LOCUS       40924_36408        4899 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector prCol2(P)-FRT DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4899)
  AUTHORS   Ito K, Shinomura T.
  TITLE     Development and application of a new Silent reporter system to 
            quantitate the activity of enhancer elements in the type II Collagen
            Gene
  JOURNAL   Gene (2016) In press
  PUBMED    26992640
REFERENCE   2  (bases 1 to 4899)
  AUTHORS   Shinomura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-OCT-2015) 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 4899)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4899)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene (2016)
            In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-OCT-2015) 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..4899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..32
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     gene            78..1727
                     /gene="Col2a1"
                     /label=Col2a1
     regulatory      78..1066
                     /gene="Col2a1"
                     /label=rat type II collagen
                     /note="rat type II collagen"
                     /regulatory_class="promoter"
     exon            1067..1307
                     /gene="Col2a1"
                     /number=1
                     /note="rat type II collagen"
     intron          1308..1351
                     /gene="Col2a1"
                     /number=1
                     /note="rat type II collagen partial intron"
     intron          1556..1710
                     /gene="Col2a1"
                     /number=2
                     /note="rat type II collagen partial intron"
     exon            1711..1727
                     /gene="Col2a1"
                     /number=3
                     /note="rat type II collagen partial exon"
     protein_bind    1736..1783
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(1798..1816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1823..1839)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1980..2435)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2769..2873
                     /label=AmpR promoter
     CDS             2874..3731
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3905..4493
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4781..4802
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4817..4847
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4855..4871
                     /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     4879..4895
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attacgccaa gctatttagg tgacactata gaatactcaa gctagtcaat tgtacagatc
       61 tgctagctcg agaattcaac taggaaactc tggcgctttc tcctcccctc acaaaactga
      121 gtccagctgg agccacctcc agactctctg gccagggcct cagagtggtc aacagtccct
      181 ggccagcgtt gctctctcca ggctaagggc acccactccc ctggagattc ctgaacctgg
      241 gccaggaaga gccgaattag acaagtgtct ccaatccggc tgcgtgcgga ttttgttgcg
      301 gtgtccctcg gttgtctgca gttcctttag tcccttccct ggcctgcccc ttacacctcc
      361 acacaggtcc ccctctgtgt aggaatacac cagaccctct cttagccaca cacacctcca
      421 gtcccccgtc tacctagatt tttttcatag ctagttggat gggggatggg ttagggaggc
      481 tgggtttgcg agcctccagg tgggagttca ccgacaggta ctccgcaaag gagctggaag
      541 gcaggtctgg aaaactgtcc cccagattta ggattctggg cagctttcat cagcttatac
      601 tttggctccc ccgcccccta aactccccat ccccaccttc ctttctcccg ttacttcgtc
      661 ctccctcgcc tttccagcct tgagtctaaa gctccatgct tatgcctctg caaacaaccc
      721 cctcccttct aaccccagca gaactccgag gaaaggggcc ggaggccccc cttctcgcct
      781 gtggttagag ggggcagtgt ggcagtccca agtgggggcg accggaggcc gtctcggtgc
      841 cccgcccgat caggccactg ggcacatcgg gggcgggaag ctgggctcac caaaggggcg
      901 actggccttg gcaggtgtgg gctctggtcc ggcctgggcg ggctccgggg gcggggtctc
      961 aggttacagc cccgcggggg gctggggggc ggcccgcggt ttgggctggt ttgccagcct
     1021 ttggagcgac cgggagcata taaccggagc ctctgctggg agaagacgca gagcgccgct
     1081 gggctgccgg gtctcctgcc tcctcctcct gctcctagag cctcctgcat gagggcgcgg
     1141 tagagacccg gacccgctcc gtgctctgcc gcctcgccga gcttcgcccg ggccaaggct
     1201 ctgccaggcc tcgcggtgag ccctgatccg cctcggggct ccccagtcgc tggtgctgct
     1261 gacgctgctc atcgccacgg tcctacaatg tcagggccag gatgcccgta agtcgcccac
     1321 cacccctgcc tgcttccctg acttgcgacc cttctcttct tccctccgtc cgagttaggg
     1381 cgccaagtcc taggcgcgta gtgcacagga gaacactgat cctaatccta attctgctag
     1441 tgaggagttc tgtcgcagca tcctcagtca gagtcgcaag tggagacatt ctcaggtccc
     1501 tgcgttaagc aagggtatga acttgcagag tgtcgacgcg taccggtcgc gactagtgat
     1561 cctaacccca ccacagaggg attccgggga cccacagggg tcttcttcct tccttggttg
     1621 tggtcctctc ctttccgatc ctccaaagag agtttgaatg gcaaaatatc gtctctgtct
     1681 caccttttcc ttctctgttt tcccttgcag gaaaattagg gccaaagctt cgaccgaagt
     1741 tcctattccg aagttcctat tctctagaaa gtataggaac ttcggatcgc aattcgccct
     1801 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     1861 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     1921 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     1981 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat atttgttaaa tcagctcatt
     2041 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2101 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2161 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
     2221 atcaagtttt ttgcggtcga ggtgccgtaa agctctaaat cggaacccta aagggagccc
     2281 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2341 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2401 acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcct gatgcggtat tttctcctta
     2461 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     2521 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2581 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2641 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2701 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2761 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2821 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2881 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2941 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3001 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3061 gttttccaat gatgagcact tttaaagttc tgctatgtca tacactatta tcccgtattg
     3121 acgccgggca agagcaactc ggtcgccggg cgcggtattc tcagaatgac ttggttgagt
     3181 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3241 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3301 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3361 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3421 caatgccaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3481 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3541 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3601 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3661 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3721 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3781 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3841 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3901 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3961 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4021 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4081 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4141 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4201 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4261 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4321 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4381 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4441 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatcg aaaaacgcca
     4501 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4561 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4621 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4681 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4741 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     4801 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     4861 gcggataaca atttcacaca ggaaacagct atgaccatg
//
