Coding

Part:BBa_K4770031

Designed by: Andrea Camí Bonet   Group: iGEM23_Barcelona-UB   (2023-10-12)

LOG_Lonely_Guy_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: LOG

With this vector, we aim to see the effects of overexpressing LOG 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, LOG 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 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 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 4191
    Illegal PstI site found at 6531
    Illegal PstI site found at 8057
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 4191
    Illegal PstI site found at 6531
    Illegal PstI site found at 8057
    Illegal NotI site found at 7793
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4523
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 4191
    Illegal PstI site found at 6531
    Illegal PstI site found at 8057
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 4191
    Illegal PstI site found at 6531
    Illegal PstI site found at 8057
    Illegal NgoMIV site found at 1265
    Illegal NgoMIV site found at 1447
    Illegal NgoMIV site found at 4339
    Illegal NgoMIV site found at 8534
    Illegal AgeI site found at 3195
  • 1000
    COMPATIBLE WITH RFC[1000]
[edit]
Categories
Parameters
None