Coding
Part:BBa_K1172901:Design
Designed by: Tore Bleckwehl Group: iGEM13_Bielefeld-Germany (2013-08-26)
Alanine racemase from ''E. coli''
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 331
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 255
Illegal BamHI site found at 957 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 373
Illegal AgeI site found at 673 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 130
Design Notes
Amplification with Gibson-primer:
- Amplification of the alanin racemase (alr):
pSB1C3_alr_fwd (41 bp)
5'-AATTCGCGGCCGCTTCTAGATGCAAGCGGCAACTGTTGT-3'
pSB1C3_alr_rev (41 bp)
5'-CTGCAGCGGCCGCTACTAGTATTAATCCACGTATTTCATCG-3'
- Amplification of the pSB1C3-Backbone:
pSB1C3_alr_suf (41 bp)
5'-CGATGAAATACGTGGATTAATACTAGTAGCGGCCGCTGCAG-3'
pSB1C3_alr_pre (41 bp)
5'-ACAACAGTTGCCGCTTGCATCTAGAAGCGGCCGCGAATT-3'
Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].
Source
- Genomic DNA Escherichia coli K-12
- Isolation of the whole E. coli genome, see [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Whole_Genome_Isolation here]