LOCUS       Exported                9657 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Eukaryotic vector pSV-ssTMbgyg, complete sequence.
ACCESSION   AB183487
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9657)
  AUTHORS   Shirasawa S, Yoshimi M, Kamochi H, Muneta T, Kimura JH, Hook M, 
            Shinomura T.
  TITLE     Gene trap screening for cell surface and extracellular matrix 
            molecules produced by chondrocytes
  JOURNAL   J. Biochem. 137 (1), 79-85 (2005)
  PUBMED    15713886
REFERENCE   2  (bases 1 to 9657)
  AUTHORS   Shinomura T, Shirasawa S.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUL-2004) Tamayuki Shinomura, Tokyo Medical and Dental
            University, Tissue Regeneration, Dept. of Hard Tissue Engineering; 
            1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan 
            (E-mail:t.shinomura.trg@tmd.ac.jp, Tel:81-3-5803-5450, 
            Fax:81-3-5803-0187)
REFERENCE   3  (bases 1 to 9657)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9657
                     /organism="Eukaryotic vector pSV-ssTMbgyg"
                     /mol_type="other DNA"
                     /db_xref="taxon:284808"
     regulatory      45..384
                     /regulatory_class="promoter"
                     /standard_name="SV40 promoter"
     promoter        157..367
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        179..375
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      226..361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          401..973
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1106..5995
                     /codon_start=1
                     /product="fusion protein"
                     /label=fusion protein
                     /protein_id="BAD83864.1"
                     /translation="MSALLILALVGAAVADYKDDDDKLAAANSSIDRSDIGTAGPMGEL
                     RWKAEKAPSSQSWITFSLKNQKVSVQKSTSNPKFQLSETLPLTLQIPQVSLQFAGSGNL
                     TLTLDRGILYQEVNLVVMKVTQPDSNTLTCEVMGPTSPKMRLILKQENQEARVSRQEKV
                     IQVQAPEAGVWQCLLSEGEEVKMDSKIQVLSKGLNQTMFLAVVLGSAFSFLVFTGLCIL
                     FCVRCRHQQRQAARMSQIKRLLSEKKTCQCSHRMQKSHNLMDPVVLQRRDWENPGVTQL
                     NRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADT
                     VVVPSNWQMHGYDAPIYTNVTYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRII
                     FDGVNSAFHLWCNGRWVGYGQDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQD
                     MWRMSGIFRDVSLLHKPTTQISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSL
                     WQGETQVASGTAPFGGEIIDERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTAD
                     GTLIEAEACDVGFREVRIENGLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILL
                     MKQNNFNAVRCSHYPNHPLWYTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMS
                     ERVTRMVQRDRNHPSVIIWSLGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTAT
                     DIICPMYARVDEDQPFPAVPKWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQA
                     FRQYPRLQGGFVWDWVDQSLIKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPH
                     PALTEAKHQQQFFQFRLSGQTIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVA
                     PQGKQLIELPELPQPESAGQLWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPA
                     ASHAIPHLTTSEMDFCIELGNKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDN
                     DIGVSEATRIDPNAWVERWKAAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLF
                     ISRKTYRIDGSGQMAITVDVEVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDR
                     LTAACFDRWDLPLSDMYTPYVFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLM
                     ETSHRHLLHAEEGTWLNIDGFHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQGILAT
                     MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYVLRVNSCADGFYK
                     DRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEA
                     MDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQA
                     LDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWR
                     PWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDA
                     IVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDREMGEAN"
     gene            1106..1150
                     /gene="ss"
                     /label=ss
     sig_peptide     1106..1150
                     /gene="ss"
     CDS             1106..1150
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1151..1174
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1231..1893
                     /gene="TM(CD4)"
                     /product="transmembrane domain of rat CD4"
     gene            1231..1893
                     /gene="TM(CD4)"
                     /label=TM(CD4)
     misc_feature    1894..4941
                     /gene="LacZ"
                     /product="beta-galactosidase"
     gene            1894..4941
                     /gene="LacZ"
                     /label=LacZ
     CDS             1901..4942
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQ"
     misc_feature    4958..5995
                     /gene="hyg"
                     /product="phosphotransferase"
     gene            4958..5995
                     /gene="hyg"
                     /label=hyg
     CDS             4958..5995
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     regulatory      5996..6275
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    6037..6085
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      6276..6495
                     /regulatory_class="terminator"
     promoter        complement(6526..6544)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6562..6578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6586..6602
                     /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)."
     promoter        complement(6610..6640)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6655..6676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6964..7552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        complement(8584..8688)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9022..9477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     9618..9634
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9641..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"