Difference between revisions of "Part:BBa K553000:Design"

(→‎Design Notes)
(→‎Design Notes)
Line 7: Line 7:
 
===Design Notes===
 
===Design Notes===
  
−
the part was generated by per with these primers:
+
the part was generated by using the following primers:
  
−
'''TraR rev '''
 
−
 
−
5' GTTTCTTCCTGCAGCGGCCGCTACTAGTATCAGATGAGTTTCCGCCGGATG  3'
 
  
 
'''TraR fw '''
 
'''TraR fw '''
Line 17: Line 14:
 
5' GTTTCTTCGAATTCGCGGCCGCTTCTAGATCGACGACTGGCTGGACAAGC    3'
 
5' GTTTCTTCGAATTCGCGGCCGCTTCTAGATCGACGACTGGCTGGACAAGC    3'
  
−
with the present protocol:
+
'''TraR rev '''
 +
 
 +
5' GTTTCTTCCTGCAGCGGCCGCTACTAGTATCAGATGAGTTTCCGCCGGATG  3'
 +
 
 +
 
 +
pcr protocol:
  
 
95° 5' | 10x (93° 30" | 56° 30" | 72° 40") | 23x ( (93° 30" | 65° 30" | 72° 40") | 72° 7' | 4° ∞
 
95° 5' | 10x (93° 30" | 56° 30" | 72° 40") | 23x ( (93° 30" | 65° 30" | 72° 40") | 72° 7' | 4° ∞

Revision as of 13:27, 19 September 2011

TraR trans-activator


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

the part was generated by using the following primers:


TraR fw

5' GTTTCTTCGAATTCGCGGCCGCTTCTAGATCGACGACTGGCTGGACAAGC 3'

TraR rev

5' GTTTCTTCCTGCAGCGGCCGCTACTAGTATCAGATGAGTTTCCGCCGGATG 3'


pcr protocol:

95° 5' | 10x (93° 30" | 56° 30" | 72° 40") | 23x ( (93° 30" | 65° 30" | 72° 40") | 72° 7' | 4° ∞


The part contains also the BBa_B0034 RBS and BBa_B0015 double terminator.

Source

The gene was extracted by pcr from Agrobacterium Tumefaciens gDNA.

http://www.ncbi.nlm.nih.gov/nucleotide/81176483?report=genbank&log$=nuclalign&blast_rank=1&RID=7BS4G4AC014.

The sequencing revealed mutations in our strain that do not affect the part functionality.

References