Coding

Part:BBa_K777000:Design

Designed by: Bianca Genenncher   Group: iGEM12_Goettingen   (2012-09-11)
Revision as of 20:01, 11 September 2012 by B.gene (Talk | contribs) (→‎Design Notes)

Tar receptor


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 1282
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 111


Design Notes

Genomic sequence was amplified using the following primers.

Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.

Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´

Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´



One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR.

Primers QuikChange: Primers were provided by SIGMA.

Fwd: TarQC for: 5´ ACTGATTGATTACCTAGATTATGGCAATACTGGAG 3´

Rev: TarQC rev: 5´ TGCCATAATCTAGGTAATCAATCAGTTCAGTTAAC 3´

Source

The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B chromosome (NC_010473.1).

References