Plasmid
Part:BBa_K1321304:Design
Designed by: Michael Florea Group: iGEM14_Imperial (2014-10-08)
pSEVA321-BB with J23117-RFP
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 4476
Illegal SpeI site found at 37
Illegal PstI site found at 920 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4476
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal SpeI site found at 37
Illegal PstI site found at 920
Illegal NotI site found at 913
Illegal NotI site found at 4482 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4476
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 4476
Illegal SpeI site found at 37
Illegal PstI site found at 920 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 4476
Illegal XbaI site found at 4491
Illegal SpeI site found at 37
Illegal PstI site found at 920
Illegal NgoMIV site found at 2095
Illegal NgoMIV site found at 2980
Illegal NgoMIV site found at 4091
Illegal NgoMIV site found at 4215
Illegal AgeI site found at 616
Illegal AgeI site found at 728 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
BBa_K1321302 was created by restricting BBa_J23100 (Anderson promoter+RFP insert) and BBa_K1321300 (pSEVA331-BB backbone) with XbaI and PstI, gel purifying the resulting fragments and ligating with T4 ligase. Ligated DNA was then transformed into chemically competent cells, screened via colony PCR and culture PCR, and confirmed by sequencing.
Source
The original pSEVA321 backbone comes from the SEVA (Standard European Vector Architecture) collection.