LOCUS       Exported                7527 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p3FsHMGB1A-wt DNA, complete sequence.
ACCESSION   AB902850
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7527
                     /organism="Cloning vector p3FsHMGB1A-wt"
                     /mol_type="other DNA"
                     /db_xref="taxon:1451150"
     rep_origin      5..734
                     /label=mini-pUC ori
                     /note="mini-pUC ori"
     rep_origin      complement(66..654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    762..918
                     /label=synthetic poly(A) site
                     /note="synthetic poly(A) site"
     polyA_signal    762..810
                     /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        1112..1491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      1114..1493
                     /regulatory_class="enhancer"
                     /gene="CMV"
     gene            1114..1493
                     /gene="CMV"
                     /label=CMV
     regulatory      1506..1735
                     /regulatory_class="promoter"
                     /gene="hE2F1a"
                     /note="human E2F1a"
     gene            1506..1735
                     /gene="hE2F1a"
                     /label=hE2F1a
     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"
     misc_feature    1754..2041
                     /label=HTLV R-U5
                     /note="HTLV R-U5"
     intron          2044..2305
                     /note="bAct/bGlo hybrid Intron"
     CDS             2334..2723
                     /codon_start=1
                     /product="3xFLAG-hHMGB1 Abox"
                     /label=3xFLAG-hHMGB1 Abox
                     /protein_id="BAO48052.1"
                     /translation="MKKKLLAQLLGLLMLWVPSSGDYKDHDGDYKDHDIDYKDDDDKLM
                     GKGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKKCSERWKTMSAKEKGKF
                     EDMAKADKARYEREMKTYIPPKGET"
     sig_peptide     2334..2396
                     /note="enhanced human IgKappa secretion signal"
     CDS             2397..2462
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2787..3384
                     /label=WPRE
                     /note="WPRE"
     misc_feature    2792..3380
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3263..3274)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3393..3614
                     /label=SV40 late poly(A) region
                     /note="SV40 late poly(A) region"
     polyA_signal    3402..3523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3615..3641
                     /regulatory_class="terminator"
                     /note="E.coli"
     misc_feature    complement(3747..3949)
                     /label=human TOPO1 poly(A) region
                     /note="human TOPO1 poly(A) region"
     CDS             complement(3953..4987)
                     /codon_start=1
                     /product="hygromycin-B 4-O-kinase"
                     /label=hygromycin-B 4-O-kinase
                     /note="hygromycin-R"
                     /protein_id="BAO48053.1"
                     /translation="MDRSGKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVG
                     GRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQ
                     DLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
                     HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSE
                     AMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSL
                     VDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRP
                     RAKE"
     CDS             complement(3953..4972)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    4913
                     /label=del-Esp3I
                     /note="del-Esp3I"
     misc_feature    complement(4999..6541)
                     /label=Intron
                     /note="Intron"
     regulatory      complement(5037..5104)
                     /regulatory_class="promoter"
                     /note="E.coli EM2KC 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)
                     /regulatory_class="promoter"
                     /gene="hTOP1"
                     /note="del-ATG; human TOP1"
     gene            complement(6542..7523)
                     /gene="hTOP1"
                     /label=hTOP1
     misc_feature    6624
                     /label=del-Esp3I
                     /note="del-Esp3I"