LOCUS       Exported                5890 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pInSRT-GFPhFDC.SP DNA, complete sequence.
ACCESSION   AB375115
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5890)
  AUTHORS   Shinomura T, Nakamura S, Ito K, Shirasawa S, Hook M, Kimura JH.
  TITLE     Adsorption of Follicular Dendritic Cell-secreted Protein (FDC-SP) 
            onto Mineral Deposits: APPLICATION OF A NEW STABLE GENE EXPRESSION 
            SYSTEM
  JOURNAL   J. Biol. Chem. 283 (48), 33658-33664 (2008)
  PUBMED    18806264
REFERENCE   2  (bases 1 to 5890)
  AUTHORS   Shinomura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JAN-2008) Contact:Tamayuki Shinomura Tokyo Medical and
            Dental University, Graduate School, Tissue Regeneration; 1-5-45 
            Yushima, Bunkyo-ku, Tokyo 113-8549, Japan
REFERENCE   3  (bases 1 to 5890)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5890
                     /organism="Cloning vector pInSRT-GFPhFDC.SP"
                     /mol_type="other DNA"
                     /db_xref="taxon:495548"
     promoter        14..32
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    50..97
                     /label=FRT (FLP recombination target)
                     /note="FRT (FLP recombination target)"
     protein_bind    50..97
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             191..1216
                     /codon_start=1
                     /gene="GFPhFDC.SP"
                     /product="GFP-FDC.SP fusion protein"
                     /label=GFPhFDC.SP
                     /note="green fluorescence protein (GFP)-follicular 
                     dendritic cell secreted peptide (FDC.SP) fusion protein"
                     /protein_id="BAG54842.1"
                     /translation="MFINIKSILWMCSTLIVTHALHKVKVGKSPPVATMVSKGEELFTG
                     VVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQ
                     CFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGI
                     DFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTP
                     IGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYRSPVSQDQE
                     REKRSISDSDELASGFFVFPYPYPFRPLPPIPFPRFPWFRRNFPIPIPESAPTTPLPSE
                     K"
     gene            191..1216
                     /gene="GFPhFDC.SP"
                     /label=GFPhFDC.SP
     regulatory      287..296
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             293..1006
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     misc_feature    1258..1844
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1845..2444
                     /codon_start=1
                     /gene="puro"
                     /product="puromycin phosphotransferase"
                     /label=puro
                     /protein_id="BAG54843.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            1845..2444
                     /gene="puro"
                     /label=puro
     polyA_signal    2531..2755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      2621..2626
                     /regulatory_class="polyA_signal_sequence"
     promoter        complement(2789..2807)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2814..2830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2971..3426)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3760..3864
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4896..5484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5772..5793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5808..5838
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5846..5862
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     5870..5886
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"