Composite

Part:BBa_K3726054

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-19)
Revision as of 01:01, 20 October 2021 by Sbarraga (Talk | contribs)

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Lv.1_PKMCG-B

This is a composite part that corresponds with a level 1 MoClo part in accordance with the marburg collection standard. It is a transcriptional unit for the expression of the part “BBa_K3726028” L0_PKMCG which contains the half part of the MCG cycle and the enzymes that belong to the phosphoketolase pathway (X. Liu, R. Miao, P. Lindberg and P. Lindblad, "Modular engineering for efficient photosynthetic biosynthesis of 1-butanol from CO2in cyanobacteria", 2021).

The other half of the MCG cycle is going to be expressed in the part MCGII_Lv1 which has two variants “BBa_K3726055” L1_MCGII-A and “BBa_K3726056” L1_MCGII-B. The Malate-acetyl-CoA-Glycerate (MCG) cycle aims to the production of acetyl-CoA with high efficiency coupled to carbon capture from bicarbonate anion and with recovery of metabolic intermediates produced during photorespiration. (H. Yu, X. Li, F. Duchoud, D. Chuang and J. Liao, "Augmenting the Calvin–Benson–Bassham cycle by a synthetic malyl-CoA-glycerate carbon fixation pathway", 2021).

The part is flanked by two homology regions “ BBa_K3726112” 5CON1(H)_nNS5-up (PCC 11801) and “BBa_K3726113” 3CON5(H)_nNS5-down (PCC 11801) for the double homologous recombination within the genome of PCC 11801.

Likewise, a Spectinomycin resistance cassette is included downstream the transcriptional unit region.

Expression is controlled by the strong constitutive promoter for cyanobacteria, PS-PR “BBa_K3726084” and the strong synthetic RBS, RBS_PtaBs “BBa_K3726092”.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 9319
    Illegal SpeI site found at 10282
    Illegal PstI site found at 575
    Illegal PstI site found at 839
    Illegal PstI site found at 4205
    Illegal PstI site found at 4834
    Illegal PstI site found at 5331
    Illegal PstI site found at 5930
    Illegal PstI site found at 6868
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 9319
    Illegal NheI site found at 8971
    Illegal NheI site found at 8998
    Illegal SpeI site found at 10282
    Illegal PstI site found at 575
    Illegal PstI site found at 839
    Illegal PstI site found at 4205
    Illegal PstI site found at 4834
    Illegal PstI site found at 5331
    Illegal PstI site found at 5930
    Illegal PstI site found at 6868
    Illegal NotI site found at 9325
    Illegal NotI site found at 10289
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 9319
    Illegal BamHI site found at 505
    Illegal BamHI site found at 2744
    Illegal BamHI site found at 9864
    Illegal XhoI site found at 9195
    Illegal XhoI site found at 11313
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 9319
    Illegal suffix found in sequence at 10282
    Illegal PstI site found at 575
    Illegal PstI site found at 839
    Illegal PstI site found at 4205
    Illegal PstI site found at 4834
    Illegal PstI site found at 5331
    Illegal PstI site found at 5930
    Illegal PstI site found at 6868
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 9319
    Illegal XbaI site found at 9334
    Illegal SpeI site found at 10282
    Illegal PstI site found at 575
    Illegal PstI site found at 839
    Illegal PstI site found at 4205
    Illegal PstI site found at 4834
    Illegal PstI site found at 5331
    Illegal PstI site found at 5930
    Illegal PstI site found at 6868
    Illegal NgoMIV site found at 2321
    Illegal NgoMIV site found at 4430
    Illegal NgoMIV site found at 4890
    Illegal NgoMIV site found at 8786
    Illegal NgoMIV site found at 8928
    Illegal AgeI site found at 6443
    Illegal AgeI site found at 6526
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 9349
    Illegal BsaI.rc site found at 10266


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