Coding

Part:BBa_K4171000

Designed by: HUANG, YEN-JUNG   Group: iGEM22_NCKU_Tainan   (2022-07-21)


CysK

Background

CysK is one of the members in the cysteine synthesis pathway in E. coli. Its acetylserinyl-sulfhydrylase activity catalyzes hydrogen sulfide and O-acetyl-L-serine (OAS) to form L-cysteine and acetate.

Usage

Since sulfur and selenium share lots of similar properties, we cultured E. coli with sodium selenite to replace sulfide and tested whether the enzymes that originally produce cysteine can also synthesize selenocysteine (Sec) as well. CysK (BBa_K4171000) was overexpressed in E. coli to convert sodium selenite and OAS into Sec.



Fig. 1. In vivo Sec synthesis pathway

Characterization



Fig. 2. Confirmation of cysK (BBa_K4171000) by PCR. M: Marker; Lane 1: cysK (972bp).


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 469
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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