Difference between revisions of "Part:BBa K653001:Design"

 
(→‎Source)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K653001 short</partinfo>
 
<partinfo>BBa_K653001 short</partinfo>
Line 20: Line 19:
 
===Source===
 
===Source===
  
−
atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtg
+
This BBa_E0040 part was derived from the reporter device used by Elowitz's repressilator [1]. Elowitz obtained the GFP tagged sequence (gfpmut3*) published by Andersen and coworkers [2] to build the reporter device. Andersen constructed those GFP variants based on a gfpmut3b containing plasmid that was a gift from R. H. Valdivia [3]. The PCR method used in construction by Andersen changed aa residue 2 from serine to arginine with no detectable phenotypic effect in their experiments.
−
aaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatgg
+
 
−
tgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttc
+
The BBa_B0015 Double Terminator part is the most commonly used terminator. It seems to be reliable.
−
aaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatg
+
Note, however, that Part:BBa_B0014 is a better part for forward and reverse termination. The part was designed by Reshma Shetty
−
gaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaat
+
−
tagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattac
+
−
ctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatgg
+
−
atgaactatacaaataataa ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctc
+
−
accttcgggtgggcctttctgcgtttata
+
  
 
===References===
 
===References===

Revision as of 22:45, 28 September 2011

GFP reporter plus Double Terminator


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 BsaI.rc site found at 644


Design Notes

This GFP part was derived from the reporter device used by Elowitz's repressilator : GFPmut3* has a couple of mutations from wtGFP - S2R and S65G and S72A. Emerald and EGFP have S65T and multiple other mutations. The S2R mutation should have no effect.

The Double terminator consisting of BBa_B0010 and BBa_B0012. This is the most commonly used terminator. It seems to be reliable.

The assembling of this two parts was performed using the 3A Assembly Method.



Source

This BBa_E0040 part was derived from the reporter device used by Elowitz's repressilator [1]. Elowitz obtained the GFP tagged sequence (gfpmut3*) published by Andersen and coworkers [2] to build the reporter device. Andersen constructed those GFP variants based on a gfpmut3b containing plasmid that was a gift from R. H. Valdivia [3]. The PCR method used in construction by Andersen changed aa residue 2 from serine to arginine with no detectable phenotypic effect in their experiments.

The BBa_B0015 Double Terminator part is the most commonly used terminator. It seems to be reliable. 

Note, however, that Part:BBa_B0014 is a better part for forward and reverse termination. The part was designed by Reshma Shetty

References