Part:BBa_K4770028

GS_Glutamate_Synthetase_Level_M

Description and Biology

Level Ms are the assembly of different transcriptional units into the same vector.

All AlgaGenix’s Level M follow the same structure: they contain the HygroR cassette, level 1 for each gene (pCM1:NR, NiR, GS, Cp-GS, IPT or LOG), mVenus cassette, and mCherry cassette.

In this case, we are using pCM1:GS

With this vector, we aim to see the effects of overexpressing GS constitutively and at the same time normalizing the expression level depending on the insertion site of the vector (by measuring the signal of the mCherry cassette), at the same time, we will be able to see the decrease of nitrates concentration fue to higher assimilation thanks to the mVenus cassette signal.

How can this part be used

This Level M is aimed to assembled with HygroR cassette in position 4, GS Level1 in position 5, mCherry cassette in position 6 and mVenus cassette in position 7.

Should you use this part and need advice on how to use it, please do not hesitate to contact us at algagenix@gmail.com

Source of this part

Position 4: Hygro R cassette BBa_K4770016

Position 5: pCM1 GS BBa_K4770020

Position 6: mCherry cassette BBa_K4770017

Position 7: mVenus cassette BBa_K4770025

Design considerations

AlgaGenix’s Level Ms are designed to have the same structure and not having to build different pieces with different positions for each transcriptional unit. Each piece has it’s own design considerations that can be looked up on the individual part’s pages.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 4725
  Illegal PstI site found at 7071
  Illegal PstI site found at 8597
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 4725
  Illegal PstI site found at 7071
  Illegal PstI site found at 8597
  Illegal NotI site found at 8333
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 4725
  Illegal BglII site found at 5063
  Illegal BamHI site found at 4369
  Illegal XhoI site found at 4581
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 4725
  Illegal PstI site found at 7071
  Illegal PstI site found at 8597
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 4725
  Illegal PstI site found at 7071
  Illegal PstI site found at 8597
  Illegal NgoMIV site found at 1265
  Illegal NgoMIV site found at 1447
  Illegal NgoMIV site found at 3927
  Illegal NgoMIV site found at 4879
  Illegal NgoMIV site found at 9074
  
• 1000
  COMPATIBLE WITH RFC[1000]