Coding
Part:BBa_K1590000:Design
Designed by: Manuel Blank Group: iGEM15_Dundee (2015-09-08)
Human Haemoglobin A
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 342
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Choosing the correct codon-bias, remove 'illegal' restriction sites, ensure that (stop)codons are in frame, addition of standard prefix and suffix.
Primers for cloning Haemoglobin A into overexpression vector pQE80-L.
Forward: 5' GCGC GGATCC GTTCTGTCTCCGGCGGAC 3' Reverse: 5' GCGC GGTACC TTACGGTATTTAGAGGT 3'
Primers for cloning Haemoglobin A into vector for bacterial 2-hybrid system, pT25.
Forward: 5' GCGC GGATCC AGTTCTGTCTCCGGCGGAC 3' Reverse: 5' GCGC GGTACC TTACGGGGTGAACTCCGCCGG 3'
Part was synthesised with appropriate prefix/suffix sequences, hence no separate primers were required for cloning it into pSB1C3.
Source
In silico optimised version for E. coli K12 strains. Sequence ordered through IDT.