|
P2300 pQuiet-3 (20mg, $)
This vector can silence the
expression of 3 genes simultaneously.?Sequences from the target genes are
used to design short cDNAs that are ligated into the insertion sites.?The
cDNAs are constructed by annealing compementary oligonucleotides to one
another.?Each oligonucleotide is designed with a 4-base overhang at its
5'-end.?The overhangs complement vector sequences that remain after
digestion with a pair of restriction enzymes that is unique for each insertion
site.?The other features of this plasmid facilitate rapid conversion to
an adenovirus using the SpeAd procedure.?This vector may be especially
useful when it is necessary to silence several genes at once to block redundant
signaling pathways.
|
Position
|
Feature
|
|
2-27
|
I-Ceu I site
|
|
30-404
|
Adenovirus nt 1-375 (L.ITR, enhancer, packaging signal)
|
|
534-539
|
Spe I site
|
|
547-552
|
Mlu I site
|
|
998-1003
|
Bgl II site
|
|
1016-1023
|
Not I site
|
|
1573-1578
|
BsrG I site
|
|
1585-1592
|
Asc I site
|
|
1594-1599
|
Cla I site
|
|
2503
|
Replication origin
|
|
3664--4075
|
Cos site
|
|
5342-4098
|
Tetracycline resistance (Tetr )
|
|
|