pHSG396 vector (V012464)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V012464	pHSG396	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pHSG396

Antibiotic Resistance:: Chloramphenicol

Length:: 2238 bp

Type:: Cloning Vectors

Replication origin:: ori

Source/Author:: Takeshita S, Sato M, Toba M, Masahashi W, Hashimoto-Gotoh T.

Copy Number:: High copy number

5' Primer:: M13 fwd

3' Primer:: M13 rev

pHSG396 vector Map

Copy Sequence View Map

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.

pHSG396 vector Sequence

LOCUS       pHSG396.        2238 bp DNA     circular SYN 01-JAN-1980
DEFINITION  pUC-type bacterial cloning vector with a chloramphenicol resistance 
            gene. The MCS is similar but reversed in pHSG398.
ACCESSION   .
VERSION     .
KEYWORDS    pHSG396
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2238)
  AUTHORS   Takeshita S, Sato M, Toba M, Masahashi W, Hashimoto-Gotoh T.
  TITLE     High-copy-number and low-copy-number plasmid vectors for lacZ 
            alpha-complementation and chloramphenicol- or kanamycin-resistance 
            selection.
  JOURNAL   Gene 1987;61:63-74.
  PUBMED    3327753
REFERENCE   2  (bases 1 to 2238)
  AUTHORS   TaKaRa
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 2238)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1987"; volume: "61"; pages: "63-74"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..2238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..192
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             193..849
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     protein_bind    1059..1080
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1095..1125
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1133..1149
                     /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     1157..1173
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1186..1254
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     primer_bind     complement(1255..1271)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1597..2185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"