YIplac204 vector (V010162)

Basic Vector Information

      • Vector Name:
      • YIplac204
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 3545 bp
      • Type:
      • Yeast Plasmids
      • Source/Author:
      • Gietz RD, Sugino A.
      • Copy Number:
      • High copy number
      • 5' Primer:
      • M13 fwd
      • 3' Primer:
      • M13 rev

YIplac204 vector Vector Map

YIplac2043545 bp6001200180024003000lac promoterlac operatorM13 revARS1TRP1 promoterAmpR promoterAmpRori

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 ℃.

YIplac204 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                3545 bp ds-DNA    circular SYN 29-11-2013
DEFINITION  Yeast integrating plasmid with a TRP1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    YIplac204
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3545)
  AUTHORS   Gietz RD, Sugino A.
  TITLE     New yeast-Escherichia coli shuttle vectors constructed with in vitro
            mutagenized yeast genes lacking six-base pair restriction sites.
  JOURNAL   Gene 1988;74:527-34.
  PUBMED    3073106
REFERENCE   2  (bases 1 to 3545)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     The sequence of this plasmid was reconstructed using the description
            from Gietz and Sugino.
FEATURES             Location/Qualifiers
     source          1..3545
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     promoter        142..172
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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     204..220
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             216..539
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    233..289
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC19 multiple cloning site"
     primer_bind     complement(290..306)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      696..936
                     /note="ARS1"
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(775..1449)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(1450..1551)
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     promoter        1639..1743
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1744..2604
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2775..3363
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"

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