LOCUS       V029458                 6192 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029458
VERSION     V029458
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..2339
                     /label="Shh(NM_009170)"
                     /note="Shh(NM_009170)"
                     /gene="Shh"
     CDS             2358..2387
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2406..2429
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2449..2465)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2476..3098
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3247..3835)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4164..4211
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4443..5237)
                     /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(5272..5629)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5631..5735)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5762..6190
                     /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 gctgctgctg ctggccagat gttttctggt gatccttgct tcctcgctgc
     1081 tggtgtgccc cgggctggcc tgtgggcccg gcagggggtt tggaaagagg cggcacccca
     1141 aaaagctgac ccctttagcc tacaagcagt ttattcccaa cgtagccgag aagaccctag
     1201 gggccagcgg cagatatgaa gggaagatca caagaaactc cgaacgattt aaggaactca
     1261 cccccaatta caaccccgac atcatattta aggatgagga aaacacggga gcagaccggc
     1321 tgatgactca gaggtgcaaa gacaagttaa atgccttggc catctctgtg atgaaccagt
     1381 ggcctggagt gaagctgcga gtgaccgagg gctgggatga ggacggccat cattcagagg
     1441 agtctctaca ctatgagggt cgagcagtgg acatcaccac gtccgaccgg gaccgcagca
     1501 agtacggcat gctggctcgc ctggctgtgg aagcaggttt cgactgggtc tactatgaat
     1561 ccaaagctca catccactgt tctgtgaaag cagagaactc cgtggcggcc aaatccggcg
     1621 gctgtttccc gggatccgcc accgtgcacc tggagcaggg cggcaccaag ctggtgaagg
     1681 acttacgtcc cggagaccgc gtgctggcgg ctgacgacca gggccggctg ctgtacagcg
     1741 acttcctcac cttcctggac cgcgacgaag gcgccaagaa ggtcttctac gtgatcgaga
     1801 cgctggagcc gcgcgagcgc ctgctgctca ccgccgcgca cctgctcttc gtggcgccgc
     1861 acaacgactc ggggcccacg cccgggccaa gcgcgctctt tgccagccgc gtgcgccccg
     1921 ggcagcgcgt gtacgtggtg gctgaacgcg gcggggaccg ccggctgctg cccgccgcgg
     1981 tgcacagcgt gacgctgcga gaggaggagg cgggcgcgta cgcgccgctc acggcgcacg
     2041 gcaccattct catcaaccgg gtgctcgcct cgtgctacgc tgtcatcgag gagcacagct
     2101 gggcacaccg ggccttcgcg cctttccgcc tggcgcacgc gctgctggcc gcgctggcac
     2161 ccgcccgcac ggacggcggg ggcgggggca gcatccctgc agcgcaatct gcaacggaag
     2221 cgaggggcgc ggagccgact gcgggcatcc actggtactc gcagctgctc taccacattg
     2281 gcacctggct gttggacagc gagaccatgc atcccttggg aatggcggtc aagtccagca
     2341 cgcgtacgcg gccgctcgag cagaaactca tctcagaaga ggatctggca gcaaatgata
     2401 tcctggatta caaggatgac gacgataagg tttaaacggc cggccgcggt catagctgtt
     2461 tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     2521 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     2581 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     2641 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     2701 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     2761 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     2821 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     2881 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     2941 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     3001 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     3061 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcggcgagc
     3121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3661 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3961 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc
     4021 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga
     4081 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc
     4141 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct
     4201 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     4261 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     4321 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     4381 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     4441 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     4501 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     4561 cgggtagcca acgctatgtc ctgatagcga tccgccacac ccagccggcc acagtcgatg
     4621 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     4681 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc
     4741 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     4801 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     4861 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     4921 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     4981 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     5041 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     5101 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     5161 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     5221 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     5281 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     5341 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     5401 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     5461 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     5521 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5581 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     5641 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5701 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5761 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5821 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5881 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5941 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6001 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6061 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6121 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6181 aacgcgaatt tt
//
