Part:BBa_K5115038

ribozyme connected: MTA, Hpn, RcnR_C35L

Introduction

This composite part combines BBa_K5115035(ribozyme+RBS+MTA+stem-loop), BBa_K5115036(ribozyme+RBS+hpn+stem-loop)and BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop) . We introduced this ribozyme-assisted polycistronic co-expression system in [2022](https://2022.igem.wiki/fudan/parts). By inserting [ribozyme sequences](https://parts.igem.org/Part:BBa_K4765020) between CDSs in a polycistron, the RNA sequences of Twister ribozyme conduct self-cleaving, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons *in vivo*.

With this design, we achieve co-expression of [MTA](https://parts.igem.org/Part:BBa_K5115050), Hpn 没有它的basic, [RcnR C35L](https://parts.igem.org/Part:BBa_K5115000) at similar level. MTA is a protein that can bind with nickel ions to reduce its toxity to the *E.coli*. The Hpn is a protein that can sequester metals that accumulate internally to reduce nickel's toxity to the *E.coli*. RcnR C35L can regulate the nickel ion channel proteins in the cell membrane to tune the nickel ion transport rate.

Characterization

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 935
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 198
  
• 1000
  COMPATIBLE WITH RFC[1000]