Difference between revisions of "Part:BBa K2560258:Design"

 
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===Design Notes===
 
===Design Notes===
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===Design Notes===
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This part is flanked by overhangs which are Phytobrick- and MoClo-compatible and by two BsaI recognition sites (Weber et al., 2011).  
−
To have a small linker to the 3' coding region, a GGG (glycine) was added as well as an ATG (methionine) as startcodon. This region is flanked by overhangs which are Phytobrick- and MoClo-compatible and by two BsaI recognition sites (Weber et al., 2011).  
+
  
−
If the Streptag shall be fused to the 5' end (N-terminus of the protein), use this part, for 3' fusion (C-terminus of the protein) use [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560258 BBa_K2560258].
+
If the Streptag shall be fused to the 3' end (C-terminus of the protein), use this part, for 5' fusion (N-terminus of the protein) use [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560257 BBa_K2560257].
  
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GGTCTCG<b>AATG</b>-coding_region-<b>GATG</b>TGAGACC
+
GGTCTCG<b>GCTT</b>TA-coding_region-G<b>GGTA</b>TGAGACC
  
 
The sequence was codonoptimized for <i>V. natriegens</i> ATCC 14048.
 
The sequence was codonoptimized for <i>V. natriegens</i> ATCC 14048.
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−
 
−
 
−
 
−
===Source===
 
  
 
===Source===
 
===Source===
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Source of the part:
 
Source of the part:
  
−
The sequence of this part was used from Addgene (Plasmid #55181). To have a small linker to the 3' coding region, a GGG (glycine) was added as well as an ATG (methionine) as startcodon.
+
The sequence of this part was used from Addgene (Plasmid #55181). To stay in frame, a TA was added before the coding region and a G behind it. (LWSHPQFEKG)
−
 
+
−
The part was codonoptimized for <i>V. natriegens</i> and then synthetisized by IDT and integrated into the vector BBa_K2560002 via BsmBI
+
  
 +
The part was codonoptimized for <i>V. natriegens</i> and then synthetisized by IDT and integrated into the vector [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560002 BBa_K2560002]via BsmBI
  
 
===References===
 
===References===
 +
Weber E., Engler C., Gruetzner R., Werner S., Marillonnet S. (2011). A modular cloning system for standardized assembly of multigene constructs. PLoS One 6:e16765. 10.1371/journal.pone.0016765

Revision as of 21:58, 29 September 2018


Streptag for C-terminal protein tagging


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

This part is flanked by overhangs which are Phytobrick- and MoClo-compatible and by two BsaI recognition sites (Weber et al., 2011).

If the Streptag shall be fused to the 3' end (C-terminus of the protein), use this part, for 5' fusion (N-terminus of the protein) use BBa_K2560257.

GGTCTCGGCTTTA-coding_region-GGGTATGAGACC

The sequence was codonoptimized for V. natriegens ATCC 14048.

Source

Source of the part:

The sequence of this part was used from Addgene (Plasmid #55181). To stay in frame, a TA was added before the coding region and a G behind it. (LWSHPQFEKG)

The part was codonoptimized for V. natriegens and then synthetisized by IDT and integrated into the vector BBa_K2560002via BsmBI

References

Weber E., Engler C., Gruetzner R., Werner S., Marillonnet S. (2011). A modular cloning system for standardized assembly of multigene constructs. PLoS One 6:e16765. 10.1371/journal.pone.0016765