LOCUS       40924_25566        7667 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pInSRT-GFPV3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  AUTHORS   Wasa J, Nishida Y, Shinomura T, Urakawa H, Arai E, Futamura N, 
            Tsukushi S, Ishiguro N.
  TITLE     Versican Regulates Cell-Associated Matrix Formation and Cell 
            Behavior Differentially from Aggrecan in Swarm Rat Chondrosarcoma 
            Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7667)
  AUTHORS   Shinomura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUN-2010) Contact:Tamayuki Shinomura Tokyo Medical and
            Dental University, Tissue Regeneration; 1-5-45 Yushima, Bunkyo-Ku, 
            Tokyo 113-8549, Japan
REFERENCE   3  (bases 1 to 7667)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7667)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-JUN-2010) Contact:Tamayuki Shinomura Tokyo Medical and Dental 
            University, Tissue Regeneration; 1-5-45 Yushima, Bunkyo-Ku, Tokyo 
            113-8549, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..32
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    50..97
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    177..236
                     /gene="GFPV3"
                     /label=signal sequence derived from human PG-M/versican
                     /note="signal sequence derived from human PG-M/versican"
     CDS             279..995
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3035..3621
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3622..4218
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4308..4532
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4566..4584)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4591..4607)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4748..5203)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5537..5641
                     /label=AmpR promoter
     CDS             5642..6499
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6673..7261
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7549..7570
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7585..7615
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7623..7639
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7647..7663
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attacgccaa gctatttagg tgacactata gaatactcaa gcttcgaccg aagttcctat
       61 tccgaagttc ctattctcta gaaagtatag gaacttcgga tcccctaatt caagcttgcg
      121 cagcgctgca gtgaattttc cccccaaact gcaataagcc gccttccaag gccaagatgt
      181 tcataaatat aaagagcatc ttatggatgt gttcaacctt aatagtaacc catgcgctac
      241 ataaagtcaa agtgggaaaa agcccaccgg tcgccaccat ggtgagcaag ggcgaggagc
      301 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      361 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      421 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      481 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      541 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      601 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      661 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      721 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
      781 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
      841 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
      901 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
      961 ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag
     1021 cttcgaattc tgcagtcgac ggtaccgcgg gcccggtgag gggctccctc tctggaaaag
     1081 tcagcctacc ttgtcatttt tcaacgatgc ctactttgcc acccagttac aacaccagtg
     1141 aatttctccg catcaaatgg tctaagattg aagtggacaa aaatggaaaa gatttgaaag
     1201 agactactgt ccttgtggcc caaaatggaa atatcaagat tggtcaggac tacaaaggga
     1261 gagtgtctgt gcccacacat cccgaggctg tgggcgatgc ctccctcact gtggtcaagc
     1321 tgctggcaag tgatgcgggt ctttaccgct gtgacgtcat gtacgggatt gaagacacac
     1381 aagacacggt gtcactgact gtggatgggg ttgtgtttca ctacagggcg gcaaccagca
     1441 ggtacacact gaattttgag gctgctcaga aggcttgttt ggacgttggg gcagtcatag
     1501 caactccaga gcagctcttt gctgcctatg aagatggatt tgagcagtgt gacgcaggct
     1561 ggctggctga tcagactgtc agatatccca tccgggctcc cagagtaggc tgttatggag
     1621 ataagatggg aaaggcagga gtcaggactt atggattccg ttctccccag gaaacttacg
     1681 atgtgtattg ttatgtggat catctggatg gtgatgtgtt ccacctcact gtccccagta
     1741 aattcacctt cgaggaggct gcaaaagagt gtgaaaacca ggctgccagg ctggcaacag
     1801 tgggggaact ccaggcggca tggaggaacg gctttgacca gtgcgattac gggtggctgt
     1861 cggatgccag cgtgcgccac cctgtgactg tggccagggc ccagtgtgga ggtggtctac
     1921 ttggggtgag aaccctgtat cgttttgaga accagacagg cttccctccc cctgatagca
     1981 gatttgatgc ctactgcttt aaacgacctg atcgctgcaa aatgaacccg tgccttaacg
     2041 gaggcacctg ttatcctact gaaacttcct acgtatgcac ctgtgtgcca ggatacagcg
     2101 gagaccagtg tgaacttgat tttgatgaat gtcactctaa tccctgtcgt aatggagcca
     2161 cttgtgttga tggttttaac acattcaggt gcctctgcct tccaagttat gttggtgcac
     2221 tttgtgagca agataccgag acatgtgact atggctggca caaattccaa gggcagtgct
     2281 acaaatactt tgcccatcga cgcacatggg atgcagctga acgggaatgc cgtctgcagg
     2341 gtgcccatct cacaagcatc ctgtctcacg aagaacaaat gtttgttaat cgtgtgggcc
     2401 atgattatca gtggataggc ctcaatgaca agatgtttga gcatgacttc cgttggactg
     2461 atggcagcac actgcaatac gagaattgga gacccaacca gccagacagc ttcttttctg
     2521 ctggagaaga ctgtgttgta atcatttggc atgagaatgg ccagtggaat gatgttccct
     2581 gcaattacca tctcacctat acgtgcaaga aaggaacagt tgcttgcggc cagccccctg
     2641 ttgtagaaaa tgccaagacc tttggaaaga tgaaacctcg ttatgaaatc aactccctga
     2701 ttagatacca ctgcaaagat ggtttcattc aacgtcacct tccaactatc cggtgcttag
     2761 gaaatggaag atgggctata cctaaaatta cctgcatgaa cccatctgca taccaaagga
     2821 cttattctat gaaatacttt aaaaattcct catcagcaaa ggacaattca ataaatacat
     2881 ccaaacatga tcatcgttgg agccggaggt ggcaggagtc gaggcgctga tccctaaaat
     2941 ggcgaacatg tgttttcatc atttcagcca aagtcctaac ttcctgtgcc tttcctatca
     3001 cctcgacaag cttgggctgc aggtccgccc cccccccctc tccctccccc ccccctaacg
     3061 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3121 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3181 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3241 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     3301 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     3361 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     3421 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     3481 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     3541 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     3601 gatgataata tggccacaac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3661 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3721 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3781 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3841 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     3901 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     3961 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4021 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     4081 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4141 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4201 acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     4261 gcacgacccc atgcatcgat gatctagagc tcgctgatca gcctcgactg tgccttctag
     4321 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4381 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4441 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4501 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa cctgcagcaa
     4561 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     4621 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4681 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4741 cgaatggaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaatat ttgttaaatc
     4801 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4861 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4921 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4981 tcacccaaat caagtttttt gcggtcgagg tgccgtaaag ctctaaatcg gaaccctaaa
     5041 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5101 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5161 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcgcctga tgcggtattt
     5221 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     5281 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5341 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5401 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5461 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5521 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5581 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5641 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5701 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5761 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5821 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtcata cactattatc
     5881 ccgtattgac gccgggcaag agcaactcgg tcgccgggcg cggtattctc agaatgactt
     5941 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6001 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6061 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6121 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6181 gcctgtagca atgccaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6241 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6301 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6361 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6421 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6481 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6541 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6601 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6661 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6721 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6781 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     6841 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6901 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6961 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7021 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7081 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7141 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7201 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatcgaa
     7261 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7321 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     7381 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     7441 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     7501 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     7561 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     7621 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatg
//
