Difference between revisions of "Part:BBa K3427001"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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+ | <h1> Knt Sequence (113 amino acids + stop codon) :</h1> | ||
+ | <p> | ||
+ | |||
+ | MIEQDGLHAGSPAAWVERLFGYEWAKETNYCSDSYQFSLSYQGRLTDIVSAYLAKALETLQEESS | ||
+ | RLSWLAQQGVYQSAVLDSSQRSGRDWLALGEVPTTDLLTSSLNPANQKDTYKNALRRLRTLDPTK | ||
+ | CELKSSSKETLSRTRSNLEASTVKKTDLYSEHEGNTTTTLTTRISARMPKKTDLKVTSRDASTSK | ||
+ | IMVYNSRTTTNNDCLSETVQYSYQTITIYPRNLNSELTGTSADRILYYYGSSQPQASRIYFYRLLDEFF | ||
+ | </p> | ||
+ | |||
+ | <br/> | ||
+ | <br/> | ||
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+ | <h1> GFP Sequence (113 amino acids + stop codon) :</h1> | ||
+ | <p> | ||
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+ | MGKGDELLLGLVPILVELSGQVNGHRFSVSGKGTGDATRGKLTLKLACTTGSLPVSSPGLVTTLGAGLAC | ||
+ | FGGSPDHRPAHKFFKSSKPEGYVQERTASAKDSGSYKMRAEVKFEGDTLANKIELRGIDSKEDGPILGTR | ||
+ | REYNYNSHNAYISADAQKNGLKSNFKRRINIEDNGVQLADHYQQRLPIGDGPVLLPDNHYLSTQSQLGAD | ||
+ | GNERRSHLVLLRFVTAAGITHILLPLIG | ||
+ | |||
+ | </p> | ||
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+ | <br/> | ||
+ | <br/> | ||
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+ | <h2 align="justify"><b> | ||
+ | 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. | ||
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+ | </b></h2> | ||
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+ | <h1> Knt DNA Sequence:</h1> | ||
+ | <p> | ||
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+ | 5' - atgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggatatgaATGG | ||
+ | GCAAAGGAGACGAACTACTGCTCGGACTCGTACCAATTCTCGTTGAGCTATCAGGGCAGGTTAACGGACATC | ||
+ | GTTTCAGCGTATCTGGCAAAGGCACTGGAGACGCTACAAGAGGAAAGCTCACGCTTAAGCTGGCTTGCACAA | ||
+ | CAGGGAGTCTACCAGTCAGCAGTCCTGGACTCGTCACAACGCTCGGGGCGGGACTGGCTTGCTTTGGGGGAA | ||
+ | GTCCCGACCACCGACCTGCTCACAAGTTCTTTAAATCCAGCAAACCAGAAGGATACGTACAAGAACGCACTG | ||
+ | CGTCGGCTAAGGACTCTGGATCCTACAAAATGCGAGCTGAAGTCAAGTTCGAAGGAGACACTCTCGCGAACA | ||
+ | AGATCGAACTTAGAGGCATCGACAGTAAAGAAGACGGACCTATACTCGGAACACGAAGGGAATACAACTACA | ||
+ | ACTCTCACAACGCGTATATCAGCGCGGATGCCCAAAAAAACGGACTTAAAAGTAACTTCAAGAGACGCATCA | ||
+ | ACATCGAAGATAATGGTGTACAACTCGCGGACCACTACCAACAACGACTGCCTATCGGAGACGGTCCAGTAC | ||
+ | TCCTACCAGACAATCACTATCTATCCACGCAATCTCAACTCGGAGCTGACGGGAACGAGCGCAGATCGCATC | ||
+ | TTGTACTACTACGGTTCGTCACAGCCGCAGGCATCACGCATATACTTCTACCGCTTATTGGATGAAttcttctaa - 3' | ||
+ | </p> | ||
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+ | <br/> | ||
+ | <br/> | ||
+ | |||
+ | <h1> GFP DNA Sequence + BsaI sites :</h1> | ||
+ | <p> | ||
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+ | 5’ - ATGGGCAAAGGAGACGAACTACTGCTCGGACTCGTACCAATTCTCGTTGAGCTATCAGGGCAGGTTAACGGACATCGT | ||
+ | TTCAGCGTATCTGGCAAAGGCACTGGAGACGCTACAAGAGGAAAGCTCACGCTTAAGCTGGCTTGCACAACAGGGAGTCTACCA | ||
+ | GTCAGCAGTCCTGGACTCGTCACAACGCTCGGGGCGGGACTGGCTTGCTTTGGGGGAAGTCCCGACCACCGACCTGCTCACAAG | ||
+ | TTCTTTAAATCCAGCAAACCAGAAGGATACGTACAAGAACGCACTGCGTCGGCTAAGGACTCTGGATCCTACAAAATGCGAGCT | ||
+ | GAAGTCAAGTTCGAAGGAGACACTCTCGCGAACAAGATCGAACTTAGAGGCATCGACAGTAAAGAAGACGGACCTATACTCGGA | ||
+ | ACACGAAGGGAATACAACTACAACTCTCACAACGCGTATATCAGCGCGGATGCCCAAAAAAACGGACTTAAAAGTAACTTCAAG | ||
+ | AGACGCATCAACATCGAAGATAATGGTGTACAACTCGCGGACCACTACCAACAACGACTGCCTATCGGAGACGGTCCAGTACTC | ||
+ | CTACCAGACAATCACTATCTATCCACGCAATCTCAACTCGGAGCTGACGGGAACGAGCGCAGATCGCATCTTGTACTACTACGG | ||
+ | TTCGTCACAGCCGCAGGCATCACGCATATACTTCTACCGCTTATTGGATGA - 3’ | ||
+ | </p> | ||
+ | |||
+ | <br/> | ||
+ | <br/> | ||
+ | |||
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Revision as of 14:41, 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
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 784
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 784
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 784
Illegal BamHI site found at 378 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 784
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 784
- 1000COMPATIBLE WITH RFC[1000]