Sequence of YEp13

Suzuki Chise csuzuki at nfri.affrc.go.jp
Sat Nov 11 02:44:45 EST 1995


[summary of YEp13 sequence]
There are some kinds of YEp13 sequence data.
1. From Genbank (U03498)
	This sequence (10667 bp) has no Sac I site.
2. http://biology.queensu.ca/~miseners/vector_descrip/YEP13.html
	This sequence (10639 bp) has two groups of "nnn" for the un-
	sequenced region (see below).
3. Method Enz. vol.194, p.207 (restriction map)
	This map (10786 bp) has a Sac I site at 8392.

We do not have a correct map of YEp13, so far. 
 
Following is the mail from Dr. David Stillman, which is very useful 
for YEp13 users. I would like to thank Francis and David for their 
help.
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Concerning YEp13: 

Several years ago I made a computer version of YEp13 much like the one at
http://biology.queensu.ca/~miseners/vector_descrip/YEP13.html. YEp13 is
pBR322, with a 2 micron fragment inserted, and then a 4.1 kb PstI fragment
with LEU2 inserted. Unfortunately only 2.2 kb of this 4.1 kb PstI fragment
was sequenced, and so my vector had two groups of "nnn" for the un-sequenced
region. 

Two years ago I decided to make the complete YEp13 vector sequence, using the
sequence of the LEU2 region on Chromosome III. Unfortunately, the chromosome
III sequence does not contain two appropriately positioned PstI sites. It
does contain one PstI site at the appropriate postion (nt 93773). I assumed
that there was restriction site polymorphism between the yeast strain that
was sequenced and the strain from which the 4.1 kb LEU2 was isolated. So I
searched in the region 4.1 kb upstream of nt 93773 for a 5 out of 6 match to
the PstI site, and found one at 89701. By converting the sequence CTACAG at
89701 to CTGCAG (PstI site), I could use this 4072 fragment to complete the
vector. 
The restiction maps of YEp13, made long before wide scale seqeuncing, also
show BglII sites in each of the unsequenced regions (the 585 bp PstI - XhoI
and the 1265 SalI - PstI segments). Importantly, there are BglII sites in
each of these regions in my computer version.

Where is the SacI site? Good question. Another restriction site polymorphism?

I hope this information is useful. 

David
===========================
David Stillman
Division of Molecular Biology and Genetics
Department of Oncological Sciences
University of Utah Heath Sciences Center 
Salt Lake City, UT  84132
stillman at bioscience.utah.edu
phone: 801-581-5429
fax: 801-581-3607

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Chise Suzuki, Ph.D
National Food Research Institute
email:csuzuki at nfri.affrc.go.jp
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