pRL838
(Plasmid #70680)

• Purpose: (Empty Backbone) This vector was used to generate a library of ca. 17-kb sequences of clones as part of a genomic sequence strategy and were then used to complement mutations.
• Depositing Lab: Peter Wolk
• Publication: Kaneko et al DNA Res. 2001 Oct 31;8(5):205-13; 227-53.

Backbone

• Vector backbone: modification of pBAC108L
• Backbone size (bp): 8850
• Vector type: Cre/Lox
• Selectable markers: Erythromycin for selection in cyanobacteria

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Cloning Information

• Cloning method: Restriction Enzyme

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Shizuya, H., Birren, B., Kim, U. J. et al. 1992, Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector., Proc. Natl. Acad. Sci. USA, 89, 8794–8797

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

To facilitate the construction of low-copy-number BAC vector pRL838 (GenBank AF403425), a derivative of pBAC108L and pUC19 were fused at their unique BspLU11I (isoschizomer of PciI) sites, permitting modification of the BAC vector as a high-copy-number plasmid. The BAC vector was later freed from pUC19 by use of BspLU11I. To take further advantage of the features bracketing the cloning region of pBAC108L, the Sal I and Sst I sites were relocated to the polylinker, and other unique sites added. With the exception of BanII (GRGCY/C), but including its SacI subset (GAGCT/C) and BspLU11I (not in the polylinker), the ends of the large restriction fragments--from 0.1 to more than 0.5 Mb--resulting from digestion by 16 restriction enzymes (Fig. 2 of Bancroft et al. J Bacteriol 171: 5940-5948, 1989) could be accommodated by the restriction sites of the polylinker.

Additional references:
Wolk CP, Fan Q, Zhou R, Huang G, Lechno-Yossef S, Kuritz T, Wojciuch E (2007) Paired cloning vectors for complementation of mutations in the cyanobacterium Anabaena sp. strain PCC 7120. Arch Microbiol 188: 551-563
Huang G, Fan Q, Lechno-Yossef S, Wojciuch E, Wolk CP, Kaneko T, Tabata S (2005)
Clustered genes required for the synthesis of heterocyst envelope polysaccharide in
Anabaena sp. strain PCC 7120. J Bacteriol 187: 1114-1123
Fan Q, Huang G, Lechno-Yossef S, Wolk CP, Kaneko T, Tabata S (2005) Clustered genes
required for synthesis and deposition of envelope glycolipids in Anabaena sp. strain PCC
7120. Mol Microbiol 58: 227-243