LOCUS       V031354                 5472 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031354
VERSION     V031354
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..1619
                     /label="Cdkn1b(NM_009875)"
                     /note="Cdkn1b(NM_009875)"
                     /gene="Cdkn1b"
     CDS             1638..1667
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             1686..1709
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(1729..1745)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    1756..2378
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(2527..3115)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3444..3491
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(3723..4517)
                     /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(4552..4909)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(4911..5015)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5042..5470
                     /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 gtcaaacgtg agagtgtcta acgggagccc gagcctggag cggatggacg
     1081 ccagacaagc ggatcacccc aagccttccg cctgcagaaa tctcttcggc ccggtcaatc
     1141 atgaagaact aacccgggac ttggagaagc actgccggga tatggaagaa gcgagtcagc
     1201 gcaagtggaa tttcgacttt cagaatcata agcccctgga gggcagatac gaatggcagg
     1261 aggtggagag gggcagcttg cccgagttct actacaggcc cccgcgcccc cccaagagcg
     1321 cctgcaaggt gctggcgcag gagagccagg atgtcagcgg gagccgccag gcggtgcctt
     1381 taattgggtc tcaggcaaac tctgaggacc ggcatttggt ggaccaaatg cctgactcgt
     1441 cagacaatca ggctgggtta gcggagcagt gtccagggat gaggaagcga cctgctgcag
     1501 aagattcttc ttcgcaaaac aaaagggcca acagaacaga agaaaatgtt tcagacggtt
     1561 ccccgaacgc tggcactgtg gagcagacgc ccaagaagcc cggccttcga cgccagacga
     1621 cgcgtacgcg gccgctcgag cagaaactca tctcagaaga ggatctggca gcaaatgata
     1681 tcctggatta caaggatgac gacgataagg tttaaacggc cggccgcggt catagctgtt
     1741 tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     1801 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     1861 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     1921 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     1981 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     2041 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     2101 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     2161 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     2221 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     2281 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     2341 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcggcgagc
     2401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2941 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3241 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc
     3301 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga
     3361 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc
     3421 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct
     3481 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     3541 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     3601 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     3661 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     3721 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     3781 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     3841 cgggtagcca acgctatgtc ctgatagcga tccgccacac ccagccggcc acagtcgatg
     3901 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     3961 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc
     4021 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     4081 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     4141 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     4201 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     4261 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     4321 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     4381 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     4441 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     4501 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     4561 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     4621 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     4681 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     4741 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     4801 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4861 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     4921 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4981 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5041 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5101 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5161 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5221 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5281 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5341 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5401 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5461 aacgcgaatt tt
//
