Difference between revisions of "Part:BBa K3970025"

Revision as of 14:53, 14 October 2021

pGAP-PcyA-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

catgccgagcaaatgcctgcaaatcgctccccatttctctagtcattatcaatactgccatttcaaagaatacgtaaataattaatagtagtgattttcctaactttatttagtcaaaaaattagccttttaattctgctgtaacccgtacatgcccaaaatagggggcgggttacacagaatatataacatcgtaggtgtctgggtgaacagtttattcctggcatccactaaatataatggagcccgctttttaagctggcatccagaaaaaaaaagaatcccagcaccaaaatattgttttcttcaccaaccatcagttcataggtccattctcttagcgcaactacagagaacaggggcacaaacaggcaaaaaacgggcacaacctcaatggagtgatgcaacctgcctggagtaaatgatgacacaaggcaattgacccacgcatgtatctatctcattttcttacaccttctattaccttctgctctctctgatttggaaaaagctgaaaaaaaaggttgaaaccagttccctgaaattattcccctacttgactaataagtatataaagacggtaggtattgattgtaattctgtaaatctatttcttaaacttcttaaattctacttttatagttagtcttttttttagttttaaaacaccaagaacttagtttcgaataaacacacataaacaaacaaaagatctgatatggatcgaattagatctcgccaccATGAAGACCCCTATCACCGAAGCCATTGCCGCTGCTGACACTCAAGGCCGTTTTTTGTCCAACACTGAATTGCAAGCTGCTGATGGTAGATTCAAGAGAGCTGTTGCTTCCATGGAAGCTGCCAGAGCTTTGACCAACAACGCCCAATCTTTGATTGATGGTGCTGCACAAGCCGTCTACCAAAAGTTCCCATACACTACCACCATGCAAGGTTCTCAATATGCTTCTACTCCAGAAGGTAAGGCTAAGTGTGCTAGAGATATCGGTTACTACTTGAGAATGGTTACATACTGTTTGGTCGCCGGTGGTACTGGTCCAATGGACGAATACTTAATTGCTGGTTTGGCTGAAATCAACTCCACTTTCGACTTGTCTCCATCTTGGTACGTTGAAGCTTTGAAGTACATCAAAGCCAACCACGGTTTATCAGGTCAAGCTGCTGTCGAAGCTAACGCTTACATTGACTACGCTATCAATGCTCTATCCTGAgcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccgg

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]

@@ Line 47: / Line 47: @@
 <b>1-4: ADH1(terminator)</b>
 <br><br><br><br><br><br><br><br><br><br>
+==Sequence==
+<p style="word-wrap:break-word;">catgccgagcaaatgcctgcaaatcgctccccatttctctagtcattatcaatactgccatttcaaagaatacgtaaataattaatagtagtgattttcctaactttatttagtcaaaaaattagccttttaattctgctgtaacccgtacatgcccaaaatagggggcgggttacacagaatatataacatcgtaggtgtctgggtgaacagtttattcctggcatccactaaatataatggagcccgctttttaagctggcatccagaaaaaaaaagaatcccagcaccaaaatattgttttcttcaccaaccatcagttcataggtccattctcttagcgcaactacagagaacaggggcacaaacaggcaaaaaacgggcacaacctcaatggagtgatgcaacctgcctggagtaaatgatgacacaaggcaattgacccacgcatgtatctatctcattttcttacaccttctattaccttctgctctctctgatttggaaaaagctgaaaaaaaaggttgaaaccagttccctgaaattattcccctacttgactaataagtatataaagacggtaggtattgattgtaattctgtaaatctatttcttaaacttcttaaattctacttttatagttagtcttttttttagttttaaaacaccaagaacttagtttcgaataaacacacataaacaaacaaaagatctgatatggatcgaattagatctcgccaccATGAAGACCCCTATCACCGAAGCCATTGCCGCTGCTGACACTCAAGGCCGTTTTTTGTCCAACACTGAATTGCAAGCTGCTGATGGTAGATTCAAGAGAGCTGTTGCTTCCATGGAAGCTGCCAGAGCTTTGACCAACAACGCCCAATCTTTGATTGATGGTGCTGCACAAGCCGTCTACCAAAAGTTCCCATACACTACCACCATGCAAGGTTCTCAATATGCTTCTACTCCAGAAGGTAAGGCTAAGTGTGCTAGAGATATCGGTTACTACTTGAGAATGGTTACATACTGTTTGGTCGCCGGTGGTACTGGTCCAATGGACGAATACTTAATTGCTGGTTTGGCTGAAATCAACTCCACTTTCGACTTGTCTCCATCTTGGTACGTTGAAGCTTTGAAGTACATCAAAGCCAACCACGGTTTATCAGGTCAAGCTGCTGTCGAAGCTAACGCTTACATTGACTACGCTATCAATGCTCTATCCTGAgcgaatttcttatgatttatgatttttattattaaataagttataaaaaaaataagtgtatacaaattttaaagtgactcttaggttttaaaacgaaaattcttattcttgagtaactctttcctgtaggtcaggttgctttctcaggtatagcatgaggtcgctcttattgaccacacctctaccgg</p>
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 <span class='h3bb'>Sequence and Features</span>