Part:BBa_K4770034

IPT_LOG_Level_M

Description and Biology

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

In AlgaGenix we have designed what we call “gene-pairs Level Ms”. All of them follow the same structure: they contain the HygroR cassette, level 1 for two different genes (pCM1:NR + pCM1 NiR, pCM1 GS+ pCM1 Cp-GS or pCM1 IPT + pCM1 LOG), mVenus cassette, and mCherry cassette.

In this case, we are using the combination pCM1: IPT + pCM1 LOG

With this vector, we aim to see the effects of simultaneously overexpressing two of the crucial enzymes for cytokine synthesis. Additionally, we are also able to normalize the expression level depending on the insertion site of the vector (by measuring the signal of the mCherry cassette), and at the same time, we will be able to see the decrease of nitrates concentration in the media due to an enhancement of the nitrates assimilation pathway thanks to the mVenus cassette signal.

How can this part be used

This Level M is aimed to assemble with HygroR cassette in position 3, pCM1 IPT in position 4, pCM1 LOG 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 3: Hygro R cassette BBa_K4770016

Position 4: pCM1 IPT: BBa_K4770022

Position 5: pCM1 LOG BBa_K4770023

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 its own design considerations that can be looked up on the individual part’s pages.

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 8240
  Illegal PstI site found at 10580
  Illegal PstI site found at 12106
  
• 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 8240
  Illegal PstI site found at 10580
  Illegal PstI site found at 12106
  Illegal NotI site found at 3203
  Illegal NotI site found at 4836
  Illegal NotI site found at 11842
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 5567
  Illegal BglII site found at 8572
  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 8240
  Illegal PstI site found at 10580
  Illegal PstI site found at 12106
  
• 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 8240
  Illegal PstI site found at 10580
  Illegal PstI site found at 12106
  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
  Illegal AgeI site found at 7244
  
• 1000
  COMPATIBLE WITH RFC[1000]