LOCUS       40924_2662        7527 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p3FsHMGB1A-wt DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7527)
  AUTHORS   Kikuchi H, Yagi H, Hasegawa H, Ishii Y, Okabayashi K, Tsuruta M, 
            Hoshino G, Takayanagi A, Kitagawa Y.
  TITLE     Therapeutic potential of transgenic mesenchymal stem cells 
            engineered to mediate anti-high mobility group box 1 activity: 
            targeting of colon cancer
  JOURNAL   J. Surg. Res. 190 (1), 134-143 (2014)
  PUBMED    24679698
REFERENCE   2  (bases 1 to 7527)
  AUTHORS   Takayanagi A, Yagi H.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JAN-2014) Contact:Atsushi Takayanagi Keio University, 
            School of Medicine, Molecular Biology; Shinanomachi 35, Shinjuku, 
            Tokyo 160-8582, Japan
REFERENCE   3  (bases 1 to 7527)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7527)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Surg. 
            Res."; date: "2014"; volume: "190"; issue: "1"; pages: "134-143"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-JAN-2014) Contact:Atsushi Takayanagi Keio University, School of 
            Medicine, Molecular Biology; Shinanomachi 35, Shinjuku, Tokyo 
            160-8582, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(66..654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    762..810
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    821..912
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     misc_feature    927..998
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus
                     (Farrell et al., 2002)"
     misc_feature    1034..1105
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus
                     (Farrell et al., 2002)"
     enhancer        1113..1491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1524..1735
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1752..2020
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     intron          2044..2305
                     /note="bAct/bGlo hybrid Intron"
     sig_peptide     2334..2396
                     /note="enhanced human IgKappa secretion signal"
     CDS             2397..2462
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     misc_feature    2792..3380
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(3402..3523)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3615..3641
                     /label=E.coli
                     /note="E.coli"
                     /regulatory_class="terminator"
     misc_feature    complement(3747..3949)
                     /label=human TOPO1 poly(A) region
                     /note="human TOPO1 poly(A) region"
     CDS             complement(3956..4972)
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     misc_feature    complement(4999..6541)
                     /label=Intron
                     /note="Intron"
     regulatory      complement(5037..5104)
                     /label=E.coli EM2KC promoter
                     /note="E.coli EM2KC promoter"
                     /regulatory_class="promoter"
     misc_feature    5222..6198
                     /label=human TOP1 MARII
                     /note="human TOP1 MARII"
     misc_feature    6044..6198
                     /label=putative topoII binding region
                     /note="putative topoII binding region"
     misc_feature    6224..6250
                     /label=synthetic SP1-binding site
                     /note="synthetic SP1-binding site"
     regulatory      complement(6542..7523)
                     /gene="hTOP1"
                     /note="del-ATG; human TOP1"
                     /regulatory_class="promoter"
     gene            complement(6542..7523)
                     /gene="hTOP1"
                     /label=hTOP1
     misc_feature    6624
                     /label=del-Esp3I
                     /note="del-Esp3I"
ORIGIN
        1 gggcacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
       61 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      721 ttggtcatgg ctagttaatt aacatttaaa tcagcggccg caataaaata tctttatttt
      781 cattacatct gtgtgttggt tttttgtgtg aatcgtaact aacatacgct ctccatcaaa
      841 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
      901 gaacatttct ctatcgaagg cgcgcccatc ttggaccatt agctccacag gtatcttctt
      961 ccctctagtg gtcataacag cagcttcagc tacctctctc tctgctgcca gtagagggca
     1021 cacttacttt actcattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     1081 ttcatcaaac tttcttccct ccctacctgc aggcattgat tattgactag ttattaatag
     1141 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     1201 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     1261 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     1321 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     1381 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     1441 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtggatctg
     1501 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     1561 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     1621 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     1681 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     1741 gctagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc
     1801 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt
     1861 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt
     1921 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct
     1981 acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta
     2041 caggtaagtg ggcgggacgg cccttctcct tcgggctgta attagcgctt ggtttaatga
     2101 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
     2161 tgcgggggga gcggctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     2221 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     2281 tcatgccttc ttctttttcc tacagctcct gggcaacgga attcaccacc accatgaaga
     2341 aaaagctgct cgcccagctg ctcggcctgc tcatgctgtg ggtgcctagc tccggcgact
     2401 acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat gacgatgaca
