pfXblue (ATCC® 77333)

Applications: expression vectorvector permitting construction of fusion proteinsvector permitting visual detection of recombinantsvector useful for cloning PCR products  /  Depositors: ME Brandt

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Designations pfXblue
Depositors ME Brandt
Biosafety Level 1
Vector Information
Size (kb): 3.6270000934600830
Vector: pfXblue (plasmid)
Promoters: Promoter lambda PL
Construction: pMb3, M13mp18, factor Xa
Marker(s):ampR
Construct size (kb): 3.627000093460083
Features: insert detection: lacZ'
marker(s): ampR
promoter: lambda PL
replicon: pMB1
Applications
expression vector
vector permitting construction of fusion proteins
vector permitting visual detection of recombinants
vector useful for cloning PCR products
Comments
Restriction digests of the clone give the following sizes (kb): BglI--1.8 (doublet); EcoRI--3.6; PstI--2.1, 1.55.
Restriction digests of the plasmid give the following sizes (kb): BamHI--3.574, 0.053; BglI--1.843, 1.784; PstI--2.072, 1.555; PvuI--2.063, 1.564; PvuII--2.972, 0.655; SacI--3.257, 0.37.
Inserts can be cloned using the 5' StuI site and one other MCS restriction site matching the 3' end of the insert. The vector supplies the AUG initiator codon.
The insert to be cloned should have a phosphorylated 5' blunt end with the first base corresponding to the first base of the first codon.
Expression vector permitting the production of a fusion protein that can be trimmed by cleavage with the coagulation factor Xa protease. A modified lacZ' allows blue/white screening of colonies.
Especially useful for the expression of PCR products.
The order of the major features of the plasmid is: pMB1 ori - ampR - lambdaPL promoter - lambda cII - factor Xa recognition sequence - StuI/MCS/HindIII/modified lacZ'.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Brandt ME, et al. A vector for directional cloning and expression of polymerase chain reaction products in Escherichia coli. Gene 97: 113-117, 1991. PubMed: 1995421

Mark E Brandt, personal communication

Shipped freeze-dried
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