LOCUS       Exported               11651 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pINT1-HIS3NB, complete sequence.
ACCESSION   AY061966
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11651)
  AUTHORS   Meijer AH, Ouwerkerk PB, Hoge JH.
  TITLE     Vectors for transcription factor cloning and target site 
            identification by means of genetic selection in yeast
  JOURNAL   Yeast 14 (15), 1407-1415 (1998)
  PUBMED    9848232
REFERENCE   2  (bases 1 to 11651)
  AUTHORS   Ouwerkerk PBF., Meijer AH.
  TITLE     Yeast one-hybrid screening for DNA-protein interactions
  JOURNAL   (in) Ausubel,F.M., Brent,R., Kingston,R.E., Moore,D.D., 
            Seidman,J.G., Smith,J.A. and Struhl,K. (Eds.); CURRENT PROTOCOLS IN 
            MOLECULAR BIOLOGY: Supplement 55, Unit 12, 12.12.1-12.12.22; John 
            Wiley & Sons, Inc., 605 Third Avenue, New York, NY, USA (2001)
REFERENCE   3  (bases 1 to 11651)
  AUTHORS   Ouwerkerk PBF., Meijer AH.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-NOV-2001) Molecular Cell Biology, Institute of 
            Molecular Plant Sciences, Wassenaarseweg 64, Leiden 2333 AL, 
            Netherlands
REFERENCE   4  (bases 1 to 11651)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11651
                     /organism="Cloning vector pINT1-HIS3NB"
                     /focus
                     /mol_type="genomic DNA"
                     /note="pINT1-HIS3NB is one of the components of an 
                     integrative vector system for one-hybrid screenings in 
                     yeast and can be used in combination with the HIS3 reporter
                     gene cassettes from plasmids pHIS3NB, pHIS3nX or pINT1, 
                     deposited in GenBank Accession Numbers AF272029 and 
                     AF272030 and AF289993; pINT1-HIS3NB is integrated by double
                     cross-over in the non-essential PDC6 locus in chromosome 
                     VII in Saccharomyces cerevisiae; transformants are selected
                     based on G418 resistance conferred by APT1 marker; 
                     pINT1-HIS3NB is a high copy plasmid derived from pUC29"
                     /db_xref="taxon:179478"
     source          275..1394
                     /organism="Saccharomyces cerevisiae"
                     /chromosome="VII"
                     /mol_type="genomic DNA"
                     /label=PDC6 locus flanking sequence
                     /note="PDC6 locus flanking sequence"
                     /db_xref="taxon:4932"
     source          6205..9242
                     /organism="Saccharomyces cerevisiae"
                     /chromosome="VII"
                     /mol_type="genomic DNA"
                     /label=PDC6 locus flanking sequence
                     /note="PDC6 locus flanking sequence"
                     /db_xref="taxon:4932"
     misc_feature    join(1..274,9243..11651)
                     /label=pUC29 vector sequence
                     /note="pUC29 vector sequence"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1405..1452
                     /note="multiple cloning site; unique sites for NotI, XbaI 
                     and SpeI"
     primer_bind     1413..1429
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            1552..2851
                     /gene="HIS3"
                     /label=HIS3
     CDS             1552..2211
                     /codon_start=1
                     /gene="HIS3"
                     /product="imidazole glycerol-phosphate dehydratase"
                     /label=HIS3
                     /note="derived from Saccharomyces cerevisiae"
                     /protein_id="AAL38968.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     regulatory      2212..2851
                     /regulatory_class="terminator"
                     /gene="HIS3"
     regulatory      complement(3055..3333)
                     /regulatory_class="terminator"
                     /note="Saccharomyces cerevisiae CYC1 terminator"
     terminator      3061..3308
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(3858..4673)
                     /codon_start=1
                     /gene="apt1"
                     /product="aminoglycoside phosphotransferase"
                     /label=apt1
                     /note="APT1; similar to SwissProt Accession Number P00551; 
                     derived from bacterial transposon Tn903 apt1 gene"
                     /protein_id="AAL38969.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(3858..4673)
                     /gene="apt1"
                     /label=apt1
     regulatory      complement(4689..6175)
                     /regulatory_class="promoter"
                     /note="Saccharomyces cerevisiae PGK1 promoter"
     primer_bind     complement(9251..9267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9745..9849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9850..10710
                     /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      10881..11469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"