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MP-P8 vector (V016992)

Price Information

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V016992 MP-P8 In stock, instant shipping

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

Basic Vector Information

Vector Name:
MP-P8
Antibiotic Resistance:
Chloramphenicol
Length:
6651 bp
Type:
Protein expression
Replication origin:
CloDF13 ori
Host:
E. coli
Promoter:
araBAD
Growth Strain(s):
DH5a
Growth Temperature:
37℃

MP-P8 vector Map

Copy Sequence View Map
MP-P86651 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600CloDF13 orirrnB T1 terminatoraraCaraBAD promoterDNA polymerase III subunit epsilonDNA adenine methylaseNegative modulator of initiation of replicationTranscriptional repressor MprAUGIAIDtonB terminatorCmR

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.

MP-P8 vector Sequence

LOCUS       V016992                 6651 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016992
VERSION     V016992
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6651)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(39..777)
                     /direction=LEFT
                     /label="CloDF13 ori"
                     /note="Plasmids containing the CloDF13 (CDF) origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     terminator      complement(870..913)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(930..1805)
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     promoter        1832..2116
                     /label="araBAD promoter"
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite
                     direction (Guzman et al., 1995)"
     CDS             2166..2894
                     /gene="dnaQ"
                     /label="DNA polymerase III subunit epsilon"
                     /note="DNA polymerase III subunit epsilon from Escherichia
                     coli (strain K12). Accession#: P03007"
     CDS             2926..3759
                     /gene="dam"
                     /label="DNA adenine methylase"
                     /note="DNA adenine methylase from Escherichia coli (strain
                     K12). Accession#: P0AEE8"
     CDS             3791..4333
                     /gene="seqA"
                     /label="Negative modulator of initiation of replication"
                     /note="Negative modulator of initiation of replication from
                     Escherichia coli (strain K12). Accession#: P0AFY8"
     CDS             4365..4892
                     /gene="mprA"
                     /label="Transcriptional repressor MprA"
                     /note="Transcriptional repressor MprA from Escherichia coli
                     (strain K12). Accession#: P0ACR9"
     CDS             4924..5175
                     /label="UGI"
                     /note="uracil-DNA glycosylase inhibitor from a Bacillus
                     subtilis bacteriophage (Mol et al., 1995)"
     CDS             5208..5801
                     /label="AID"
                     /note="human activation-induced cytidine deaminase
                     (Dickerson et al., 2003)"
     terminator      5831..5862
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     CDS             complement(5881..6537)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"