Difference between revisions of "Part:BBa K1465228:Design"

(→‎Design Notes)
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
  
−
*Amplification of the pSB1C3-Backbone (<bbpart>BBa_J04450</bbpart>) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-pSB1C3-BBa_B0034-SBPase fw-pSB1C3-BBa_B0034-SBPase] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-SBPase-BBa_B0034-pSB1C3 rv-SBPase-BBa_B0034-pSB1C3]:<br>
+
* Amplification of the pSB1C3-Backbone (<bbpart>BBa_J04450</bbpart>) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-SBPase-pSB1C3 fw-SBPase-pSB1C3] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-SBPase-BBa_B0034-pSB1C3 rv-SBPase-BBa_B0034-pSB1C3]:<br>
  
−
  '''fw-pSB1C3-BBa_B0034-SBPase (58 bp)'''<br>
+
  '''fw-SBPase-pSB1C3 (57 bp)'''<br>
−
  5'-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3'<br>
+
  5'-AATTTATTGAAGCTTATCATTGATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAG-3'<br>
 
  '''rv-SBPase-BBa_B0034-pSB1C3 (65 bp)'''<br>
 
  '''rv-SBPase-BBa_B0034-pSB1C3 (65 bp)'''<br>
 
  5'-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3'<br>
 
  5'-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3'<br>
  
−
*Amplification of the front part of the Sedoheptulose-1,7-Bisphosphatase (''glpX'') for the removal of the illegal recostriction site. This part contains the RBS <bbpart>BBa_B0034</bbpart> and the front part of the Sedoheptulose-1,7-Bisphosphatase from ''Bacillus metanolicus''. For the amplification the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw_pSB1C3_csoS4A csoS4A_fwd] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv_csoS4A_PstI csoS4A_PstI-rev] were used:<br>
+
* Amplification of the front part of the Sedoheptulose-1,7-Bisphosphatase (''glpX'') for the removal of the illegal XbaI restriction site. This part contains the RBS <bbpart>BBa_B0034</bbpart> and the front part of the Sedoheptulose-1,7-Bisphosphatase from ''Bacillus metanolicus''. For the amplification the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-pSB1C3-BBa_B0034-SBPase fw-pSB1C3-BBa_B0034-SBPase] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-SBPase-Schnittstelle rv-SBPase-XbaI] were used:<br>
 +
 
 +
'''fw-pSB1C3-BBa_B0034-SBPase (58 bp)'''<br>
 +
5'-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3'<br>
 +
'''rv-SBPase-XbaI (46 bp)'''<br>
 +
5'-TTTTCTGTTAAAGATGCGTCCAAATTTACGCAGCCTTTTGCTTTTG-3'<br>
 +
 
 +
* Amplification of the back part of the Sedoheptulose-1,7-Bisphosphatase (''glpX'') for the removal of the illegal XbaI restriction site. For the amplification the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-SBPase-Schnittstelle fw-SBPase-XbaI] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-pSB1C3-SBPase rv-pSB1C3-SBPase] were used:<br>
 +
 
 +
'''fw-SBPase-XbaI (56 bp)'''<br>
 +
5'-AAAAGCAAAAGGCTGCGTAAATTTGGACGCATCTTTAACAGAAAATATGAAATCAG-3'<br>
 +
'''rv-pSB1C3-SBPase (56 bp)'''<br>
 +
5'-GACTGCAGCGGCCGCTACTAGTATCAATGATAAGCTTCAATAAATTGGTATTTTAC-3'<br>
 +
 
 +
*[http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone and the coding sequences of the Sedoheptulose-1,7-Bisphosphatase SBPase from ''Bacillus metanolicus'' containing also the RBS <bbpart>BBa_B0034</bbpart>.<br>
  
 
===Source===
 
===Source===

Revision as of 23:47, 8 October 2014


Sedoheptulose 1,7-bisphosphatase (glpX) from Bacillus methanolicus


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
    COMPATIBLE WITH RFC[1000]


Design Notes

  • Amplification of the pSB1C3-Backbone (BBa_J04450) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-SBPase-pSB1C3 fw-SBPase-pSB1C3] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-SBPase-BBa_B0034-pSB1C3 rv-SBPase-BBa_B0034-pSB1C3]:
fw-SBPase-pSB1C3 (57 bp)

5'-AATTTATTGAAGCTTATCATTGATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAG-3'

rv-SBPase-BBa_B0034-pSB1C3 (65 bp)

5'-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3'
  • Amplification of the front part of the Sedoheptulose-1,7-Bisphosphatase (glpX) for the removal of the illegal XbaI restriction site. This part contains the RBS BBa_B0034 and the front part of the Sedoheptulose-1,7-Bisphosphatase from Bacillus metanolicus. For the amplification the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-pSB1C3-BBa_B0034-SBPase fw-pSB1C3-BBa_B0034-SBPase] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-SBPase-Schnittstelle rv-SBPase-XbaI] were used:
fw-pSB1C3-BBa_B0034-SBPase (58 bp)

5'-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3'

rv-SBPase-XbaI (46 bp)

5'-TTTTCTGTTAAAGATGCGTCCAAATTTACGCAGCCTTTTGCTTTTG-3'
  • Amplification of the back part of the Sedoheptulose-1,7-Bisphosphatase (glpX) for the removal of the illegal XbaI restriction site. For the amplification the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#fw-SBPase-Schnittstelle fw-SBPase-XbaI] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#rv-pSB1C3-SBPase rv-pSB1C3-SBPase] were used:
fw-SBPase-XbaI (56 bp)

5'-AAAAGCAAAAGGCTGCGTAAATTTGGACGCATCTTTAACAGAAAATATGAAATCAG-3'

rv-pSB1C3-SBPase (56 bp)

5'-GACTGCAGCGGCCGCTACTAGTATCAATGATAAGCTTCAATAAATTGGTATTTTAC-3'
  • [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone and the coding sequences of the Sedoheptulose-1,7-Bisphosphatase SBPase from Bacillus metanolicus containing also the RBS BBa_B0034.

Source

Gibson

References