Davidson Assembly Plan
pLac promoter -- BBa_R0010 DNA-1 7K
pAra promoter -- BBa_I13453 DNA-2 13D
RBS -- BBa_B0030 DNA-1 3G
HixC – BBa_B?????
double forward terminator -- BBa_B0015 DNA-1 1I
double backward terminator – BBa_B0025 DNA-1 3E
KanR – BBa_B?????
Recombination enhancer -- BBa_J3101 (Davidson)
Hin -- BBa_J31000 (Davidson)
pAra promoter reverse – BBa_B????? (Western)
RBS reverse – BBa_B????? (Western)
Davidson will complete the TT-RE assembly below and send to Western.
The Davidson cohort will also construct SuperParts necessary for the 1-pancake constructs in which the Kan resistance coding region is both forward and backward. Note: Davidson will need to send the forward KanR to Western for use in their assembly.
Upon receipt of the Hin Expression Unit (no Hix) from Western, Davidson will assemble the following and complete the assembly of Construct 2 and Construct 2 – Control.
Construct 2 will allow us to assess if an RBS has the ability to reach across a HixC site and act on the CDS. We have reason to think that this may not be possible. Therefore, the Western Team will send the backRBS part to Davidson as soon as possible and the Davidson Team will build Constructs 3 below for which the RBS and CDS are both within the pancake.