Difference between revisions of "Part:BBa K2442397"
Line 25: | Line 25: | ||
Despite the sequence error this part was used for assembly of larger composite split-GFP AND-gate constructs. | Despite the sequence error this part was used for assembly of larger composite split-GFP AND-gate constructs. | ||
− | [https://parts.igem.org/Part:BBa_K2442400 K2442400] B0032 + GFP split C-term | + | *[https://parts.igem.org/Part:BBa_K2442400 K2442400] B0032 + GFP split C-term |
− | [https://parts.igem.org/Part:BBa_K2442406 K2442406] [split GFP module 2] lacI regulated promoter + RBS + GFP C-term + double terminator | + | *[https://parts.igem.org/Part:BBa_K2442406 K2442406] [split GFP module 2] lacI regulated promoter + RBS + GFP C-term + double terminator |
− | [https://parts.igem.org/Part:BBa_K2442408 K2442408] Full split-GFP AND-gate circuit [Split GFP module 1 + Split GFP module 2] | + | *[https://parts.igem.org/Part:BBa_K2442408 K2442408] Full split-GFP AND-gate circuit [Split GFP module 1 + Split GFP module 2] |
Revision as of 02:55, 2 November 2017
GFP C-term split subunit (GFP2)
Design
We designed a PCR reaction to generate a C-terminal GFP2 coding sequence part from E0040 which would work alongside the existing N-terminal GFP1 part K1789003 made by NUDT_CHINA 2015. Primers were designed to amplify the coding sequence of the 83 aa C-terminal fragment as a biobrick compatible part.
Sequencing of the PCR for the GFP C-terminal part revealed an error – the PCR design had incorporated an incorrect version of the BioBrick prefix.
BioBrick parts beginning with ATG must use a shortened version of the prefix ending TAG, instead of the longer version for non-ATG parts which ends TAGAG. The short version ensures ideal spacing between upstream ribosome binding site parts and the start codon. Unfortunately, this oversight of the primer design was not initially noticed our plasmid map sequence for the C-terminal GFP part also contained the error. A part such as this with the extra two nucleotides before the start codon has the potential to be translated at decreased level from an upstream ribosome binding site.
Usage and Biology
Despite the sequence error this part was used for assembly of larger composite split-GFP AND-gate constructs.
- K2442400 B0032 + GFP split C-term
- K2442406 [split GFP module 2] lacI regulated promoter + RBS + GFP C-term + double terminator
- K2442408 Full split-GFP AND-gate circuit [Split GFP module 1 + Split GFP module 2]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 176