Measurement

Part:BBa_J119127

Designed by: Todd Eckdahl   Group: Eckdahl Lab   (2012-10-29)

For Testing New Promoters via Golden Gate Assembly with BsmBI

This part allows users to clone and test new promoters without gel purification or other preparation of DNA. It is a destination vector for [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate Golden Gate Assembly] (GGA) using BsmBI and Ligase. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clone. The new promoter must be flanked by BsmBI sites that produce the 4 nt overhangs required for assembly (see Part:BBa_J119022 as an example, or the picture below). Successful GGA assembly replaces the double terminator in BBa_J119127 with the new promoter. Upon assembly, a functional new promoter should cause RFP expression. BBa_J119127 incorporates the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.

[http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Golden_Gate_Assembly_protocol See the full GGA protocol online.]


Sequence and Features


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 757
    Illegal AgeI site found at 869
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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