LOCUS       V026824                 5442 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026824
VERSION     V026824
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..1589
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /transl_table=1
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
                     /label="DHFR"
     CDS             1608..1637
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             1656..1679
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(1699..1715)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    1726..2348
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(2497..3085)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3414..3461
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(3693..4487)
                     /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(4522..4879)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(4881..4985)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5012..5440
                     /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 ggttcgacca ttgaactgca tcgtcgccgt gtcccaaaat atggggattg
     1081 gcaagaacgg agacctaccc tggcctccgc tcaggaacga gttcaagtac ttccaaagaa
     1141 tgaccacaac ctcttcagtg gaaggtaaac agaatctggt gattatgggt aggaaaacct
     1201 ggttctccat tcctgagaag aatcgacctt taaaggacag aattaatata gttctcagta
     1261 gagaactcaa agaaccacca cgaggagctc attttcttgc caaaagtttg gatgatgcct
     1321 taagacttat tgaacaaccg gaattggcaa gtaaagtaga catggtttgg atagtcggag
     1381 gcagttctgt ttaccaggaa gccatgaatc aaccaggcca cctcagactc tttgtgacaa
     1441 ggatcatgca ggaatttgaa agtgacacgt ttttcccaga aattgatttg gggaaatata
     1501 aacttctccc agaataccca ggcgtcctct ctgaggtcca ggaggaaaaa ggcatcaagt
     1561 ataagtttga agtctacgag aagaaagaca cgcgtacgcg gccgctcgag cagaaactca
     1621 tctcagaaga ggatctggca gcaaatgata tcctggatta caaggatgac gacgataagg
     1681 tttaaacggc cggccgcggt catagctgtt tcctgaacag atcccgggtg gcatccctgt
     1741 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     1801 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     1861 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     1921 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     1981 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     2041 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     2101 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     2161 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta
     2221 taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga
     2281 catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct
     2341 gttgaattgg gtacgcggcc agcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3241 acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc
     3301 cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg
     3361 tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa
     3421 cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc
     3481 cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag
     3541 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     3601 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     3661 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     3721 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     3781 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcga
     3841 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     3901 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
     3961 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     4021 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4081 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4141 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     4201 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4261 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
     4321 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4381 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     4441 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     4501 catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     4561 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     4621 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     4681 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     4741 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     4801 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     4861 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
     4921 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4981 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     5041 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5101 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5161 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5221 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5281 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5341 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5401 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt tt
//
