Composite

Part:BBa_K3726056:Design

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-19)
Revision as of 01:16, 20 October 2021 by Sbarraga (Talk | contribs)


Lv.1_PKMCG-B


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2173
    Illegal EcoRI site found at 2614
    Illegal EcoRI site found at 4373
    Illegal EcoRI site found at 5489
    Illegal EcoRI site found at 5573
    Illegal EcoRI site found at 8628
    Illegal PstI site found at 2758
    Illegal PstI site found at 2857
    Illegal PstI site found at 3442
    Illegal PstI site found at 4400
    Illegal PstI site found at 4628
    Illegal PstI site found at 6138
    Illegal PstI site found at 7152
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2173
    Illegal EcoRI site found at 2614
    Illegal EcoRI site found at 4373
    Illegal EcoRI site found at 5489
    Illegal EcoRI site found at 5573
    Illegal EcoRI site found at 8628
    Illegal NheI site found at 5712
    Illegal PstI site found at 2758
    Illegal PstI site found at 2857
    Illegal PstI site found at 3442
    Illegal PstI site found at 4400
    Illegal PstI site found at 4628
    Illegal PstI site found at 6138
    Illegal PstI site found at 7152
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2173
    Illegal EcoRI site found at 2614
    Illegal EcoRI site found at 4373
    Illegal EcoRI site found at 5489
    Illegal EcoRI site found at 5573
    Illegal EcoRI site found at 8628
    Illegal BglII site found at 1115
    Illegal BglII site found at 3155
    Illegal BglII site found at 4519
    Illegal BamHI site found at 3253
    Illegal BamHI site found at 3268
    Illegal BamHI site found at 6967
    Illegal BamHI site found at 7425
    Illegal BamHI site found at 8398
    Illegal XhoI site found at 337
    Illegal XhoI site found at 1945
    Illegal XhoI site found at 2590
    Illegal XhoI site found at 3064
    Illegal XhoI site found at 5594
    Illegal XhoI site found at 6261
    Illegal XhoI site found at 6394
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2173
    Illegal EcoRI site found at 2614
    Illegal EcoRI site found at 4373
    Illegal EcoRI site found at 5489
    Illegal EcoRI site found at 5573
    Illegal EcoRI site found at 8628
    Illegal PstI site found at 2758
    Illegal PstI site found at 2857
    Illegal PstI site found at 3442
    Illegal PstI site found at 4400
    Illegal PstI site found at 4628
    Illegal PstI site found at 6138
    Illegal PstI site found at 7152
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2173
    Illegal EcoRI site found at 2614
    Illegal EcoRI site found at 4373
    Illegal EcoRI site found at 5489
    Illegal EcoRI site found at 5573
    Illegal EcoRI site found at 8628
    Illegal PstI site found at 2758
    Illegal PstI site found at 2857
    Illegal PstI site found at 3442
    Illegal PstI site found at 4400
    Illegal PstI site found at 4628
    Illegal PstI site found at 6138
    Illegal PstI site found at 7152
    Illegal NgoMIV site found at 2701
    Illegal NgoMIV site found at 4090
    Illegal NgoMIV site found at 4264
    Illegal NgoMIV site found at 4414
    Illegal NgoMIV site found at 7588
    Illegal AgeI site found at 4715
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

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_K3726038” L0_MCGII which contains the other half part of the MCG cycle.

The other half of the MCG cycle is going to be expressed in the part PKMCGII_Lv1 which has two variants “BBa_K3726053” L1_PKMCGII-A and “BBa_K3726054” L1_PKMCGII-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_K3726110” 5CON1(H)_nNS4-up (PCC 11801) and “BBa_K3726111” 3CON5(H)_nNS4-down (PCC 11801) for the double homologous recombination within the genome of PCC 11801.

Likewise, an ampicillin 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_Ppc “BBa_K3726091”.

Source

This construct has been made by golden gate reaction.

References

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