Develop Assembly Schedule


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Malcolm has put together a ppt that shows the latest, greatest plan for two different one pancake constructs and a four pancake construct. You may find it on the Davidson WIKI, or you can IM him and ask him to send it to you.

Here is a brief description of the construct:

One pancake constructs:

  1. lac promoter, RBS, Hin CDS, double terminator, hixC, normal or reversed pBAD (arabinose promoter), hixC, RBS, Tet resistance CDS, double terminator, RE
  2. lac promoter, RBS, Hin CDS, double terminator, normal pBAD, hixC, RBS, Tet resistance CDS, hixC, double terminator, RE

Here is a description of the four pancake construct:

lac promoter, RBS, Hin CDS, double terminator, hixC, (four pancakes to be flipped), double terminator, RE

The four pancakes to be flipped will come from this list of eight pancakes that we will make:

  1. pBAD - hixC
  2. reverse pBAD - hixC
  3. RBS - Tet resistance - hixC
  4. reversed (RBS - Tet resistance) - hixC
  5. Kan resistance - hixC
  6. reversed Kan resistance - hixC
  7. Chl resistance- double terminator - hixC
  8. reversed (Chl resistance- double terminator) - hixC

Note: we will have to make reversed RBS and reversed double terminator. Trevor - see if you can design PCR primers for this the way we did for the reversed pAra on Friday.

What we need is the following:

  1. complete list of biobricks available and being constructed, with registry numbers and DNA plate locations
  2. schedule of construction of new biobricks
  3. schedule with day one, two three etc. of which bricks will be put together first, then next, then next
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