Difference between revisions of "Part:BBa K525710:Design"

(→‎Design Notes)
(→‎Source)
 
(One intermediate revision by the same user not shown)
Line 11: Line 11:
 
===Source===
 
===Source===
  
−
PCR on the ''Escherichia coli'' TOP 10 DNA cloned into BioBrick Vectors
+
*PCR on the ''Escherichia coli'' TOP 10 DNA cloned into BioBrick Vectors
  
−
    Primers with BioBrick prefix, pT7-Promoter and RBS in the fwd primer. Suffix in the rev primer.
+
*Primers with BioBrick prefix, pT7-Promoter and RBS in the fwd primer. Suffix and His-tag in the rev primer.
−
        fwd:  
+
**fwd: 5'-acgtgaattcgcggccgcttctagagtaatacgactcactatagggaaagaggagaaaaatggaatcaatcgaacaac-3'
−
        rev:
+
**rev: 5'-acgtctgcagcggccgctactagtaatgatgatgatgatgatggctacccagcaaacgc-3'
  
 
===References===
 
===References===

Latest revision as of 20:24, 21 September 2011

DNA ligase from Escherichia coli (LigA) with PT7, RBS and His-tag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1449
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1663


Design Notes

This is the complete sequence of NAD+ -dependent DNA ligase with PT7, RBS and His-tag

Source

  • PCR on the Escherichia coli TOP 10 DNA cloned into BioBrick Vectors
  • Primers with BioBrick prefix, pT7-Promoter and RBS in the fwd primer. Suffix and His-tag in the rev primer.
    • fwd: 5'-acgtgaattcgcggccgcttctagagtaatacgactcactatagggaaagaggagaaaaatggaatcaatcgaacaac-3'
    • rev: 5'-acgtctgcagcggccgctactagtaatgatgatgatgatgatggctacccagcaaacgc-3'

References