Difference between revisions of "Part:BBa K777000:Design"
(Undo revision 135711 by B.gene (Talk))
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===Design Notes===
 
===Design Notes===
 +
Genomic sequence was amplified using the following primers.<br><br>
 +
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´
 +
<br><br>
 +
<br><br>
 +
One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR. <br><br>
 +
Primers QuikChange: Primers were provided by SIGMA.
 +
 +
Fwd: TarQC for: 5´ ACTGATTGATTACCTAGATTATGGCAATACTGGAG 3´
 +
 +
Rev: TarQC rev: 5´ TGCCATAATCTAGGTAATCAATCAGTTCAGTTAAC 3´
  
 
===Source===
 
===Source===
  
 +
The part was amplified from genomic DNA of <i>E. coli</i> str. K-12 substr. DH10B chromosome (NC_010473.1).
  
 
===References===
 
===References===

Revision as of 07:51, 12 September 2012

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