LOCUS       Exported                8570 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Signal sequence gene trap vector pSA-TMbgyg, complete sequence.
ACCESSION   AB183486
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8570)
  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 8570)
  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 8570)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8570
                     /organism="Signal sequence gene trap vector pSA-TMbgyg"
                     /mol_type="other DNA"
                     /db_xref="taxon:284807"
     misc_feature    17..120
                     /label=splicing acceptor
                     /note="splicing acceptor"
     CDS             96..4908
                     /codon_start=1
                     /product="fusion protein"
                     /label=fusion protein
                     /protein_id="BAD83863.1"
                     /translation="VSTPRSNCSRRYRGPHGRVEMEGREGSFFPVLDHLLPKEPKGVCA
                     EVYQQPQVPAVRNAPTHPSDTPGLPSVCWFWQPDPDSGQRDTVSGSEPGGDESDSARQQ
                     HFDL*GDGTYLTQDETDLEAGESGGQGLQAGESDSSAGP*SRGVAMSTE*R*RGQDGLQ
                     DPGFIQRVEPDNVPGCRAGERLQLSGFHGALHPILCQVPAPTAPGSTDVSDQEASQ*EE
                     DLPVLPPDAEKPQSHGSRRFTTS*LGKPWRYPT*SPCSTSPFRQLA**RRGPHRSPFPT
                     VAQPEWRMALCLVSGTRSGAGKLAGVRSS*GRYCRRPLKLADARLRCAHLHQRNLSHYG
                     QSAVCSHGESDGLLLAHI*C**KLATGRPDANYF*WR*LGVSSVVQRALGRLRPGQSFA
                     V*I*PERIFTRRRKPPRGDGAALE*RQLSGRSGYVADERHFP*RLVAA*TDYTNQRFPC
                     CHSL***FQPRCTGG*SSDVRRVA*LPTGNSFFMAG*NAGRQRHRAFRR*NYR*AWWLC
                     RSRHTTSERRKPETVERRNPESLSCGG*TAHRRRHAD*SRSLRCRFPRGAD*KWSAAAE
                     RQAVADSRR*PSRASSSAWSGHG*ADDGAGYPADEAEQL*RRALFALSEPSAVVHAVRP
                     LRPVCGG*SQY*NPRHGANESSDR*SALATGDERTRNANGAARS*SPECDHLVAGE*IR
                     PRR*SRRAVSLDQICRSFPPGAV*RRRSRHHGHRYYLPDVRARG*RPALPGCAEMVHQK
                     MAFATWRDAPADPLRIRPRDG*QSWRFR*ILAGVSSVSPFTGRLRLGLGGSVAD*I**K
                     RQPVVGLRR*FWRYAERSPVLYERSGLCRPHAASSADGSKTPAAVFPVPFIRANHRSDQ
                     RIPVPS*R*RAPALDGGAGW*AAGKR*SASGCRSTR*TVD*TA*TTAAGERRATLAHST
                     RSATERDRMVRSRAHQRLAAVASGGKPQCDAPRRVPRHPASDHQRNGFLHRAG**ALAI
                     *PPVRLSFTDVDWR*KTTADAAARSVHPCTAG*RHWRK*SDPH*P*RLGRTLEGGGPLP
                     GRSSVVAVHGRYTC*CGADYDRSRVAASGENLIYQPENLPD*W*WSNGDYR*C*SGERY
                     TASGADWPELPAGAGSRAGKLARIRAARKLSRPPYCRLF*PLGSAIVRHVYPVRLPERK
                     RSALRDARIELWPTPVARRLPVQHQPLQSTATDGNQPSPSAARGRRHMAEYRRFPYGDW
                     WRRLLEPVSIGGIPAERRSLPLPVGLVSGDPRHHEKA*THRDVCREVSDRKVRQRLRPD
                     AALGGRRISCFQLRCRRAWICPAGK*LRRWFLQRSLCLSALCIGRAPDSGSA*HWGIQR
                     EPDLLHLPPCTGCHVARPA*NRTARCSAAGRGGHGCDRCGRS*PDERVRPIRTARNRSI
                     HYMA*FHMRDC*SPCVSLANCDGRHRQCVRRAGSR*ADALGRGLPRSPAPRARGFRLQQ
                     CPDGQWPHNSGH*LERGDVRGFPIRGRQHLLLEAVVGLYGAADALLRAEASGACRIAAA
                     PGVYAPHWS*PTLSELG*RQFR*CSLGAGSMRRNRPIRSRDCRAYTNRPQKRGRLDRWL
                     CRSTRR*WKPTPQHSSGSGDGGG*L"
     misc_feature    144..804
                     /gene="TM(CD4)"
                     /product="transmembrane domain of rat CD4"
     gene            144..804
                     /gene="TM(CD4)"
                     /label=TM(CD4)
     misc_feature    808..3854
                     /gene="LacZ"
                     /product="beta-galactosidase"
     gene            808..3854
                     /gene="LacZ"
                     /label=LacZ
     CDS             814..3855
                     /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    3871..4908
                     /gene="hyg"
                     /product="phosphotransferase"
     gene            3871..4908
                     /gene="hyg"
                     /label=hyg
     CDS             3871..4908
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     regulatory      4909..5188
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    4950..4998
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      5189..5408
                     /regulatory_class="terminator"
     promoter        complement(5439..5457)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5475..5491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5499..5515
                     /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(5523..5553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5568..5589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5877..6465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        complement(7497..7601)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7935..8390
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8531..8547
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8554..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"