LOCUS       V031232                 7665 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031232
VERSION     V031232
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..3812
                     /label="Gprc6a(NM_153071)"
                     /note="Gprc6a(NM_153071)"
                     /gene="Gprc6a"
     CDS             3831..3860
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3879..3902
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3922..3938)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3949..4571
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4720..5308)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5637..5684
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5916..6710)
                     /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(6745..7102)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7104..7208)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7235..7663
                     /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 ggccctattg attacagtag ttacttgctt catgattatt cttgatacct
     1081 cccagtcttg tcatacccca gatgacttcg tggctatcac ttctcctgga catatcatga
     1141 ttggtggctt gtttgccatt catgaaaaaa tgttgtcctc agatgaccat cccagacgac
     1201 cacaaatcca gaaatgtgct ggctttgaaa tatcagtgtt tcttcaaact ctggctatga
     1261 tacacagcat tgagatgatc aataattcaa cactgttatc cggagtcaag ctgggatatg
     1321 aaatctatga cacttgtacc gaagtcacag cagcaatggc tgccactctg aggttcctct
     1381 caaaattcaa ctgctctaga gaaactgtgg tctttcagtg tgactattcc agctacatgc
     1441 caagggtcaa ggctgtcata ggtgctggct actctgaaac atccatcgcg gtctcaagga
     1501 tgctgaactt acagctcatg ccacaggtga gttatgaatc cactgcagaa atcctgagtg
     1561 acaagatccg ctttccttca tttttacgga ctgtgcctag tgacttctac caaactaaag
     1621 caatggccca cctgatccga caatcgggat ggaactggat tggtgccata acaaccgatg
     1681 atgattatgg aagactagct ctcaacacat ttgcaataca ggctgctgaa aacaatgtgt
     1741 gcattgcctt caaagaggtt ctgccagcct tcctctcaga taataccatt gaagtgagaa
     1801 ttaaccagac tctggagaag atcattgcag aagcccaggt taatgtcatt gtggtgtttc
     1861 tgagaaaatt ccatgttttc aatctcttca cgaaagccat tgaaaggaaa ataagtaaga
     1921 tctggattgc tagtgataat tggtcaaccg ctaccaagat tatcaccatc cctaatgtta
     1981 agaagcttgg caaagtggtg gggtttgcct ttaggagagg aaatacgtct tctttccatt
     2041 cttttcttca aactctgcat atgtacccca atgacaataa caaaccccta catgaatttg
     2101 ccatgctcgt ttctgcctgt aaatacatca aagacggtga tttgagccaa tgcatatcca
     2161 actattctca ggcaactttg acctatgaca ctaccaagac cattgaaaac cacttattca
     2221 agagaaatga tttcctctgg cattatactg agccgggact catttatagc attcagcttg
     2281 cagtgttcgc ccttggtcat gccatccggg atctgtgcca agctcgagac tgcaagaaac
     2341 ccaacgcctt tcagccatgg gagctacttg ccgtgctgaa aaacgtgaca ttcactgatg
     2401 gaaggaattc atttcacttt gatgcccacg gggatttaaa tactggttat gacgtggtgc
     2461 tctggaagga gaccaatggc ctcatgactg tcacaaagat ggcagaatat gacctgcagc
     2521 gtgacgtctt catcaccaca aaccaagaaa caaaacatga attcaggaaa cttaagcaaa
     2581 ttctatctaa atgctccaag gaatgcagcc ctggtcaaat gaagaaagcc acaggaagcc
     2641 aacacagctg ttgctatgaa tgtgtgagct gcccagaaaa ccactacagt aatgagacag
     2701 atatggatca ctgcctttta tgcaacaatg aaacccactg ggccccagta aggagcacta
     2761 cctgctttga aaaggaagtg gagtatctgg actgggatga ctccttggct ctcctcctca
     2821 ttgccctctc cctacttgga atcgcctttg ttctggccat tggcataata tttacaagaa
     2881 acctgaagac acctgtcgtg aaatcatctg gaggattagt ggtctgctac gtgatgctca
     2941 tctgccacgc ccttaacttt gccagcacag gcttttttat tggagaacca caagactttg
     3001 cgtgcaagac caggcagacc ttatttggtg tgagctttac tctctgtgtc tcctgtattt
     3061 tgaccaagtc cctgaaaatt ttgctagctt tcagttttga ccccaagctg acaatgttcc
     3121 tgaagtgcct ttataggcct gttcccattg ttcttacctg cacgggcatt caagtggtca
     3181 tttgcactct ttggctggtg ctggcagcac ctagtgtgga agagaatatc tccttgccta
     3241 gagtcattat cctggaatgt gaggaggggt cagcactggc atttggtacc atgctgggct
     3301 acatcacagt tctggccttc atttgctttg tatttgcatt caaggggagg aaacttcctg
     3361 agaattacaa cgaagccaag ttcctgacct ttgggatgct gatttacttc atagcttgga
     3421 tcacattcat ccctgtctat actaccacat tcggcaagta tttgcccgcc gtggagatta
     3481 tagtcattct gatatccaat tatgggattc tctgctgtat attcttcccc aagtgctaca
     3541 ttattctttg caagcagaag actaacacca aatcagcctt cctccagatg gtttacaact
     3601 actctgctca cagtgtggac agccttgcct tgagtcatgt ttccctggac tccaccagct
     3661 acgatactgc aacaaccaat cagagccctg gtaacaagat gacagcctgt cagaacgaca
     3721 accatcttcc tgcacaagtg ctcccacaca caggcacggc aaagactatc aaagcgtcta
     3781 aaactttgcg tcagaaaaga agttcaagta taacgcgtac gcggccgctc gagcagaaac
     3841 tcatctcaga agaggatctg gcagcaaatg atatcctgga ttacaaggat gacgacgata
     3901 aggtttaaac ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc
     3961 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     4021 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     4081 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     4141 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     4201 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     4261 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     4321 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     4381 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     4441 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     4501 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     4561 cctgttgaat tgggtacgcg gccagcggcg agcggtatca gctcactcaa aggcggtaat
     4621 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5101 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5281 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5341 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5401 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5461 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     5521 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     5581 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     5641 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     5701 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     5761 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     5821 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     5881 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     5941 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     6001 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     6061 cgatccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     6121 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     6181 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     6241 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     6301 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     6361 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     6421 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     6481 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     6541 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     6601 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     6661 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     6721 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     6781 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     6841 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     6901 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     6961 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     7021 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     7081 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
     7141 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7201 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     7261 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     7321 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     7381 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     7441 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     7501 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     7561 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     7621 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga atttt
//
