Part:BBa_K2138000:Design
RNA thermometer and RBS (40C)
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 118
Illegal EcoRI site found at 2166
Illegal XbaI site found at 2181 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal EcoRI site found at 2166
Illegal NheI site found at 30
Illegal NheI site found at 53
Illegal SpeI site found at 119
Illegal PstI site found at 133
Illegal NotI site found at 7
Illegal NotI site found at 126
Illegal NotI site found at 2172 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
Illegal EcoRI site found at 2166
Illegal XhoI site found at 1150
Illegal XhoI site found at 2042 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 119
Illegal EcoRI site found at 2166
Illegal XbaI site found at 2181 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal EcoRI site found at 2166
Illegal XbaI site found at 16
Illegal XbaI site found at 2181
Illegal SpeI site found at 119
Illegal PstI site found at 133 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The primary design consideration while designing the DNA sequence was to achieve a melting temperature between thirty degrees Celsius and forty-two degrees Celsius. This was achieved by eliminating as many cytosine and guanine complementary bonds as possible leading to sequences comprised essentially of three different nucleotides.
Source
The DNA sequence is comprised of two different segments. The first segment was designed entirely de novo by utilizing Oligoanalyzer3.1 from Integrated DNA Technologies to calculate melting temperature for the complementary RNA strand. The second segment consists of the ribosome binding site which was selected based upon the Anderson ribosome binding site format for strong protein expression.