yeast 2 micron A form

Vector IG Sequence Link :
General : plasmid ds-DNA 6318 BP
Functions : (cloning)
Selection : ()
Copy Number : high
Hosts : (S.cerevisiae)
Suppliers : ()
Misc.Comments : Yeast 2 micron plasmid contains two 599 bp inverted repeats separated by a large unique (UL) and a small unique (US) region. During recombination the UL and US regions invert producing two sequence forms that differ in the orientation of one unique region relative to the other. The A form is presented below. FLP is the only 2-micron circle-encoded protein needed for specific site recombination between the IRs of 2-micron circle. The minimal si ze of the recombination site required for efficient FLP recombinase-catalyzed recombination in vitro is no more than 28 bp, which includes parts of two 13 bp inverted repeats (positions 690-702 and 711-723) and all of an 8 bp spacer (703-710) [5]. The FLP recombinase cleaves the DNA at the boundaries of the spacer and becomes covalently linked to the spacer DNA [5],[9]. The efficiency of the recombination is reduced if the spacer in a recombinant site is increased or decreased by 1 bp, while the spacer in the second site is unaltered [5]. Recombination between two sites with identical 1-base pair additions or deletions is relatively unaffected, suggesting that pairing of sequences in the spacer regions is important in FLP-promoted recombinat ion events [5]. The sequence asymmetry utilized by the recombinase to determine the orientation of the site is located uniquely within the spacer region. Another 13 bp direct repeat, is found at positions 676-688 [5]. FLP-mediated recombination invo lving two FLP sites that are inverted with respect to each other results in inversion of the DNA sequences between the sites [4]. If the participating recombination sites are in direct orientation, FLP promotes only the excision of the intervening DN A sequences [4]. The Rep 1 and Rep proteins are involved plasmid partitioning and protein stability. A start codon in phase with the Rep1 coding region is located at positions 1966-1964. Two CAP sites for Rep1 mRNA are located beyond the 'atg' codon (position 2008) at positions 2004 and 2005. Complete source information: Yeast (S.cerevisiae, strain A364A D5) DNA, clones pJDB71 [1], p82-6B [2], pCV20 [3], pMMD2 [4], pGP20 [5], pJFS166 [10]. NCBI gi: 172190 & 4182
Parents : (2 micron)
Siblings : ()
Descendents : (YEp)(YIp)(2micron Circle-pBR322-LEU2)


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