Difference between revisions of "Part:BBa J70592:Design"

(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
−
Designed with synthetic oligos:
+
There was a NheI site present at bp 7, so a silent mutation was introduced at that location to change a T to a G.
  
−
  5' AATTC GCGGCGC T ACTAGT TTGACGGCTAGCTCAGTCCTAGGTACAGTGCTAGC GCTAGC A GCGGCCG CTGCA 3', Prom-F
+
Designed with synthetic oligos:
−
+
  5' AATTC GCGGCCGC T ACTAGT TTGACGGCGAGCTCAGTCCTAGGTACAGTGCTAGC GCTAGC A GCGGCCG CTGCA 3', Prom-F
−
  5' GCGGCCGCTGCTAGC GCTAGCACTGTACCTAGGACTGAGCTAGCCGTCAA ACTAGTAGCGCCGCG 3', Prom-R
+
  5' GCGGCCGCTGCTAGC GCTAGCACTGTACCTAGGACTGAGCTCGCCGTCAA ACTAGTAGCGGCCGCG 3', Prom-R
  
 
Note that both primers were ordered phosphorylated. An alternative is to phosphorylate the primers yourself with a kinase.
 
Note that both primers were ordered phosphorylated. An alternative is to phosphorylate the primers yourself with a kinase.

Revision as of 13:33, 3 August 2010

RFC12 Constitutive Promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

There was a NheI site present at bp 7, so a silent mutation was introduced at that location to change a T to a G.

Designed with synthetic oligos: 

5' AATTC GCGGCCGC T ACTAGT TTGACGGCGAGCTCAGTCCTAGGTACAGTGCTAGC GCTAGC A GCGGCCG CTGCA 3', Prom-F
5' GCGGCCGCTGCTAGC GCTAGCACTGTACCTAGGACTGAGCTCGCCGTCAA ACTAGTAGCGGCCGCG 3', Prom-R

Note that both primers were ordered phosphorylated. An alternative is to phosphorylate the primers yourself with a kinase.

Source

BBa_J23100

References