Team:Cambridge/Oligos

Biobrick forward primer

>BBa_G00100 Part-only sequence (20 bp) tgccacctgacgtctaagaa

Biobrick backwards primer

>BBa_G00101 Part-only sequence (20 bp) attaccgcctttgagtgagc

The Vibrio Lux operon

AF170104-1

The Phosphoreum Lux operon

DQ988873

Pbad Promoter Part

Phosphoreum LuxL gene upstream of LuxC

The phosphoreum oligos we ordered

Notes on alterations to the SLOCK pHK724 clone

• The sequence which SLOCK sent us is just the sequence which is found on NCBI (found by sequence alignment of the two sequences)
• We do not know exactly what is in the plasmid, maybe some information can be found in the references here
• From the annotated start of the lux ICDABEG operon from NCBI there are no restriction sites in conflict with the BioBrick standards.
• Editing ideas:
  • Remove I and G regions from the operon (are these useful?)
  • Attach suffices and prefixes
  • Is this sufficient to then put into a BioBrick? what Chassis should we use?
• A good summary of the lux operon with quality references
• How to design a RBS

Oligo Order for Vibrio Fischeri on 13.08.10

Oligo Order for Vibrio Fischeri on 17.08.12

Firefly Oligos to be ordered

• Mutagenesis Oligos to be ordered before DNA 2.0 comes through
• Oligos to biobrick our DNA 2.0 order

Our first attempts at oligo design (a little excessive)

Oligos to BioBrick the whole lux operon (never ordered as too expensive)

luxICDABE operon sequence from NCBI

LuxR sequence from NCBI

Thioesterase

ygcB gene from E. Coli K12 to over-express thioesterase: GTGAATACAACGCTGTTTCGATGGCCGGTTCGCGTCTACTATGAAGATACCGATGCCGGTGGTGTGGTGTACCACGCCAGTTACGTCGCTTTTTATGAAA GAGCACGCACAGAGATGCTGCGTCATCATCACTTCAGTCAGCAGGCGCTGATGGCTGAACGCGTTGCCTTTGTGGTACGTAAAATGACGGTGGAATATTA CGCACCTGCGCGGCTCGACGATATGCTCGAAATACAGACTGAAATAACATCAATGCGTGGCACCTCTTTGGTTTTCACGCAACGTATTGTCAACGCCGAG AATACTTTGCTGAATGAAGCAGAGGTTCTGGTTGTTTGCGTTGACCCACTCAAAATGAAGCCTCGTGCGCTTCCCAAGTCTATTGTCGCGGAGTTTAAGC AGTGA

Forward Primer:

gaattcgcggccgcttctagaGTGAATACAACGCTGTTTC

Complement (for Gibson):

GAAACAGCGTTGTATTCACtctagaagcggccgcgaattc

Bacward primer:

ctgcagcggccgctactagtaTCACTGCTTAAACTCCG

Complement:

CGGAGTTTAAGCAGTGAtactagtagcggccgctgcag