     2461 agcttatggg caaaggagat cctaagaagc cgagaggcaa aatgtcatca tatgcatttt
     2521 ttgtgcaaac ttgtcgggag gagcataaga agaagcaccc agatgcttca gtcaacttct
     2581 cagagttttc taagaagtgc tcagagaggt ggaagaccat gtctgctaaa gagaaaggaa
     2641 aatttgaaga tatggcaaaa gcggacaagg cccgttatga aagagaaatg aaaacctata
     2701 tccctcccaa aggggagaca taatgacgcc cgtcacaaag agcttcaaca ggggagagtg
     2761 ttgataagct agccggttag taatgatcga caatcaacct ctggattaca aaatttgtga
     2821 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2881 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2941 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3001 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3061 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3121 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3181 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3241 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3301 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3361 cctttgggcc gcctccccgc ctggcgactc gacagacatg ataagataca ttgatgagtt
     3421 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3481 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3541 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3601 ctacaaatgt ggtaaaaaag ggcatcaaat gatgcccttt ttgcattggc tgcccaggcc
     3661 tgcactgccg cctgccggca ggggtccagt ccacgagacc cagctccctg cagtaaagta
     3721 agtgtgccct ctactggcag cagagacaac actcctttcc aggccaggcc ctgcctcatc
     3781 tgctcctggg aggaatctac tcttgctgac ctgagtggtg agacctcatc ccacactaag
     3841 tcccagcccc aactctggcc tgaggccctt tgtaacaaag aggcagacac ttaagtttcc
     3901 attaagttgt aggaatttat tttaaataac ctacagttga ttaaaagggt acctattcct
     3961 ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc
     4021 catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc
     4081 cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt
     4141 caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccgcg
     4201 gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag
     4261 ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg
     4321 cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc
     4381 cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct
     4441 catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat
     4501 acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc
     4561 cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca gcgatcgcat
     4621 ccatagcctc cgcgaccggt tgcagaacag cgggcagttc ggtttcaggc aggtcttgca
     4681 acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgcta aattccccaa
     4741 tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat
     4801 ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat
     4861 cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg gacacgctgt
     4921 cgaacttttc gatcagaaac ttctcaacag acgtcgcggt gagttcaggc tttccggatc
     4981 tatccatggt acctcctcct gtcaggagag gaaagagaag aaggttagta caattgctat
     5041 agtgagttgt attatactat gcagatatac tatgccaatg attaattgtc aaactagggc
     5101 tgcagggttc atagtgccac ttttcctgca ctgccccatc tcctgcccac cctttcccag
     5161 gcatagacag tcagtgactt accaaactca caggagggag aaggcagaag cttggccggc
     5221 caagacatgg tctcaaatga ccaacaacaa aaaccaacag aatagataag tttttatgat
     5281 ataatagaat accatgtaac aaataaaatg gattaattaa aagtgtaggt taagataata
     5341 aaaccacaaa atcaaagctg gagaggaata aacaagttgc agaatgatcc cagtaacata
     5401 cctaatttaa aactaaaacc tgtttttatt ttacaatagg ttgtacaata tgtctttagc
     5461 ttacaatagt ttacaacata ttgtaaacta aaaacaagtt ttaaattagt atgttactag
     5521 tgtgttacat attgtaaaat tacatattgt ttatgaacac ataaatatac tgcaaaaata
     5581 caaaaaatat gtatgagaat aaaaagaccg agttcatagt aatctacagg gaagaaggtg
     5641 aatggtatac aagtgacttc agctatgttt atcagtatct tttatgaaat aaaacaattg
     5701 agcacataag acataatttt aagattttac aaagccagat ggtggacata cgagtgttca
     5761 ttacataatt atctattcct gtatgtttga aatgtttcac taaaaataaa acccctaata
     5821 tttgtatcta cattttttaa atctaaaaag agaatgcttc ttccgctggc aatataaagt
     5881 taaaaattat tcaaggcact atcaaacccc aacctttgaa gatctcacta ttctgctgag
     5941 tacaagacag cctaaaagag aactgagagc aagctcaaga aatcatttct ttttctccta
     6001 atggcaccac ttttccaggc ctttaatgac tcctattggg atcccattgc agttcaagtc
     6061 aggcactttc tcctgagcat aaggttcaga gtctaactct gggtgacttt taggttcact
     6121 accatgatca tcatccagaa acttgcacaa atgtctattg tgtatagcag ttagctggat
     6181 attgtctgga ataaaattgg ccggccgaat taattcgccc gggcggcccg ccccctcggc
     6241 ccgccccctc tgcagcccct ccggcggcga gcgcgcgcgg ccggggtctg gctcaccctg
     6301 cgcccccggg gaggacgggg acccgggggt gaccacagag ggcagccatc cctgcctcga
     6361 actgtaactt cgcggccctc gctcgcggcc tagcgagccg atctactccg gcaggcggcc
     6421 cctcaccggg ggctccggcc tgtcaaagct gccgggccgg ggctggggtc ggggcctgcg
     6481 ccgggggtcg cgggacggcc aggccggggt ccggggccgc cagggtcagg ccgggccgta
     6541 cctgggaatc gttgtggagg tggtccccac tcaggtcggc ccggagggac gagcgcggcg
     6601 gcaggcgagg cggcgtgggg gagccgcaga cggcgaaccg gcctgtgcgg gcggacgggg
     6661 actccggagg ctcgaggagg cgcaggtaag cggcggtgac tgtgggcccg agactccaga
     6721 aacggctgag gctgctacca cggcggcgac gctgcctgac tgccgggcgg gcggcgagaa
     6781 cagaccccag cagttgtgta acagcctaag ttcgcatttg ggctcactgc gcctgcgcgg
     6841 ccgcttttaa acaacacgtc ggcgacgtca gcgctcgcgg aaagaggcag agggttgcgg
     6901 cccgccccct cgctgttcgc cagacggacc tgtctttgca tcgcaagccc cgcctccgga
     6961 ccgtgcgcgc tccgctgtga tttgcataag gagccgactc ctcccacggc gtcccacccc
     7021 aagcagcctc cccttcctct ccagacgctt tttctcggaa ccccagcccc tctgctaggc
     7081 agtgcgcagg cgccacccgg gcaccgcctc cggaccaggg ctggccgcgc agccacgtgg
     7141 tgggccttta aaggccgcgg cgggacccag caatcttggg cagagctggt atccggcgtc
     7201 cgcatctcct tggagcggcg agtaccttgg cagctgggac cccagaggtc tccccagcgc
     7261 ctcctttctt gctaagccga ccctgaggcc gcctggtggg aggagtcccc tccccagcct
     7321 ttgtccaggc cctccaactt ctggattatt tcacgcccac tttttaaaaa ttattttaat
     7381 ttttcaacca accaagggtc cggtttaacc ccagttttcc gccgagtaca tgtagccctg
     7441 ggtcaccaag ggtctgattt ctttggttca ttctacaaat acttattgag catccgctat
     7501 gagcccagga attgtgctga attgccc
//
