Difference between revisions of "Part:BBa J70310:Design"
(→Source) |
|||
(One intermediate revision by the same user not shown) | |||
Line 1: | Line 1: | ||
− | |||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_J70310 short</partinfo> | <partinfo>BBa_J70310 short</partinfo> | ||
Line 17: | Line 16: | ||
Primers: | Primers: | ||
− | GTTTCTTC GAATTC GCGGCCGC T TCTAGA | + | GTTTCTTC GAATTC GCGGCCGC T TCTAGA Gctcctttttttggggttagcgatag,BG7AU-F2 |
GTTTCTTC CTGCAG CGGCCGC T ACTAGT Acaaaaaaaggagaatttactcc,BG7AU-R | GTTTCTTC CTGCAG CGGCCGC T ACTAGT Acaaaaaaaggagaatttactcc,BG7AU-R | ||
+ | |||
+ | Original part design used this primer, which had primer-dimer issues with -R primer. | ||
+ | Redesign using -F2 primer, modified the part sequence at the 5' end. | ||
+ | |||
+ | GTTTCTTC GAATTC GCGGCCGC T TCTAGA Ggagtaaattctcctttttttgg,BG7AU-F | ||
===References=== | ===References=== |
Latest revision as of 22:53, 21 July 2009
Mesoplasma florum pBG7AU plasmid origin
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 99
Illegal NheI site found at 120
Illegal NheI site found at 141 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 779
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 747
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Unclear location for the cut site of the circular plasmid.
Source
PCR from recircularized pBG7AU cut at the ClaI site. Original plasmid from Djordjevic01. Primers:
GTTTCTTC GAATTC GCGGCCGC T TCTAGA Gctcctttttttggggttagcgatag,BG7AU-F2
GTTTCTTC CTGCAG CGGCCGC T ACTAGT Acaaaaaaaggagaatttactcc,BG7AU-R
Original part design used this primer, which had primer-dimer issues with -R primer.
Redesign using -F2 primer, modified the part sequence at the 5' end.
GTTTCTTC GAATTC GCGGCCGC T TCTAGA Ggagtaaattctcctttttttgg,BG7AU-F