Western Assembly Plan

From 2006.igem.org

Note: Western Team -- We need to send the backRBS part to Davidson as soon as we are confident that we have it for their Construct 3.

Parts List

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)

Upon receipt of the Hin and HixC from Davidson, Western will assemble the following Hin Expression Unit, both with and without an additional HixC, and send some of the finished product to Davidson (specifically that without the HixC site). Note: We need to build FOUR Hin cassettes -- with and without LVA for both teams, and with HixC for Western and without HixC for Davidson.

Western will also assemble Superparts necessary for the 1-pancake constructs in which the pAra promoter is both forward and backward. See diagram below.

Upon receipt of the TT-RE from Davidson, Western will construct Diamond 3 as below and complete final assembly for Construct 1 and Construct 1 – Control.