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Size (kb): 5.5000000000000000
Vector: pJFCAT1 (phagemid)
Construction: pBLCAT3, pUCf1, SV40 poly(A)
Construct size (kb): 5.5
Features: insert detection: CAT
replicon: pMB1, f1
vector containing primer sites useful for sequencing
vector permitting production of single-stranded DNA
Restriction digests of the clone give the following sizes (kb): HindIII--5.0, 0.75; EcoRI/HindIII--2.7, 2.0, 0.75, 0.3; XhoI--5.8.
A promoter/enhancer-cloning shuttle vector using chloramphenicol acetyltransferase (CAT) as the reporter gene.
The KpnI and SstI sites can be used for cloning enhancer elements.
To avoid potential problems using primer 2, try the following primer sequence instead (assumes promoter inserted at or downstream of the SphI site): 5' TTATCATGTCTGGATCCAAG 3'
Promoter elements cloned into the polylinker may be sequenced using the following oligonucleotide primers. primer 1: 5'-TCCTTAGCTCCTGAAAATCT- 3'; primer 2: 5'-AAACTCATCAATGTATCTTA- 3'.
Contains a trimer cassette of the SV40 major late polyadenylation signal to block background readthrough expression of the reporter gene.
The order of the major features in this phagemid is : HindIII - SV40 polyadenylation trimer - primer 2 site - BamHI/MCS/XhoI - primer 1 site - CAT gene - SV40 splice and polyadenylation signal - f1 ori - KpnI - SstI - ampR/pUC18.
Fridovich-Keil JL, et al. Improved expression vectors for eukaryotic promoter/enhancer studies. BioTechniques 11: 572-579, 1991. PubMed: 1725108
Judith L Fridovich-Keil, personal communication