LOCUS       V029736                 6852 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029736
VERSION     V029736
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..2999
                     /label="Ppp1r15a(NM_008654)"
                     /note="Ppp1r15a(NM_008654)"
                     /gene="Ppp1r15a"
     CDS             3018..3047
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3066..3089
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3109..3125)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3136..3758
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3907..4495)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4824..4871
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5103..5897)
                     /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(5932..6289)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6291..6395)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6422..6850
                     /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 ggccccgagc ccaagacccc agcatgtcct gcactggagg gacgcccaca
     1081 acttctatct cctgtcccca ctgatgggct tgctcagtcg ggcctggagc cgcctgaggg
     1141 gcccagaagt cccagaggca tggctggcaa aaacagtaac aggagcagat cagatagaag
     1201 ctgcggctct gctgacacct acccctgtct ctggtaacct cctccctcat ggggagactg
     1261 aagaaagtgg atctcctgaa cagagtcaag cagcccagag gctctgcctt gtggaagctg
     1321 aaagttcccc tcctgaaact tggggacttt caaatgttga tgagtacaat gcaaagccag
     1381 gacaagatga ccttagagag aaggaaatgg aacgcacagc tggcaaggcc acactacagc
     1441 ccgctggcct gcaaggggct gataagaggc ttggggaggt ggtggctaga gaagagggag
     1501 tggctgagcc cgcttatccc acatcacagc tggagggtgg tccagctgag aatgaagagg
     1561 atggagaaac agtgaagact taccaagctt ctgctgcttc catagctccg ggatacaaac
     1621 ccagcacccc tgtgcctttc ttgggggagg cagaacatca agccacggaa gaaaaaggaa
     1681 cagaaaacaa ggctgacccc tccaactctc cttcttcagg ctcccactcc agagcctggg
     1741 agtactactc tagagagaag cctaagcagg agggagaagc caaggtagag gcacacaggg
     1801 cagggcaggg tcacccttgt cggaatgctg aggctgagga aggaggacct gagacaactt
     1861 ttgtctgtac tggaaatgcc ttcctgaagg cctgggtgta tcgcccagga gaggacacag
     1921 aggaagaaga caacagcgat tcggattcag ctgaggaaga cacagctcag accggtgcca
     1981 ccccccatac aagtgccttc ctgaaggcct gggtgtatcg cccaggagag gacacagagg
     2041 aagaagacag cgattcggat tcagctgagg aagacacagc tcagaccggt gccacccccc
     2101 atacaagtgc cttcctgaag gcctgggtgt atcgcccagg agaggacaca gaggaagaaa
     2161 acagcgattt ggattcagct gaggaagaca cagctcagac cggtgccacc ccccatacaa
     2221 gtgccttcct gaaggcctgg gtgtatcgcc caggagagga cacagaggaa gaaaacagcg
     2281 atttggattc agctgaggaa gacacagctc agaccggtgc caccccacat acaagtccct
     2341 tcctgaaggc ctgggtgtat cgcccaggag aggacacaga agatgacaca gaagaggaag
     2401 aggacagtga gaatgtggcc ccaggtgact cagaaacagc tgactcaagc cagagtccct
     2461 gccttcagcc ccagcgttgt ctaccaggag agaagaccaa gggacgtggg gaagagcccc
     2521 ctctcttcca ggtggccttc tatttacccg gagagaagcc agaatcacct tgggctgcac
     2581 ctaagctgcc ccttcgactg cagaggcggc tcagattgtt caaagccccc acccgggatc
     2641 aggaccccga gattcctcta aaagctcgga aggtacactt cgctgagaaa gtcacagtcc
     2701 atttccttgc tgtctgggca ggaccagccc aagctgcccg tcgaggtccc tgggagcagt
     2761 ttgcacgaga tcgaagccgc tttgctcgac gcattgccca ggcagaggag aagctgggtc
     2821 cctaccttac ccctgattcc agggccagag catgggcacg ccttagaaac ccatctcttc
     2881 cacagtccga gcctcgctct tcctctgagg ccactccctt gacccaagat gtgaccacac
     2941 cctctcccct tcccagtgaa accccttcgc ccagcctgta cttgggaggg aggcggggca
     3001 cgcgtacgcg gccgctcgag cagaaactca tctcagaaga ggatctggca gcaaatgata
     3061 tcctggatta caaggatgac gacgataagg tttaaacggc cggccgcggt catagctgtt
     3121 tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     3181 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     3241 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     3301 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     3361 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     3421 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     3481 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     3541 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     3601 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     3661 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     3721 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcggcgagc
     3781 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3841 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4321 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4621 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc
     4681 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga
     4741 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc
     4801 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct
     4861 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     4921 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     4981 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     5041 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     5101 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     5161 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     5221 cgggtagcca acgctatgtc ctgatagcga tccgccacac ccagccggcc acagtcgatg
     5281 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     5341 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc
     5401 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     5461 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     5521 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     5581 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     5641 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     5701 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     5761 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     5821 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     5881 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     5941 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     6001 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6061 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6121 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     6181 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     6241 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     6301 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6361 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6421 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6481 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6541 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6601 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6661 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6721 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6781 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6841 aacgcgaatt tt
//
