Difference between revisions of "Part:BBa S03595:Design"

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===Design Notes===
 
===Design Notes===
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Promoterless tetracycline resistance coding region (forward orientation) with a ribosomal binding site ('''RBS-TetF''') is expressed (tet resistant) in pSB1A2 and pSB1A3. See [https://parts.igem.org/wiki/index.php/Part:BBa_J31009:Design pSB1A4 Part Design] for details. This part was constructed to test the capacity of the double forward terminator [https://parts.igem.org/wiki/index.php/Part:BBa_B0015 BBa_B0015] to insulate a BioBrick part from read-through transcription coming from the vector backbone.  
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Promoterless tetracycline resistance coding region (forward orientation) with a ribosomal binding site ('''RBS-TetF''') is expressed (tet resistant) in pSB1A2 and pSB1A3 (see[https://parts.igem.org/wiki/index.php/Part:BBa_J31009:Design pSB1A4 Part Design] for details). This part was constructed to test the capacity of the double forward terminator [https://parts.igem.org/wiki/index.php/Part:BBa_B0015 BBa_B0015] to insulate a BioBrick part from read-through transcription coming from the vector backbone.  
  
 
RBS-TetF [https://parts.igem.org/wiki/index.php/Part:BBa_S03567 BBa_S03567] was placed downstream of the double forward terminator [https://parts.igem.org/wiki/index.php/Part:BBa_B0015 BBa_B0015] (TT) in pSB1AK3. Then TT-RBS-TetF (EcoRI/ PstI) was cloned into pSB1A2. When the TT is upstream of RBS-TetF, the cells are no longer tetracycline resistant. We conclude that the double forward terminator is sufficient to block read-through transcription. Thus, the double forward terminator was used to contruct a new cloning vector, '''pSB1A4''' ([https://parts.igem.org/wiki/index.php/Part:BBa_J31009 BBa_J31009]).
 
RBS-TetF [https://parts.igem.org/wiki/index.php/Part:BBa_S03567 BBa_S03567] was placed downstream of the double forward terminator [https://parts.igem.org/wiki/index.php/Part:BBa_B0015 BBa_B0015] (TT) in pSB1AK3. Then TT-RBS-TetF (EcoRI/ PstI) was cloned into pSB1A2. When the TT is upstream of RBS-TetF, the cells are no longer tetracycline resistant. We conclude that the double forward terminator is sufficient to block read-through transcription. Thus, the double forward terminator was used to contruct a new cloning vector, '''pSB1A4''' ([https://parts.igem.org/wiki/index.php/Part:BBa_J31009 BBa_J31009]).

Revision as of 20:36, 19 October 2006


TT : RBS-TetF


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 302
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 448
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 474
    Illegal NgoMIV site found at 842
    Illegal NgoMIV site found at 1002
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Promoterless tetracycline resistance coding region (forward orientation) with a ribosomal binding site (RBS-TetF) is expressed (tet resistant) in pSB1A2 and pSB1A3 (seepSB1A4 Part Design for details). This part was constructed to test the capacity of the double forward terminator BBa_B0015 to insulate a BioBrick part from read-through transcription coming from the vector backbone.

RBS-TetF BBa_S03567 was placed downstream of the double forward terminator BBa_B0015 (TT) in pSB1AK3. Then TT-RBS-TetF (EcoRI/ PstI) was cloned into pSB1A2. When the TT is upstream of RBS-TetF, the cells are no longer tetracycline resistant. We conclude that the double forward terminator is sufficient to block read-through transcription. Thus, the double forward terminator was used to contruct a new cloning vector, pSB1A4 (BBa_J31009).

Source

References