Difference between revisions of "Part:BBa K2442403"
Line 9: | Line 9: | ||
This is a composite of the GFP N-term split subunit (GFP1) [https://parts.igem.org/Part:BBa_K1789003 K1789003] and the well characterised double-terminator construct [https://parts.igem.org/Part:BBa_B0015 B0015]. | This is a composite of the GFP N-term split subunit (GFP1) [https://parts.igem.org/Part:BBa_K1789003 K1789003] and the well characterised double-terminator construct [https://parts.igem.org/Part:BBa_B0015 B0015]. | ||
+ | |||
+ | |||
+ | This N-term GFP part was found to contain an extra A nucleotide immediately preceding the start-codon ATG. This erroneous nucleotide is present in the parts-registry sequencing for K1789003, but is not reported as an error. Unfortunately, now both split-GFP modules might be affected by improperly spaced ribosome binding sites. Despite this we proceeded to the next assembly stage. | ||
+ | |||
+ | [[Image:Glasgow_ANDgate_Figure_4.png|450px|thumb|center|A Biobrick prefix error in GFP N-term part.</b> Alignment of a sequencing read of the split GFP N-terminal to a reference sequence plasmid map. The base highlighted in red indicate an insertion has occurred at those positions relative to the reference sequence. The single-A insertion may correspond to the use a taq-polymerase during the original amplification of this coding sequence. Alignments performed in ApE.]] | ||
+ | |||
Revision as of 03:19, 2 November 2017
GFP split N-term (GFP2) + B0015 double terminator
This is a composite of the GFP N-term split subunit (GFP1) K1789003 and the well characterised double-terminator construct B0015.
This N-term GFP part was found to contain an extra A nucleotide immediately preceding the start-codon ATG. This erroneous nucleotide is present in the parts-registry sequencing for K1789003, but is not reported as an error. Unfortunately, now both split-GFP modules might be affected by improperly spaced ribosome binding sites. Despite this we proceeded to the next assembly stage.
![](/wiki/images/8/81/Glasgow_ANDgate_Figure_4.png)
A Biobrick prefix error in GFP N-term part.</b> Alignment of a sequencing read of the split GFP N-terminal to a reference sequence plasmid map. The base highlighted in red indicate an insertion has occurred at those positions relative to the reference sequence. The single-A insertion may correspond to the use a taq-polymerase during the original amplification of this coding sequence. Alignments performed in ApE.
Usage and Biology
this composite was used for assembly of larger composite split-GFP AND-gate constructs.
- K2442405 [split GFP module 1] tetR repressible promoter + GFP N-term + RBS + double terminator
- K2442408 Full split-GFP AND-gate circuit [Split GFP module 1 + Split GFP module 2]
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]