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Team:Cambridge/Oligos
From 2010.igem.org
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(→Notes on alterations to the SLOCK pHK724 clone) |
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[http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=BBa%20I0500 Pbad Promoter Part] | [http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=BBa%20I0500 Pbad Promoter Part] | ||
| + | |||
[http://www.fasebj.org/cgi/reprint/7/11/1016 Phosphoreum LuxL gene upstream of LuxC] | [http://www.fasebj.org/cgi/reprint/7/11/1016 Phosphoreum LuxL gene upstream of LuxC] | ||
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[[Team:Cambridge/OligoOrderVibrio13.08.10 | Oligo Order for Vibrio Fischeri on 13.08.10]] | [[Team:Cambridge/OligoOrderVibrio13.08.10 | Oligo Order for Vibrio Fischeri on 13.08.10]] | ||
| + | |||
| + | [[Team:Cambridge/OligoOrderVibrio17.08.10 | Oligo Order for Vibrio Fischeri on 17.08.12]] | ||
===Firefly Oligos to be ordered=== | ===Firefly Oligos to be ordered=== | ||
| - | [[Team:Cambridge/mutagenesisprimers17.08.10 | Mutagenesis Oligos to be ordered before DNA 2.0 comes through]] | + | *[[Team:Cambridge/mutagenesisprimers17.08.10 | Mutagenesis Oligos to be ordered before DNA 2.0 comes through]] |
| - | [[Team:Cambridge/biobrickingoligosfirefly | Oligos to biobrick our DNA 2.0 order]] | + | *[[Team:Cambridge/biobrickingoligosfirefly | Oligos to biobrick our DNA 2.0 order]] |
===Our first attempts at oligo design (a little excessive)=== | ===Our first attempts at oligo design (a little excessive)=== | ||
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[[Team:Cambridge/Biobrickingluxoperon | Oligos to BioBrick the whole lux operon]] (never ordered as too expensive) | [[Team:Cambridge/Biobrickingluxoperon | Oligos to BioBrick the whole lux operon]] (never ordered as too expensive) | ||
| - | [[Team:Cambridge/luxICDABEoperon | luxICDABE operon sequence]] | + | [[Team:Cambridge/luxICDABEoperon | luxICDABE operon sequence]] from NCBI |
[[Team:Cambridge/LuxR | LuxR sequence]] from NCBI | [[Team:Cambridge/LuxR | LuxR sequence]] from NCBI | ||
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AATACTTTGCTGAATGAAGCAGAGGTTCTGGTTGTTTGCGTTGACCCACTCAAAATGAAGCCTCGTGCGCTTCCCAAGTCTATTGTCGCGGAGTTTAAGC | AATACTTTGCTGAATGAAGCAGAGGTTCTGGTTGTTTGCGTTGACCCACTCAAAATGAAGCCTCGTGCGCTTCCCAAGTCTATTGTCGCGGAGTTTAAGC | ||
AGTGA | AGTGA | ||
| + | |||
| + | Forward Primer: | ||
| + | |||
| + | gaattcgcggccgcttctagaGTGAATACAACGCTGTTTC | ||
| + | |||
| + | Complement (for Gibson): | ||
| + | |||
| + | GAAACAGCGTTGTATTCACtctagaagcggccgcgaattc | ||
| + | |||
| + | Bacward primer: | ||
| + | |||
| + | ctgcagcggccgctactagtaTCACTGCTTAAACTCCG | ||
| + | |||
| + | Complement: | ||
| + | |||
| + | CGGAGTTTAAGCAGTGAtactagtagcggccgctgcag | ||
Latest revision as of 10:27, 18 August 2010
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
The Phosphoreum Lux operon
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
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
