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pPink-HC vector (V010237) Gene synthesis in pPink-HC backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V010237 pPink-HC In stock, instant shipping

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pPink-HC
Antibiotic Resistance:
Ampicillin
Length:
7667 bp
Type:
Yeast Plasmids
Replication origin:
ori
Source/Author:
Invitrogen (Life Technologies)
Copy Number:
High copy number
Promoter:
AOX1
Growth Strain(s):
DH10B

pPink-HC vector Map

Copy Sequence View Map
pPink-HC7667 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500AOX1 promoterCYC1 terminatorPpADE2 HC promoterPpADE2PpTRP2 promoterPpTRP2M13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterM13 fwd

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pPink-HC vector Sequence

LOCUS       Exported                7667 bp DNA     circular SYN 21-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pPink-HC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7667)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7667)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     terminator      1017..1266
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        1286..1298
                     /gene="Pichia pastoris ADE2"
                     /label=Pichia pastoris ADE2 promoter
                     /note="PpADE2 HC promoter"
                     /note="weak 13-bp promoter fragment that requires high-copy
                     integration for efficient expression"
     CDS             1299..2987
                     /label=PpADE2
                     /note="phosphoribosylaminoimidazole carboxylase from Pichia
                     pastoris"
     promoter        3217..3366
                     /label=PpTRP2 promoter
                     /note="promoter for Pichia pastoris TRP2"
     CDS             3367..4980
                     /label=PpTRP2
                     /note="anthranilate synthase from Pichia pastoris"
     primer_bind     complement(5045..5061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5069..5085)
                     /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)."
     promoter        complement(5093..5123)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5138..5159)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5447..6035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6209..7066)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7067..7171)
                     /label=AmpR promoter
     primer_bind     7645..7661
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"