LOCUS       V031055                 9240 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031055
VERSION     V031055
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..5387
                     /label="Col1a1(NM_007742)"
                     /note="Col1a1(NM_007742)"
                     /gene="Col1a1"
     CDS             5406..5435
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             5454..5477
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(5497..5513)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    5524..6146
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(6295..6883)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    7212..7259
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(7491..8285)
                     /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(8320..8677)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(8679..8783)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8810..9238
                     /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 gttcagcttt gtggacctcc ggctcctgct cctcttaggg gccactgccc
     1081 tcctgacgca tggccaagaa gacatccctg aagtcagctg catacacaat ggcctaaggg
     1141 tccccaatgg tgagacgtgg aaacccgagg tatgcttgat ctgtatctgc cacaatggca
     1201 cggctgtgtg cgatgacgtg caatgcaatg aagaactgga ctgtcccaac ccccaaagac
     1261 gggagggcga gtgctgtgct ttctgcccgg aagaatacgt atcaccaaac tcagaagatg
     1321 taggagtcga gggacccaag ggagaccctg gcccccaagg cccaagggga cccgttggcc
     1381 cccctggacg agatggcatc cctggacagc ctggacttcc tggtcctcct ggtccccctg
     1441 ggccccccgg accccctggc cttggaggaa actttgcttc ccagatgtcc tatggctatg
     1501 atgaaaaatc agctggagtt tccgtgcctg gccccatggg tccttctggt cctcgtggtc
     1561 tccctggccc ccctggtgca cctggtccac aaggtttcca aggcccccct ggtgaacctg
     1621 gcgagcctgg cggttcaggt ccaatgggtc cccgaggtcc ccctggccct cctggcaaga
     1681 atggagatga tggggaagct ggcaagcccg gccgtcctgg tgagcgtgga cctcctggac
     1741 ctcagggtgc tcgtggattg cctggaacag ctggcctccc tggaatgaag ggacaccgag
     1801 gcttcagtgg tttggatggt gccaaaggag atgctggtcc tgctggtcct aagggagagc
     1861 ccggcagtcc tggtgaaaac ggagctcctg gccagatggg tccccgaggt ctgcccggtg
     1921 agagaggtcg ccctggacct cctggcactg ctggtgctcg cggtaacgat ggtgctgttg
     1981 gtgctgctgg accccctggt cccaccggcc ccactggccc tcctggcttc cctggtgcag
     2041 ttggtgctaa gggtgaagct ggtccccaag gagctagagg ctctgaaggt ccccagggtg
     2101 tgcgtggtga gcccggaccc cctggccctg ctggtgctgc cggccctgct ggaaaccctg
     2161 gtgctgatgg acaacctggc gctaaaggtg ccaatggtgc tcctggtatt gctggtgctc
     2221 ctggcttccc tggtgcccga ggcccctctg gaccccaggg ccccagcggc cctccaggtc
     2281 ccaagggtaa cagtggtgaa cctggtgctc ctggcaacaa aggagacact ggtgccaaag
     2341 gagaacccgg tgctactgga gttcaaggtc ccccaggccc tgccggagaa gaaggaaaac
     2401 gaggagcccg tggtgagcct ggaccttccg gactgcctgg acctcctggc gagcgtggtg
     2461 gacctggtag ccgtggtttc cctggtgctg atggtgttgc tggccccaag ggtccttccg
     2521 gtgaacgtgg tgctcccgga cctgctggtc ccaaaggttc tcctggtgaa gctggtcgcc
     2581 ccggtgaagc tggtctccct ggtgccaagg gtctcactgg cagtcctggc agccctggtc
     2641 ctgatggcaa aaccggcccc cctggtcccg ctggtcaaga tggtcgccct ggacccgcag
     2701 gtcctcctgg agcccgtggc caggctggtg tgatgggatt ccctggacct aagggtaccg
     2761 ctggagaacc tggaaaggct ggagagcgag gccttcccgg accccctggc gctgttggtc
     2821 ctgctggcaa agatggagaa gctggagctc agggagcccc tggccctgct ggtcctgctg
     2881 gtgagagagg tgaacaaggt cccgctggct cccctggatt ccagggtctt cctggtcctg
     2941 ccggtcctcc tggtgaagca ggcaagcctg gtgaacaggg tgttcctgga gaccttggtg
     3001 cccctggacc ctctggcgca agaggcgaga gaggtttccc tggtgaacgt ggtgtacaag
     3061 gtcccccagg tcctgctggt ccccgaggaa acaatggtgc ccccggcaac gatggtgcca
     3121 agggtgatac tggtgccccc ggagctcccg gtagccaggg tgcccccggt cttcagggaa
     3181 tgcctggtga acgtggtgca gctggtcttc caggtcctaa gggtgacaga ggtgatgctg
     3241 gtcccaaagg tgctgatggt tctcctggta aagatggtgc ccgtggtctg actggtccca
     3301 ttggtcctcc tggccctgct ggtgcccctg gtgacaaggg tgaagctggt cccagtggtc
     3361 ctcccggtcc caccggagcc cgtggtgctc ccggagaccg tggtgaggct ggtccccctg
     3421 gtcctgctgg ctttgccggc ccccctggtg ctgatggcca acctggtgcg aaaggtgaac
     3481 ctggtgatac tggtgttaaa ggtgatgctg gtcctcctgg ccctgctggt cctgctggac
     3541 cccccggccc cattggtaac gttggtgctc ctggacccaa aggtcctcgt ggtgctgctg
     3601 gtccccctgg tgctactggc ttccctggtg ctgctggccg tgtcggtccc cctggtccct
     3661 ctggaaatgc tggaccccct ggccctcccg gtcccgttgg caaagaaggg ggcaaaggtc
     3721 cccgtggtga gactggccct gctggacgtc ctggtgaagt tggtccccca ggtccccccg
     3781 gtcctgctgg tgagaaagga tctcctggtg ctgatggacc tgctggctct cctggtaccc
     3841 ctggacctca gggtattgct ggacaacgtg gtgtggtcgg tcttcccggt cagagaggag
     3901 aaagaggctt ccctggtctt cctggcccct ctggtgaacc tggcaaacaa ggtccttctg
     3961 gatcaagtgg tgaacgcggt ccccctggcc ccatggggcc ccctggattg gctggtcccc
     4021 ctggtgaatc tggacgtgag ggatcccctg gtgctgaagg ctcccctgga agggatggtg
     4081 ctcccggggc caagggtgac cgtggtgaga ctggccccgc tggcccccct ggtgcccctg
