Part:BBa_K3726053

Lv.1_PKMCG-A

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 P_PpsbA*-RiboJ “BBa_K3726079” 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 9525
  Illegal EcoRI site found at 1128
  Illegal SpeI site found at 10488
  Illegal PstI site found at 575
  Illegal PstI site found at 839
  Illegal PstI site found at 4411
  Illegal PstI site found at 5040
  Illegal PstI site found at 5537
  Illegal PstI site found at 6136
  Illegal PstI site found at 7074
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1128
  Illegal EcoRI site found at 9525
  Illegal NheI site found at 9177
  Illegal NheI site found at 9204
  Illegal SpeI site found at 10488
  Illegal PstI site found at 575
  Illegal PstI site found at 839
  Illegal PstI site found at 4411
  Illegal PstI site found at 5040
  Illegal PstI site found at 5537
  Illegal PstI site found at 6136
  Illegal PstI site found at 7074
  Illegal NotI site found at 9531
  Illegal NotI site found at 10495
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1128
  Illegal EcoRI site found at 9525
  Illegal BamHI site found at 505
  Illegal BamHI site found at 2950
  Illegal BamHI site found at 10070
  Illegal XhoI site found at 9401
  Illegal XhoI site found at 11519
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 9525
  Illegal suffix found in sequence at 10488
  Illegal EcoRI site found at 1128
  Illegal PstI site found at 575
  Illegal PstI site found at 839
  Illegal PstI site found at 4411
  Illegal PstI site found at 5040
  Illegal PstI site found at 5537
  Illegal PstI site found at 6136
  Illegal PstI site found at 7074
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 9525
  Illegal EcoRI site found at 1128
  Illegal XbaI site found at 9540
  Illegal SpeI site found at 10488
  Illegal PstI site found at 575
  Illegal PstI site found at 839
  Illegal PstI site found at 4411
  Illegal PstI site found at 5040
  Illegal PstI site found at 5537
  Illegal PstI site found at 6136
  Illegal PstI site found at 7074
  Illegal NgoMIV site found at 2527
  Illegal NgoMIV site found at 4636
  Illegal NgoMIV site found at 5096
  Illegal NgoMIV site found at 8992
  Illegal NgoMIV site found at 9134
  Illegal AgeI site found at 6649
  Illegal AgeI site found at 6732
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 9555
  Illegal BsaI.rc site found at 10472