Difference between revisions of "Part:BBa K1321337:Design"

(→‎Source)
(→‎Design Notes)
 
(4 intermediate revisions by the same user not shown)
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
OEPCR
+
The BioBrick [[Part:BBa_I746909]] was used as a template for overhang-primer PCR to add the RFC25 prefix and suffix onto the sfGFP coding region. Primers used were as follows:
  
 +
Forward primer: 5'-GAATTCGCGGCCGCTTCTAGATGGCCGGCcgtaaaggcgaagagctg-3'
  
 +
Reverse primer: 5'-CTGCAGCGGCCGCTACTAGTATTAACCGGTtttgtacagttcatccataccatg-3'
 +
 +
Capital letters are the RFC25 prefix/suffix overhang, lower-case letters are the portion of the primer initially bound to template. The final submitted part was confirmed by sequencing with the [https://parts.igem.org/Primers/Catalog verification primers] VF2 and VR.
  
 
===Source===
 
===Source===
  
−
Made from the existing part [[BBa_I746909]]
+
Made from the existing part [[Part:BBa_I746909]]
  
 
===References===
 
===References===
 
Origin of sfGFP sequence:
 
Origin of sfGFP sequence:
 
Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006
 
Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006

Latest revision as of 17:09, 12 October 2014


sfGFP in Freiburg format (RFC 25)


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 10


Design Notes

The BioBrick Part:BBa_I746909 was used as a template for overhang-primer PCR to add the RFC25 prefix and suffix onto the sfGFP coding region. Primers used were as follows:

Forward primer: 5'-GAATTCGCGGCCGCTTCTAGATGGCCGGCcgtaaaggcgaagagctg-3'

Reverse primer: 5'-CTGCAGCGGCCGCTACTAGTATTAACCGGTtttgtacagttcatccataccatg-3'

Capital letters are the RFC25 prefix/suffix overhang, lower-case letters are the portion of the primer initially bound to template. The final submitted part was confirmed by sequencing with the verification primers VF2 and VR.

Source

Made from the existing part Part:BBa_I746909

References

Origin of sfGFP sequence: Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006