Difference between revisions of "Part:BBa K2560257:Design"

(→‎Design Notes)
(→‎References)
Line 24: Line 24:
  
 
===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 21:41, 29 September 2018


4x Streptag for N-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

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 BBa_K2560258.

GGTCTCGAATG-coding_region-GATGTGAGACC

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 have a small linker to the 3' coding region, a GGG (glycine) was added as well as an ATG (methionine) as startcodon.

The part was codonoptimized for V. natriegens and then synthetisized by IDT and integrated into the vector BBa_K2560002 via 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