Part:BBa_K3970024

pGAP-CpcA-tADH1

Usage and Biology

CpcA gene encodes α-phycocyanin, a light-harvesting photosynthetic bile pigment-protein from the phycobiliprotein complex (phycobilisome, PBS). Phycocyanin is the major phycobiliprotein in the PBS rod.

Construction

We use primer:XbaI-GAP-up and primer:GAP-cpcA-dn to get the fragment of GAP.

We use primer:cpcA-up and primer:cpcA-ADH1-dn to get the fragment of cpcA.

We use primer:ADH1-up and primer:ADH1-XbaI-dn to get the fragment of ADH1.

Primers we use to construct cpcA expression cassette:

XbaI-GAP-up (48-mer): tcgctccccatttctctagtcattatcaatactgccatttcaaagaat

GAP-cpcA-dn (40-mer): ttcggtgataggggtcttcatggtggcgagatctaattcg

cpcA-up (23-mer): atgaagacccctatcaccgaagc

cpcA-ADH1-dn (41-mer): tcataaatcataagaaattcgctcaggatagagcattgata

ADH1-up (30-mer): gcgaatttcttatgatttatgatttttatt

ADH1-XbaI-dn (33-mer): Cgaactctgcagtcttatatgccggtagaggt

Experiment results

the experimental pictures shows how we recycle the fragment of ADH1 terminator. ( note that how we recycle the GAP promoter and the gene cpcaA has been shown on the last parts.)

Fig. The fragments of the constructed plasmid were recovered

M: Marker

1-4: ADH1(terminator)

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]