LOCUS       V031079                 5685 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031079
VERSION     V031079
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..1832
                     /label="Fgf8(NM_010205)"
                     /note="Fgf8(NM_010205)"
                     /gene="Fgf8"
     CDS             1851..1880
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             1899..1922
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(1942..1958)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    1969..2591
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(2740..3328)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    3657..3704
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(3936..4730)
                     /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(4765..5122)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5124..5228)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5255..5683
                     /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 gggcagcccc cgctccgcgc tgagctgcct gctgttgcac ttgctggttc
     1081 tctgcctcca agcccaggta aggagcgctg cgcagaagcg ggggccgggc gcggggaacc
     1141 cagctgacac tctcgggcag ggacacgagg accgaccctt cggccagcgc agcagggctg
     1201 gaaagaactt tacaaatcca gccccaaact accccgagga gggatctaag gaacagagag
     1261 acagtgtcct gcctaaagtc acacagcgac atgtgaggga gcagagcctg gtgacggatc
     1321 agctcagccg ccgcctcatc cggacctacc agctctacag ccgcaccagc gggaagcacg
     1381 tgcaggtcct ggccaacaag cgcatcaacg ccatggcaga agacggagac cccttcgcga
     1441 agctcattgt ggagaccgat acttttggaa gcagagtccg agttcgcggc gcagagacag
     1501 gtctctacat ctgcatgaac aagaagggga agctaattgc caagagcaac ggcaaaggca
     1561 aggactgcgt attcacagag atcgtgctgg agaacaacta cacggcgctg cagaacgcca
     1621 agtacgaggg ctggtacatg gcctttaccc gcaagggccg gccccgcaag ggctccaaga
     1681 cgcgccagca tcagcgcgag gtgcacttca tgaagcgcct gccgcggggc caccacacca
     1741 ccgagcagag cctgcgcttc gagttcctca actacccgcc cttcacgcgc agcctgcgcg
     1801 gcagccagag gacttgggcc ccggagcccc gaacgcgtac gcggccgctc gagcagaaac
     1861 tcatctcaga agaggatctg gcagcaaatg atatcctgga ttacaaggat gacgacgata
     1921 aggtttaaac ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc
     1981 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2041 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     2101 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     2161 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     2221 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     2281 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     2341 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     2401 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     2461 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     2521 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     2581 cctgttgaat tgggtacgcg gccagcggcg agcggtatca gctcactcaa aggcggtaat
     2641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3181 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3481 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     3541 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     3601 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     3661 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     3721 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     3781 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     3841 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     3901 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     3961 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     4021 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     4081 cgatccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     4141 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     4201 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     4261 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     4321 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     4381 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     4441 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     4501 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     4561 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     4621 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     4681 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     4741 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     4801 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     4861 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     4921 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     4981 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     5041 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     5101 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
     5161 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5221 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     5281 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5341 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5401 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5461 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5521 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5581 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5641 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga atttt
//
