Difference between revisions of "Part:BBa K1484001:Design"

 
(→‎Design Notes)
 
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===Design Notes===
 
===Design Notes===
−
The tsPurple chromoprotein is now compatible with both standards and can be used more effectively.
 
  
 +
tsPurple has been designed to be compatible with GoldenGate standards. This allows the part to be used in MoClo (ModularCloning) or GB2 (goldenbraid2) assembly method. These cloning techniques are based on based on Type IIS restriction enzymes that enable parallel assembly of multiple parts in a one-pot, one-step reaction and are becoming widely adopted across the field of Synthetic Biology.
  
 +
To be in the GoldenGate format, a level 0 CDS part must:
 +
start with
 +
-the BsaI_F recognition site
 +
  GGTCTC
 +
-one non-specific nucleotide
 +
  A
 +
-4bp fusion sequence (for CDS this is AATG)
 +
  AATG
 +
 +
finish with
 +
-4bp fusion sequence (for CDS this is    )
 +
  GCTT
 +
-one non-specific nucleotide
 +
  T
 +
-the BsaI_R recognition site reversed
 +
  GAGACC
 +
 +
 +
The tsPurple GoldenGate biobrick reads:
 +
Prefix_GGTCTC A AATG ___tsPurple(w/o atg)___GCTT T GAGACC_Suffix
  
 
===Source===
 
===Source===

Latest revision as of 08:26, 5 October 2014


tsPurple chromoprotein in GoldenGate standard


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 704


Design Notes

tsPurple has been designed to be compatible with GoldenGate standards. This allows the part to be used in MoClo (ModularCloning) or GB2 (goldenbraid2) assembly method. These cloning techniques are based on based on Type IIS restriction enzymes that enable parallel assembly of multiple parts in a one-pot, one-step reaction and are becoming widely adopted across the field of Synthetic Biology.

To be in the GoldenGate format, a level 0 CDS part must: start with -the BsaI_F recognition site 

  GGTCTC

-one non-specific nucleotide 

  A

-4bp fusion sequence (for CDS this is AATG) 

  AATG

finish with -4bp fusion sequence (for CDS this is ) 

  GCTT

-one non-specific nucleotide 

  T

-the BsaI_R recognition site reversed 

  GAGACC


The tsPurple GoldenGate biobrick reads: Prefix_GGTCTC A AATG ___tsPurple(w/o atg)___GCTT T GAGACC_Suffix

Source

The chromoprotein was taken from the iGEM registry and altered to conform to the MoClo standard.

References