Difference between revisions of "Part:BBa J70310:Design"

 
(→‎Source)
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 Ggagtaaattctcctttttttgg,BG7AU-F
+
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===

Revision as of 22:52, 21 July 2009

Mesoplasma florum pBG7AU plasmid origin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 99
    Illegal NheI site found at 120
    Illegal NheI site found at 141
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 779
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 747
  • 1000
    COMPATIBLE 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

References