pNtrp vector (V004258)

MSDS

Basic Vector Information

Vector Name:: pNtrp

Antibiotic Resistance:: Ampicillin

Length:: 8001 bp

Type:: Yeast two-hybrid vector

Replication origin:: ori

Host:: Yeast

Source/Author:: Banno H.

Promoter:: ADH1(long)

pNtrp vector Vector Map

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pNtrp vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_33612        8001 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pNtrp DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  AUTHORS   Banno H.
  TITLE     Identification of proteins that interact with a plant nuclear 
            protein using the yeast split-Trp sensor
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8001)
  AUTHORS   Banno H.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-APR-2014) Contact:Hiroharu Banno Chubu University, 
            Bioscience and Biotechnology; 1200 Matsumoto-cho, Kasugai, Aichi 
            487-8501, Japan Phone :81-568-52-6242
REFERENCE   3  (bases 1 to 8001)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8001)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-APR-2014) Contact:Hiroharu Banno Chubu University, Bioscience 
            and Biotechnology"; volume: " 1200 Matsumoto-cho, Kasugai, Aichi 
            487-8501, Japan Phone "; pages: "81-568-52-624"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..1165)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1447..1551
                     /label=AmpR promoter
     CDS             1552..2409
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2583..3171
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3479..3907
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     protein_bind    3969..3990
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4005..4035
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4043..4059
                     /label=lac operator
                     /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     4067..4083
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4125..4529
                     /label=LEU2 promoter
     CDS             4530..5621
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        6496..7200
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_feature    7209..7340
                     /label=Ntrp
                     /note="Ntrp"
     CDS             7341..7367
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_feature    7371..7423
                     /label=multiple cloning site(MCS)
                     /note="multiple cloning site(MCS)"
     primer_bind     complement(7447..7463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      7670..7857
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"

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