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Welcome to your project page!
For each part listed, you should:
- Design oligos to make your part
- Write up proper construction files and put it on the Construction Files page
- Put your oligos on the Oligo Log page
- Put your part sequence on the part document
- Name your parts according to the list here
upaG Autotransporter long version
Source: CFT073 genomic DNA
First of all, read PMID: 18424525. Also download this template for your design here. You are going to make the part that includes the "Him region" (blue) and the "L1 L2 beta" region (red).
Due to the large size, you won't sequence every base of your part. You should do a reverse read with G00101 to confirm that the "L1 L2 beta" region is correct in your part, and you should also do a forward ca998 read over the N-terminus. Ignore the middle.
This part encodes a N-terminal display protein
Your displayer part should be of the {<part!} style (no start, with stop). There should be NO prepro sequence in your part. You should design your construction file to insert your part into plasmid pBca9495KC-Bca1144#5 using EcoRI and BamHI. The map of this plasmid is here.
Alkaline Phosphatase
Source: pBca9145-Bca1126 {<phoA>}
The oligos to construct this part have already been designed. You just need to complete the construction file below. As background, you should read PMID: 7929016. The map of the template sequence is here.
PCR mea036/mea037 on pBca9145-Bca1126 (1219 bp, EcoRI/BamHI)
Sub into ????? (EcoRI/BamHI, ???, ???)
Product is ????
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mea036 Forward Biobricking of {<phoA>} cgatagaattcatgAGATCTGGAGATTCTG
mea037 Reverse Biobricking of {<phoA>} cgttaGGATCCCTTCAGGCCCAGCGCCGCTTTCATG
This part encodes a passenger protein
Your passenger part should be of the {<part>} style (no start, no stop). There should be NO prepro sequence in your part. If your part is naturally secreted, you should encode only the active peptide. You should design your construction file to insert your part into plasmid pBca9495AK-Bca1144#5 using EcoRI and BamHI. The map of this plasmid is here.
