Coding

Part:BBa_K4770032

Designed by: Andrea Camí Bonet   Group: iGEM23_Barcelona-UB   (2023-10-12)
Revision as of 12:23, 12 October 2023 by Andreacami (Talk | contribs)

NR_NiR_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: NR + pCM1 NiR With this vector, we aim to see the effects of overexpressing the first two genes of the nitrates assimilation pathway constitutively and simultaneously. 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), 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 assembled with HygroR cassette in position 3, pCM1 NR in position4, pCM1NiR 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 NR: BBa_K4770018 Position 5: pCM1 NiR BBa_K4770019 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.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 3882
    Illegal PstI site found at 5146
    Illegal PstI site found at 12173
    Illegal PstI site found at 13699
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 5726
    Illegal NheI site found at 7969
    Illegal PstI site found at 3882
    Illegal PstI site found at 5146
    Illegal PstI site found at 12173
    Illegal PstI site found at 13699
    Illegal NotI site found at 13435
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 6281
    Illegal BglII site found at 9699
    Illegal BglII site found at 10165
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 3882
    Illegal PstI site found at 5146
    Illegal PstI site found at 12173
    Illegal PstI site found at 13699
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 3882
    Illegal PstI site found at 5146
    Illegal PstI site found at 12173
    Illegal PstI site found at 13699
    Illegal NgoMIV site found at 1265
    Illegal NgoMIV site found at 1447
    Illegal NgoMIV site found at 3771
    Illegal NgoMIV site found at 4365
    Illegal NgoMIV site found at 4980
    Illegal NgoMIV site found at 6097
    Illegal NgoMIV site found at 8444
    Illegal AgeI site found at 3543
    Illegal AgeI site found at 5856
    Illegal AgeI site found at 9488
  • 1000
    COMPATIBLE WITH RFC[1000]
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Categories
Parameters
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