Team:UPF CRG Barcelona/Intermediate

CONSTRUCTION INTERMEDIATES

Bba_K2581000: pFadBA with weak RBS and double terminator

It is the construction intermediate used in the BBa_K2581004 part. It consists of the assembly of a double terminator which allows forward and reverse terminator (BBa_B0014), a fatty acyl-CoA responsive promoter which expresses the downstream gene in the presence of fatty acids (BBa_K817002), and a weak RBS (BBa_B0032).

BBa_K2581001: pFadBA with weak RBS and double terminator

This is the construction intermediate used in the BBa_K2581005 part. It consists of the assembly of a double terminator which allows forward and reverse terminator (BBa_B0014), a fatty acyl-CoA responsive promoter which expresses the downstream gene in the presence of fatty acids (BBa_K817002), and a weak RBS (BBa_B0033).

BBa_K2581002: pFadBA with strong RBS and double terminator

This is the construction intermediate used in the BBa_K2581006 part. It consists of the assembly of a double terminator which allows forward and reverse terminator (BBa_B0014), a fatty acyl-CoA responsive promoter which expresses the downstream gene in the presence of fatty acids (BBa_K817002), and a strong RBS (BBa_B0034).

BBa_K2581003: FadD with weak RBS

This is the construction intermediate used in the BBa_K2581009 part. It consists in the assembly of the weak RBS 32 (BBa_B0032) and the FadD gene (BBa_K2581016) obtained of the own E. coli DH5-alpha strain genome.

BBa_K2581013: pAR: Improved fatty acyl-CoA responsive promoter

The UPF_CRG_Barcelona iGEM team 2018 has create this part as an improved element from the existing fatty acid intracellular promoter pFadBA (BBa_k817002). This new promoter has two binding sites for FadR in comparison with the existing part. Therefore, we have demonstrate an enhanced function of this promoter resulting in a disminution of basility and hence, an improvement of this part. This biobrick consists in the assembly of a double terminator which allows forward and reverse terminator (BBa_B0014), and the improved promoter based on the previous pFadBA DNA sequence.

T14_pAR part

BBa_K2581017: Intermediate fatty acid biosensor with sharp response

The UPF_CRG_Barcelona iGEM team 2018 has created this part as a intracellular biosensor of fatty acid uptake consisting in a high pass filter in order to reduce the high basal activity of the promoter in another efficient way. It consists of the assembly of a double terminator which allows forward and reverse terminator (BBa_B0014), a fatty acyl-CoA responsive promoter which expresses the downstream gene in the presence of fatty acids (BBa_K817002), a strong RBS (BBa_B0034), a CI repressor from E. coli phage lambda (+ LVA) (BBa_C0051), again the double terminator (BBa_B0014), a modified lambda Prm promoter (BBa_I12036), again the strong RBS (BBa_B0034) and an engineered mutant of red fluorescent protein from Discosoma striata (BBa_E1010).

T14_pAR part

BBa_K2581018: Intermediate fatty acid biosensor with sharp response

The UPF_CRG_Barcelona iGEM team 2018 has created this part as a intracellular biosensor of fatty acid uptake consisting in a high pass filter in order to reduce the high basal activity of the promoter in another efficient way. It consists of the assembly of a double terminator which allows forward and reverse terminator (BBa_B0014), a fatty acyl-CoA responsive promoter which expresses the downstream gene in the presence of fatty acids (BBa_K817002), a weak RBS (BBa_B0032), a CI repressor from E. coli phage lambda (+LVA) (BBa_C0051), again the double terminator (BBa_B0014), a modified lambda Prm promoter (BBa_I12036), again the weak RBS (BBa_B0032), and an engineered mutant of red fluorescent protein from Discosoma striata (BBa_E1010).

T14_pAR part

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