Coding

Part:BBa_K1590008:Design

Designed by: Manuel Blank   Group: iGEM15_Dundee   (2015-09-09)


Lactoferrin Binding Protein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 172
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Biobrick Prefix, Suffix, and TAA stop codons had to be added. Sequence was checked for forbidden restriction sites in order to comply with assembly standard RFC10.

Forward primer for amplifying lactoferrin binding protein A from Neisseria meningitidis MC58 gDNA. 

  Forward: GCGC GAATTCGCGGCCGCTTCTAG A TGAATAAGAAACACGGTTTTCCG

  Reverse: GCGC CTGCAGCGGCCGCTACTAGT ATTATT AAAACTTCATTTCGAGCG

Intermediate sequencing primers for Lactoferrin Binding Protein. 

  Forward 1: GAAGTCGCTCGGCGCGGGG

  Forward 2: GACAAGAGCGGGGTGTACG

  Reverse  : GCCGAATCAGGCAGGAC

Primers for cloning Lactoferrin Binding Protein into overexpression vector pQE80-L. 

  Forward: GCGC GGATCC AATAAGAAACACGGTTTTCC

  Reverse: GCGC GGTACC TTAAAACTTCATTTCGAGCG


Source

PCR amplification of Neisseria Meningitidis MC58 gDNA. gDNA provided by James Moir - University of York.


References