Difference between revisions of "Part:BBa K3866003"
(→‎Usage and Biology)
 
(4 intermediate revisions by the same user not shown)
Line 11: Line 11:
 
T5 promoter has been used for many years for protein expression in E.coli. Partial T5 promoter is used with the downstream Operator CuO.
 
T5 promoter has been used for many years for protein expression in E.coli. Partial T5 promoter is used with the downstream Operator CuO.
  
−
 
−
 
−
 
 
===Design Notes===
 
===Design Notes===
 
The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 <bbpart>BBa_K3505007</bbpart> and has overhangs compatible for GoldenBraid cloning.  
 
The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 <bbpart>BBa_K3505007</bbpart> and has overhangs compatible for GoldenBraid cloning.  
 
This has position A1-B2.
 
This has position A1-B2.
 
[[File:T--Thessaly--GB-GGAG-AATG.jpeg|700px|thumb|none|<i><b>Fig.1:</b>The overhangs of this part in the GoldenBraid Grammar</i>]]
 
[[File:T--Thessaly--GB-GGAG-AATG.jpeg|700px|thumb|none|<i><b>Fig.1:</b>The overhangs of this part in the GoldenBraid Grammar</i>]]
−
 
−
 
  
 
===Sequence and Features===
 
===Sequence and Features===

Latest revision as of 10:16, 15 October 2021


Partial T5 Promoter - CuO- RBS GB compatible with A1-B2

Partial T5 promoter is a strong constitutive Promoter that can be used in the toxin-antitoxin system with the Cym repressor that our team uses in their system.

Fig.2:Partial T5 Promoter with Lactose Operator and Cumate Operator with compatible ends for Golden Braid Cloning

Usage and Biology

T5 promoter has been used for many years for protein expression in E.coli. Partial T5 promoter is used with the downstream Operator CuO.

Design Notes

The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. This has position A1-B2.

Fig.1:The overhangs of this part in the GoldenBraid Grammar

Sequence and Features


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]

Source

Synthesized by Twist Biosciences.

References

  • Choi, Y. J., Morel, L., Le François, T., Bourque, D., Bourget, L., Groleau, D., Massie, B., & Míguez, C. B. (2010). Novel, versatile, and tightly regulated expression system for Escherichia coli strains. Applied and environmental microbiology, 76(15), 5058–5066. https://doi.org/10.1128/AEM.00413-10