LOCUS       Exported                9004 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBC214, complete sequence.
ACCESSION   AF474332
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9004)
  AUTHORS   Frieman MB, McCaffery JM, Cormack BP.
  TITLE     Modular domain structure in the Candida glabrata adhesin Epa1p, a 
            beta1,6 glucan-cross-linked cell wall protein
  JOURNAL   Mol. Microbiol. 46 (2), 479-492 (2002)
  PUBMED    12406223
REFERENCE   2  (bases 1 to 9004)
  AUTHORS   Cormack BP, Frieman MB.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2002) Molecular Biology and Genetics, Johns 
            Hopkins University, 725 North Wolfe Street, Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 9004)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9004
                     /organism="Cloning vector pBC214"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:187395"
     misc_feature    68..571
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        608..712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             713..1573
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1744..2332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2656..2686
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2694..2710
                     /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     2718..2734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2755..2773
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3193..6408
                     /codon_start=1
                     /product="HA-tagged epithelial adhesion 1"
                     /label=HA-tagged epithelial adhesion 1
                     /note="derived from Candida glabrata; HA-EPA1; cell wall 
                     protein"
                     /protein_id="AAL84600.1"
                     /translation="MILNPALFLNKCVCIYTTLILLLLTNGGYATSTIFYPYDVPDYAG
                     YPYDVPDYAGSYPYDVPDYAAQCSTNDISLASKDPTTFPLGCSPDITTPKKGLSMELYS
                     YDFRKKGSYPCWDAAYLDPNYPRTGYKSHRLLAKVDGVTGNINFYYHATKGCTPQLGHL
                     PASYNYPKPLTMTNFTMLLYGYFRPKVTGFHTFTISADDLLFVNFGAGNAFDCCRRDSS
                     ADHFGNYQAYAIWGSKTAKDELTVHLDAGVYYPIRLFYNNREYDGALSFTFKTESNENT
                     VSDFSEYFFSLDDTEEGCPGLISYDSSCASVKTSKIIGIDYHTETPNENLVPITKTIYH
                     LGIPCTGTTTTPLCGSGFYDPLANKCVTINTSSTSSVTKTTSHTTSKEVSFHSSISSQK
                     TLIPKSIPSPYGPIKSQSIPTEMETSSEISSSEYAFSDVISTPSHSPYTKKHSSLNSSS
                     YTSTVIHSLTSYSISQGIFSTSLSEQNITSKSSTDKFSTATSMSNNITQSSIIISQFST
                     NNENYTTTSMHTSSDKISTETLNNSISTTTSILFSNSSTILKNNTTIISSDKDTHYHPV
                     NPTIVCSTNKTEIICASITQPSISNSNNHWSSSVLKFNSTTVRSTLPSSAGSNETSINV
                     PFSSSTESNTSTSSTSTSNSKMVRSTLPSSAGSNETSISVPFSSSTESNTSTSSTSTSN
                     SKMVRSTLPSSAGSNETSINVPFSSSTESNTSTSSTSTSNSKTVRSTPFSSAGIIMTSL
                     SQRNNKSASSYASSNSKCYNTADSCRKVHSTPSYLLTSSYTSEGVDYDCSLVSTKLKIN
                     DTNCVNNKHTTKSCLKTSVTTTIPTLEIKTARKSSSNTIGLHSYPTSSPNKSISSAPII
                     GYISSFKTIKTASPSYQTSDLTTITTITSLNNPGSTAVENTHESNDKSRKTSSNDISSK
                     HSVIKETKDAVESSNKSHQTNTLKCSSIIIASSSHNSYESLGGTTLTLTLSKVYSPQNN
                     DTLPFLEISEVNPSRTVLPESSKMMQYLTSTEERNKTARNTIATNIVSISTFHFEGEGN
                     AIRMGYTQLLLMLIGIIVMNIGT"
     CDS             3298..3387
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(6719..6735)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6742..6991
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(7010..7028)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7038..7054)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7199..7654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(7785..8588)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(8589..8804)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter