LOCUS       V030360                 6264 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030360
VERSION     V030360
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..2411
                     /label="Porcn(NM_023638)"
                     /note="Porcn(NM_023638)"
                     /gene="Porcn"
     CDS             2430..2459
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2478..2501
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2521..2537)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2548..3170
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3319..3907)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4236..4283
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4515..5309)
                     /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(5344..5701)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5703..5807)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5834..6262
                     /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 ggccaccttc agccgccagg aatttttcca gcagctactg cagggctgtc
     1081 tcctgcctac tgtccaacag ggccttgacc agatctggct gcttcttacc atctgcttcg
     1141 cctgccgcct cctttggagg ctggggttac cgtcttacct gaagcatgca agcaccgtgg
     1201 caggtggctt cttcagcctc taccacttct tccagctgca catggtttgg gtcgtgctgc
     1261 tgagcctcct gtgctacctc gtgctgttcc tctgccgaca ctcctcccac cgaggcgtct
     1321 tcctctccgt caccatcctc atctacctgc tcatgggtga gatgcacatg gtggacaccg
     1381 tgacatggca caagatgcga ggggcccaga tgatcgtggc catgaaggcg gtgtctctgg
     1441 gcttcgacct ggaccggggc gaggtgggtg cagtgccctc acctgtggag ttcatgggct
     1501 acctctactt tgtgggcacc atcgtctttg ggccctggat atccttccac agctacctac
     1561 aggctgtcca aggccgccca ctgagccgcc gatggctgaa gaaggtggcc cggagcctgg
     1621 cgctggccct gctgtgcctt gtactgtcca cttgtgtggg cccctacctc ttcccctact
     1681 tcatccccct cgacggtgac cgactccttc gcaacaagaa acgcaaagcc aggggcacca
     1741 tggtaaggtg gctacgagcc tacgagagtg ctgtctcctt ccacttcagc aactattttg
     1801 tgggctttct gtctgaggcc acagccacat tggccggggc tggcttcacg gaggagaagg
     1861 accacctgga atgggacctg acagtctcta gaccgttgaa tgtggagctg ccccggtcca
     1921 tggtggaagt tgtcacaagc tggaacctgc ccatgtctta ttggttaaat aattatgttt
     1981 tcaaaaatgc ccttcgcctg gggaccttct ctgctgtgct ggtcacctat gcagccagtg
     2041 ccctcctgca tggcttcagt ttccacctgg ctgctgtgct gctgtccctg gcattcatca
     2101 cttatgtgga acatgtcctc agaaagcgcc tggctcagat cctcagtgcc tgcatcttgt
     2161 caaagcgttg tctgccagac tgttcacatc ggcatcgctt gggcctgggg gtacgagcct
     2221 taaacttgct ctttggggcc ctggctatct tccacctgtc ctacctgggc tccctgtttg
     2281 atgtcgatgt ggacgacacc acagaggagc agggctacgg catggcatac actgtccaca
     2341 agtggtcaga gctcagctgg gccagtcact gggtcacttt tggatgctgg atcttctacc
     2401 gtctcatagg cacgcgtacg cggccgctcg agcagaaact catctcagaa gaggatctgg
     2461 cagcaaatga tatcctggat tacaaggatg acgacgataa ggtttaaacg gccggccgcg
     2521 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct
     2581 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     2641 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     2701 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     2761 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     2821 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     2881 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     2941 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3001 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     3061 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     3121 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     3181 ccagcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3721 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4021 ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg ctatggcagg
     4081 gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg gggggtgggg
     4141 tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg gggtatcgac
     4201 agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt
     4261 tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg
     4321 tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat ccccgcgctg
     4381 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     4441 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     4501 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     4561 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     4621 tcagcaatat cacgggtagc caacgctatg tcctgatagc gatccgccac acccagccgg
     4681 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     4741 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac
     4801 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     4861 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     4921 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     4981 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     5041 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     5101 agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc
     5161 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     5221 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     5281 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg
     5341 atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     5401 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     5461 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5521 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5581 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5641 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     5701 ggactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5761 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5821 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5881 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5941 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6001 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6061 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6121 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6181 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6241 taacaaaaat ttaacgcgaa tttt
//
