Difference between revisions of "Part:BBa K3427001"
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The gene was cloned in the plasmid pBAD24-MoClo under the control of an arabinose-inducible promoter. Bacteria carrying this plasmid exhibited a green fluorescence when grown in the presence of arabinose | The gene was cloned in the plasmid pBAD24-MoClo under the control of an arabinose-inducible promoter. Bacteria carrying this plasmid exhibited a green fluorescence when grown in the presence of arabinose | ||
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Revision as of 14:42, 20 October 2020
knt-gfp entangled CDSs for aminoglycoside resistance and GFP
A gene coding for the spectinomycin resistance that we named Knt and a gene coding for Green Fluorecent Protein ewas entangled.
The gene was cloned in the plasmid pBAD24-MoClo under the control of an arabinose-inducible promoter. Bacteria carrying this plasmid exhibited a green fluorescence when grown in the presence of arabinose
Knt Sequence (113 amino acids + stop codon) :
MIEQDGLHAGSPAAWVERLFGYEWAKETNYCSDSYQFSLSYQGRLTDIVSAYLAKALETLQEESS RLSWLAQQGVYQSAVLDSSQRSGRDWLALGEVPTTDLLTSSLNPANQKDTYKNALRRLRTLDPTK CELKSSSKETLSRTRSNLEASTVKKTDLYSEHEGNTTTTLTTRISARMPKKTDLKVTSRDASTSK IMVYNSRTTTNNDCLSETVQYSYQTITIYPRNLNSELTGTSADRILYYYGSSQPQASRIYFYRLLDEFF
GFP Sequence (113 amino acids + stop codon) :
MGKGDELLLGLVPILVELSGQVNGHRFSVSGKGTGDATRGKLTLKLACTTGSLPVSSPGLVTTLGAGLAC FGGSPDHRPAHKFFKSSKPEGYVQERTASAKDSGSYKMRAEVKFEGDTLANKIELRGIDSKEDGPILGTR REYNYNSHNAYISADAQKNGLKSNFKRRINIEDNGVQLADHYQQRLPIGDGPVLLPDNHYLSTQSQLGAD GNERRSHLVLLRFVTAAGITHILLPLIG
To characterize the knt-gfp entangled genes, the DNA fragment encoding the two fully overlapping CDS in different reading frames has been inserted in the plasmid pBAD24-MoClo. The entangled genes are under the control of a promoter inducible with arabinose.
Knt DNA Sequence:
5' - atgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggatatgaATGG GCAAAGGAGACGAACTACTGCTCGGACTCGTACCAATTCTCGTTGAGCTATCAGGGCAGGTTAACGGACATC GTTTCAGCGTATCTGGCAAAGGCACTGGAGACGCTACAAGAGGAAAGCTCACGCTTAAGCTGGCTTGCACAA CAGGGAGTCTACCAGTCAGCAGTCCTGGACTCGTCACAACGCTCGGGGCGGGACTGGCTTGCTTTGGGGGAA GTCCCGACCACCGACCTGCTCACAAGTTCTTTAAATCCAGCAAACCAGAAGGATACGTACAAGAACGCACTG CGTCGGCTAAGGACTCTGGATCCTACAAAATGCGAGCTGAAGTCAAGTTCGAAGGAGACACTCTCGCGAACA AGATCGAACTTAGAGGCATCGACAGTAAAGAAGACGGACCTATACTCGGAACACGAAGGGAATACAACTACA ACTCTCACAACGCGTATATCAGCGCGGATGCCCAAAAAAACGGACTTAAAAGTAACTTCAAGAGACGCATCA ACATCGAAGATAATGGTGTACAACTCGCGGACCACTACCAACAACGACTGCCTATCGGAGACGGTCCAGTAC TCCTACCAGACAATCACTATCTATCCACGCAATCTCAACTCGGAGCTGACGGGAACGAGCGCAGATCGCATC TTGTACTACTACGGTTCGTCACAGCCGCAGGCATCACGCATATACTTCTACCGCTTATTGGATGAAttcttctaa - 3'
GFP DNA Sequence + BsaI sites :
5’ - ATGGGCAAAGGAGACGAACTACTGCTCGGACTCGTACCAATTCTCGTTGAGCTATCAGGGCAGGTTAACGGACATCGT TTCAGCGTATCTGGCAAAGGCACTGGAGACGCTACAAGAGGAAAGCTCACGCTTAAGCTGGCTTGCACAACAGGGAGTCTACCA GTCAGCAGTCCTGGACTCGTCACAACGCTCGGGGCGGGACTGGCTTGCTTTGGGGGAAGTCCCGACCACCGACCTGCTCACAAG TTCTTTAAATCCAGCAAACCAGAAGGATACGTACAAGAACGCACTGCGTCGGCTAAGGACTCTGGATCCTACAAAATGCGAGCT GAAGTCAAGTTCGAAGGAGACACTCTCGCGAACAAGATCGAACTTAGAGGCATCGACAGTAAAGAAGACGGACCTATACTCGGA ACACGAAGGGAATACAACTACAACTCTCACAACGCGTATATCAGCGCGGATGCCCAAAAAAACGGACTTAAAAGTAACTTCAAG AGACGCATCAACATCGAAGATAATGGTGTACAACTCGCGGACCACTACCAACAACGACTGCCTATCGGAGACGGTCCAGTACTC CTACCAGACAATCACTATCTATCCACGCAATCTCAACTCGGAGCTGACGGGAACGAGCGCAGATCGCATCTTGTACTACTACGG TTCGTCACAGCCGCAGGCATCACGCATATACTTCTACCGCTTATTGGATGA - 3’
Sequence and Features