CGSC Strain#: 7629
Strain Designation: BT340     
Source of Strain: W. Wackernagel
Other Designations: DH5alpha/pCP20
Other Designations: pCP20
Sex: F-
Episome/Plasmid: pCP20
Plasmid Markers/Mutations: [cI857](λ)(ts)
,
bla(ApR)
,
cat,
[FLP],
repA101(ts)
,
FLP ,
Cm ,
Ap Plasmid Comment: pCP20 has the yeast Flp recombinase gene, FLP, chloramphenicol and ampicillin resistant genes, and temperature sensitive replication.
Chromosomal Markers: Δ(argF-lac)169, φ80d
lacZ58(M15),
glnX44(AS)
,
λ-,
rfbC1,
gyrA96(NalR)
,
recA1,
endA1,
spoT1,
thiE1,
hsdR17Strain Comments: - 1.This is a recA- strain and grows slowly, and this is even more noticeable at 30C, of course. (It may be best to ``revive'' the culture, if shipped on sterile disks, without antibiotics and
- 2.grow next transfer on amp or amp + Cm.)
- pCP20 carries the yeast Flp recombinase gene, (FLP, aka exo), chloramphenicol and ampicillin resistant genes, and temperature sensitive replication.
- This strain should be grown on rich media with ampicillin (100 ug/ml) or chloramphenicol (25 ug/lml), or both, at 30 C.
- Δ(argF-lac)169-- extends from mmuP through orfs preceding argF, through lac to mhpD, literally Δ(mmuP-mhpD)169. (Peters et al. 2003 JB 185:2017)
- Δ(argF-lac)169-- from strain Hfr3000 U169 was initially called ΔlacU169 and described as a lacZY mutation until found to include argF and lacI.
- glnX44(AS)-- Mutation is a C to T transition at nucleotide 34 (3rd position of the anticodon)
- gyrA96(NalR)-- resistant to 200ug/ml nalidixic acid
- gyrA96(NalR)-- GAC to AAC at codon 87
- recA1-- : Missense mutation, altered isoelectric point. Sequenced: G to A for Nuc. 720 (Gly 160 Asp).
- endA1-- G to A transition mutation resulting in E208K amino acid substitution.
- spoT1-- In addition to the 6bp insertion, there is a C to T transition resulting in an H257Y substitution.
- glnX44(AS) was formerly called su+II
- glnX44(AS) was formerly called supE44
- glnX44(AS) was formerly called glnV44
- gyrA96(NalR) was formerly called
- rfbC1 was formerly called rfbD1
- thiE1 was formerly called thi-1
- = good miniprep dsDNA quality
- AS = amber(UAG) suppressor
- NalR = Nalidix.acidResist.
- ApR = ampicillin resistant
- ts = Temperature Sensitive
References: - Cherepanov, PP, W Wackernagel 1995. Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene 158(1):9-14.