LOCUS       V030318                 5382 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030318
VERSION     V030318
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..1529
                     /label="Hes5(NM_010419)"
                     /note="Hes5(NM_010419)"
                     /gene="Hes5"
     CDS             1548..1577
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             1596..1619
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(1639..1655)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    1666..2288
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(2437..3025)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3354..3401
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(3633..4427)
                     /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(4462..4819)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(4821..4925)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      4952..5380
                     /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 ggccccaagt accgtggcgg tggagatgct cagtcccaag gagaaaaacc
     1081 gactgcggaa gccggtggtg gagaagatgc gtcgggaccg catcaacagc agcatagagc
     1141 agctgaagct gctgctggag caggagttcg cgcggcacca gcccaactcc aagctggaga
     1201 aggccgacat cctggagatg gccgtcagct acctgaaaca cagcaaagcc ttcgccgcgg
     1261 ccgccggccc caagagcctg caccaggact acagcgaggg ctactcctgg tgcctgcagg
     1321 aggcggtaca gttcctgacc ctgcacgccg ccagcgacac gcagatgaag ctgctttacc
     1381 acttccagcg gcccccagct cccgccgctc ctgccaagga gcccccggcg cccggagctg
     1441 cgccccaacc ggcccgctcc tccgctaagg ctgctgctgc tgcggtctcc acctcgcgcc
     1501 aacccgcctg cggcctctgg cggccctgga cgcgtacgcg gccgctcgag cagaaactca
     1561 tctcagaaga ggatctggca gcaaatgata tcctggatta caaggatgac gacgataagg
     1621 tttaaacggc cggccgcggt catagctgtt tcctgaacag atcccgggtg gcatccctgt
     1681 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     1741 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     1801 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     1861 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     1921 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     1981 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     2041 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     2101 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta
     2161 taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga
     2221 catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct
     2281 gttgaattgg gtacgcggcc agcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2341 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2941 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3001 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3061 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3121 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3181 acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc
     3241 cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg
     3301 tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa
     3361 cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc
     3421 cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag
     3481 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     3541 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     3601 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     3661 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     3721 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcga
     3781 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     3841 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
     3901 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     3961 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4021 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4081 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     4141 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4201 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
     4261 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4321 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     4381 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     4441 catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     4501 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     4561 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     4621 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     4681 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     4741 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     4801 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
     4861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4921 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     4981 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5041 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5101 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5161 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5221 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5281 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5341 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt tt
//