     4141 gtgctcccgg tgctcccggc cctgttggtc ccgctggcaa gaatggcgat cgtggtgaga
     4201 ctggtcctgc tggtcctgct ggtcccattg gccctgctgg tgcccgtggc cctgctggac
     4261 cccaaggccc ccgtggtgac aagggtgaga caggcgaaca aggtgacaga ggcataaagg
     4321 gtcatcgtgg cttctctggt ctccagggtc ctcctggttc tcctggttct cctggtgaac
     4381 aaggcccctc tggagcttca ggtcctgcag gcccccgggg tccccctggc tctgctggtt
     4441 ctcctggcaa agacggactc aacggtctcc ctggccccat tggtccccct ggtcctcgag
     4501 gtcgcactgg tgacagcggc cctgctggtc cccccggccc tcctggaccc cctggccctc
     4561 ctggacctcc cagtggcggt tatgacttca gcttcctgcc tcagccacct caagagaagt
     4621 ctcaagatgg tggccgctac taccgggccg atgatgctaa cgtggttcgt gaccgtgacc
     4681 ttgaggtgga caccaccctc aagagcctga gtcagcagat tgagaacatc cgcagccccg
     4741 aaggcagccg caagaaccct gcccgcacat gccgcgacct caagatgtgc cactctgact
     4801 ggaagagcgg agagtactgg atcgacccta accaaggctg caacctggac gccatcaagg
     4861 tctactgcaa catggagaca ggtcagacct gtgtgttccc tactcagccg tctgtgcctc
     4921 agaagaactg gtacatcagc ccgaacccca aggaaaagaa gcacgtctgg tttggagaga
     4981 gcatgaccga tggattcccg ttcgagtacg gaagcgaggg ctccgacccc gccgatgtcg
     5041 ctatccagct gaccttcctg cgcctaatgt ccaccgaggc ctcccagaac atcacctatc
     5101 actgcaagaa cagcgtagcc tacatggacc agcagactgg caacctcaag aaggccctgc
     5161 tcctccaggg atccaacgag atcgagctca gaggcgaagg caacagtcgc ttcacctaca
     5221 gcacccttgt ggacggctgc acgagtcaca ccggaacttg gggcaagaca gtcatcgaat
     5281 acaaaaccac caagacctcc cgcctgccca tcatcgatgt ggctcccttg gacattggtg
     5341 ccccagacca ggaattcgga ctagacattg gccctgcctg cttcgtgacg cgtacgcggc
     5401 cgctcgagca gaaactcatc tcagaagagg atctggcagc aaatgatatc ctggattaca
     5461 aggatgacga cgataaggtt taaacggccg gccgcggtca tagctgtttc ctgaacagat
     5521 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     5581 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     5641 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     5701 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     5761 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     5821 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     5881 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     5941 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     6001 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc
     6061 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg
     6121 tccactcagt agatgcctgt tgaattgggt acgcggccag cggcgagcgg tatcagctca
     6181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6481 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6721 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7021 ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc
     7081 gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc
     7141 gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     7201 cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt
     7261 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg
     7321 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     7381 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     7441 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     7501 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     7561 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     7621 gctatgtcct gatagcgatc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     7681 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     7741 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     7801 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     7861 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     7921 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     7981 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     8041 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     8101 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     8161 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     8221 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     8281 atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc
     8341 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     8401 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     8461 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     8521 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     8581 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     8641 ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat
     8701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     8821 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     8881 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     8941 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     9001 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     9061 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     9121 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     9181 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
//
