LOCUS       Exported               10230 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pINT1, complete sequence.
ACCESSION   AF289993
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10230)
  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 10230)
  AUTHORS   Ouwerkerk PB.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUL-2000) Molecular Cell Biology, Institute of 
            Molecular Plant Sciences, Wassenaarseweg 64, Leiden 2333 AL, 
            Netherlands
REFERENCE   3  (bases 1 to 10230)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10230
                     /organism="Cloning vector pINT1"
                     /focus
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="genomic DNA"
                     /note="pINT1 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 or pHIS3NX deposited in GenBank 
                     Accession Numbers AF272029 and AF272030 respectively; pINT1
                     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 is a high copy plasmid 
                     derived from pUC29"
                     /db_xref="taxon:134364"
     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          4784..7821
                     /organism="Saccharomyces cerevisiae"
                     /chromosome="VII"
                     /mol_type="genomic DNA"
                     /label=PDC6 locus flanking sequence
                     /note="PDC6 locus flanking sequence"
                     /db_xref="taxon:4932"
     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..1440
                     /note="multiple cloning site; unique sites are NotI, XbaI, 
                     BclI"
     primer_bind     1413..1429
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(1634..1912)
                     /regulatory_class="terminator"
                     /note="yeast CYC1 gene."
     terminator      1640..1887
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(2437..3252)
                     /codon_start=1
                     /gene="apt1"
                     /product="aminoglycoside phosphotransferase"
                     /label=apt1
                     /note="APT1; similar to Escherichia coli aminoglycoside 
                     phosphotransferase: SwissProt Accession Number P00551"
                     /protein_id="AAF98176.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2437..3252)
                     /gene="apt1"
                     /label=apt1
                     /note="derived from bacterial transposon Tn903 apt1 gene"
     regulatory      complement(3268..4754)
                     /regulatory_class="promoter"
                     /note="yeast PGK1 gene"
     primer_bind     complement(7830..7846)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8324..8428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8429..9289
                     /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      9460..10048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"