga cctgctgttt aaagattctg
     1981 cctttgggct gttaagggtc cccccaagga tggactacag gctgtacctt ggccataact
     2041 atgtcactgc cattcggaat cagcaggaag gcgtgtgccc ggagggctcg atcgacaact
     2101 cgccagtgaa gtggtgtgca ctgagtcacc tggagagaac caagtgtgac gagtggagca
     2161 tcatcagtga gggaaagata gagtgtgagt cagcagagac cactgaggac tgcattgaaa
     2221 agattgtgaa cggagaagcg gacgccatga ctttggatgg aggacatgcc tacattgcag
     2281 gccagtgtgg tctagtgcct gtcatggcag agtactacga gagctctaat tgtgccatcc
     2341 catcacaaca aggtatcttt cctaaagggt attatgccgt ggctgtggtg aaggcatcgg
     2401 acactagcat cacctggaac aacctgaaag gcaagaagtc ctgccacact ggggtagaca
     2461 gaaccgctgg ttggaacatc cctatgggca tgctgtacaa caggatcaac cactgcaaat
     2521 tcgatgaatt tttcagtcaa ggctgcgctc ccgggtatga gaagaattcc accctctgtg
     2581 acctgtgtat tggcccactc aaatgtgctc cgaacaacaa agaggaatat aatggttaca
     2641 caggggcttt caggtgtctc gttgagaaag gagatgtagc ctttgtgaaa caccagactg
     2701 tcctggataa caccgaagga aagaaccctg ccgaatgggc taagaatctg aagcaggaag
     2761 acttcgagtt gctctgccct gatggcacca ggaagcctgt gaaagatttt gccagctgcc
     2821 acctggccca agctccaaac catgttgtgg tctcacgaaa agagaaggca gcccgggtta
     2881 aggctgtact gactagccag gagactttat ttgggggaag tgactgcacc ggcaatttct
     2941 gtttgttcaa gtctaccacc aaggaccttc tgttcaggga tgacaccaaa tgtttcgtta
     3001 aacttccaga gggtaccaca cctgaaaaat acttaggagc ggagtacatg caatctgtcg
     3061 gtaacatgag gaagtgctca acctcacgac tcctggaagc ctgcactttc cacaaacata
     3121 cgcgtacgcg gccgctcgag cagaaactca tctcagaaga ggatctggca gcaaatgata
     3181 tcctggatta caaggatgac gacgataagg tttaaacggc cggccgcggt catagctgtt
     3241 tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     3301 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     3361 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     3421 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     3481 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     3541 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     3601 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     3661 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     3721 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     3781 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     3841 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcggcgagc
     3901 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3961 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4441 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4741 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc
     4801 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga
     4861 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc
     4921 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct
     4981 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     5041 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     5101 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     5161 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     5221 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     5281 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     5341 cgggtagcca acgctatgtc ctgatagcga tccgccacac ccagccggcc acagtcgatg
     5401 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     5461 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc
     5521 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     5581 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     5641 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     5701 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     5761 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     5821 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     5881 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     5941 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     6001 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     6061 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     6121 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6181 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6241 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     6301 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     6361 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     6421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6481 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6541 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6601 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6661 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6721 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6781 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6841 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6901 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6961 aacgcgaatt tt
//
