Coding

Part:BBa_K1590009:Design

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


Spermidine/putrescine-binding periplasmic protein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 429
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 148
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Adding biobrick prefix, suffix, and double TAA stop codon to it via primers. Checked for forbidden restriction sites.

Primers for amplification of PotD from E. coli MG1655 gDNA and subsequent insertion into pSB1C3. 

  Forward: GCGC GAATTCGCGGCCGCTTCTAG A TGAAAAAATGGTCACGCCAC

  Reverse: GCGC CTGCAGCGGCCGCTACTAGT ATTATT AACGTCCTGCTTTCAGCTT

Primers for cloning PotD into overexpression vector pQE80-L. 

  Forward: GCGC AGATCTAAAAAATGGTCACGCCAC

  Reverse: GCGC GGTACCTTAACGTCCTGCTTTCAGCTTC


Source

PCR amplification of Escherichia Coli MG1655 gDNA.

References