Team:Sorbonne U Paris/Composite Part

This year our team will be submitting eleven composite parts that are used in our project:
Five parts were created (BBa_K2909004 to BBa_K2909008) for the characterization of our HiBiT tags (BBa_K2909000 and BBa_K2909001).
Four parts were created (BBa_K2909009 to BBa_K2909012) for the characterization of our enzymes (BBa_K2909002 and BBa_K2909003).
Two parts (BBa_K2909013 and BBa_K2909014) were created to be used to construct our engineered strains that we designed for our project. Unfortunately these parts do not follow the original design. They were supposed to also contain one of the two FAT enzymes as these enzymes were not successfully synthesized.

All the composite parts are linear DNA fragments that are the results of the digestion of level M plasmids by the type IIS BsaI restriction enzyme. It is these linear DNA fragments that are integrated into the genomic DNA of Chlamydomonas reinhardtii during the transformation.


COMPOSITE PARTS
PART NAME PART DESCRIPTION
BBa_K2909004 ParoR_CloverGFP
BBa_K2909005 ParoR_NanoLuc-CloverGFP
BBa_K2909006 ParoR_CloverGFP-NanoLuc
BBa_K2909007 ParoR_HiBiT-CloverGFP
BBa_K2909008 ParoR_CloverGFP-HiBiT
BBa_K2909009 HiBiT-LPAAT-A_HygroR E. guineensis
BBa_K2909010 LPAAT-A-HiBiT_HygroR E. guineensis
BBa_K2909011 HiBiT-DGAT-1-2_HygroR E. guineensis
BBa_K2909012 DGAT-1-2-HiBiT_HygroR E. guineensis
BBa_K2909013 HiBiT-DGAT-1-2_HiBiT-LPAAT-A_HygroRE. guineensis
BBa_K2909014 DGAT-1-2-HiBiT_LPAAT-A-HiBiT_HygroR E. guineensis

ParoR_CloverGFP (BBa_K2909004)



This part is a reporter construct with a Clover GFP reporter gene under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Paromomycin resistance cassette called AphVIII to select the recombinant clones.
This part has been designed to be used as a control for the characterization of our HiBiT tag parts (BBa_K2909000 and BBa_K2909001).


ParoR_NanoLuc-CloverGFP (BBa_K2909005)



This part is a reporter construct with a Clover GFP reporter gene tagged with a N-terminal NanoLuc tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Paromomycin resistance cassette called AphVIII to select the recombinant clones.

This part has been designed to be used as a control for the characterization of our HiBiT tag parts (BBa_K2909000 and BBa_K2909001).


ParoR_CloverGFP-NanoLuc (BBa_K2909006)



This part is a reporter construct with a Clover GFP reporter gene tagged with a C-terminal NanoLuc tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Paromomycin resistance cassette called AphVIII to select the recombinant clones.

This part has been designed to be used as a control for the characterization of our HiBiT tag parts (BBa_K2909000 and BBa_K2909001).


ParoR_HiBiT-CloverGFP (BBa_K2909007)



This part is a reporter construct with a Clover GFP reporter gene tagged with a N-terminal HiBiT tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Paromomycin resistance cassette called AphVIII to select the recombinant clones.

This part has been designed to be used as a reporter construct to characterize our N-terminal HiBiT tag for the Chlamydomonas reinhardtii MoClo Kit (BBa_K2909000).


ParoR_CloverGFP-HiBiT (BBa_K2909008)



This part is a reporter construct with a Clover GFP reporter gene tagged with a C-terminal HiBiT tag under the control of a nitrate-inducible Chlamydomonas reinhardtii promoter called NIT1. It also contains a Paromomycin resistance cassette called AphVIII to select the recombinant clones.

This part has been designed to be used as a reporter construct to characterize our C-terminal HiBiT tag for the Chlamydomonas reinhardtii MoClo Kit (BBa_K2909001).


HiBiT-LPAAT-A_HygroR E. guineensis (BBa_K2909009)



This part is a CDS coding for the Elaeis guineensis LPAAT-A enzyme tagged with a N-terminal HiBiT tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.

This part has been designed to characterize the expression of the Elaeis guineensis LPAAT-A enzyme that we adapted for Chlamydomonas reinhardtii (BBa_K2909002) using the N-terminal HiBiT tag (BBa_K2909000).


LPAAT-A-HiBiT_HygroR E. guineensis (BBa_K2909010)



This part is a CDS coding for the Elaeis guineensis LPAAT-A enzyme tagged with a C-terminal HiBiT tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.

This part has been designed to characterize the expression of the Elaeis guineensis LPAAT-A enzyme that we adapted for Chlamydomonas reinhardtii (BBa_K2909002) using the C-terminal HiBiT tag (BBa_K2909001).


HiBiT-DGAT-1-2_HygroR E. guineensis (BBa_K2909011)



This part is a CDS coding for the Elaeis guineensis DGAT-1-2 enzyme tagged with a N-terminal HiBiT tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.

This part has been designed to characterize the expression of the Elaeis guineensis DGAT-1-2 enzyme that we adapted for Chlamydomonas reinhardtii (BBa_K2909003) using the N-terminal HiBiT tag (BBa_K2909000).


DGAT-1-2-HiBiT_HygroR E. guineensis (BBa_K2909012)



This part is a CDS coding for the Elaeis guineensis DGAT-1-2 enzyme tagged with a C-terminal HiBiT tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.

This part has been designed to characterize the expression of the Elaeis guineensis DGAT-1-2 enzyme that we adapted for Chlamydomonas reinhardtii (BBa_K2909003) using the C-terminal HiBiT tag (BBa_K2909001).


HiBiT-DGAT-1-2_HiBiT-LPAAT-A_HygroR E. guineensis (BBa_K2909013)



This part is composed of two CDS coding for the Elaeis guineensis DGAT-1-2 (BBa_K2909003) and LPAAT-A (BBa_K2909002) enzymes, both tagged with a N-terminal HiBiT tag (BBa_K2909000).
Both enzymes are under the control of a nitrate-inducible Chlamydomonas reinhardtii promoter called NIT1.
This part also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.
This part has been designed to enhance the overall production of triglycerides in Chlamydomonas reinhardtii. The N-terminal HiBiT tag allows a quick determination of the enzyme expression.


DGAT-1-2-HiBiT_LPAAT-A-HiBiT_HygroR E. guineensis (BBa_K2909014)



This part is composed of two CDS coding for the Elaeis guineensis DGAT-1-2 (BBa_K2909003) and LPAAT-A (BBa_K2909002) enzymes, both tagged with a C-terminal HiBiT tag (BBa_K2909001).
Both enzymes are under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1.
This part also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.
This part has been designed to enhance the overall production of triglycerides in Chlamydomonas reinhardtii. The C-terminal HiBiT tag allows a quick determination of the enzyme expression.