Part:BBa_K4770030

IPT_Isopentenyltransferase_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:IPT

With this vector, we aim to see the effects of overexpressing IPT 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 due to a higher demand of nitrates assimilation to obtain the needed N for cytokines synthesis 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, IPT 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 IPT BBa_K4770022

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 PstI site found at 3323
  Illegal PstI site found at 4455
  Illegal PstI site found at 4876
  Illegal PstI site found at 4900
  Illegal PstI site found at 7575
  Illegal PstI site found at 9101
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 3323
  Illegal PstI site found at 4455
  Illegal PstI site found at 4876
  Illegal PstI site found at 4900
  Illegal PstI site found at 7575
  Illegal PstI site found at 9101
  Illegal NotI site found at 3203
  Illegal NotI site found at 4836
  Illegal NotI site found at 8837
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 5567
  Illegal BamHI site found at 4195
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 3323
  Illegal PstI site found at 4455
  Illegal PstI site found at 4876
  Illegal PstI site found at 4900
  Illegal PstI site found at 7575
  Illegal PstI site found at 9101
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 3323
  Illegal PstI site found at 4455
  Illegal PstI site found at 4876
  Illegal PstI site found at 4900
  Illegal PstI site found at 7575
  Illegal PstI site found at 9101
  Illegal NgoMIV site found at 1265
  Illegal NgoMIV site found at 1447
  Illegal NgoMIV site found at 3253
  Illegal NgoMIV site found at 4267
  Illegal NgoMIV site found at 4533
  Illegal NgoMIV site found at 4937
  Illegal NgoMIV site found at 4949
  
• 1000
  COMPATIBLE WITH RFC[1000]