P-296 cpDNA XhoI 8 (9.5 kb) er-u-1a
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Boynton-Gillham laboratory, Duke University, 1990
Chloroplast DNA XhoI 8 (9.5 kb) in BSKS+; psbA through 3′ exon of 23S; ca. 1.5 kb Kpn fragment was deleted between psbA and 5S;; contains er-u-1a erythromycin resistance mutation in 23S, from CC-64
host strain: JM109
amp resistant
More information
Strain: CC-64 er-u-1a mt+
Insert: XhoI 8 ΔKpnI (9.5 kb)
Genes present: psbA, 5S rRNA, 23S rRNA (3′ exon)
Vector: BSKS+
Bacterial host strain: JM109
Selectable marker: amp-r, white on X-gal
Origin: Amnon Lers in Boynton/Gillham lab, 1990
The insert is a subclone of Xho 8 (P-259B) in which three KpnI fragments including the BamHI site (approx. 1.5 kb) located between the psbA and 5S genes were deleted and the remaining two Kpn ends were religated, leaving only one Kpn site.
The er-u-1a mutation is a C->T change at bp 2622 in C. reinhardtii 23S rRNA gene (Lemieux numbering, equivalent to E. coli 2611), which confers erythromycin resistance in Chlamydomonas.