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>
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<p>
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MIEQDGLHAGSPAAWVERLFGYEWAKETNYCSDSYQFSLSYQGRLTDIVSAYLAKALETLQEESS
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RLSWLAQQGVYQSAVLDSSQRSGRDWLALGEVPTTDLLTSSLNPANQKDTYKNALRRLRTLDPTK
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CELKSSSKETLSRTRSNLEASTVKKTDLYSEHEGNTTTTLTTRISARMPKKTDLKVTSRDASTSK
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IMVYNSRTTTNNDCLSETVQYSYQTITIYPRNLNSELTGTSADRILYYYGSSQPQASRIYFYRLLDEFF
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</p>
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<br/>
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<br/>
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<h1> GFP Sequence (113 amino acids + stop codon) :</h1>
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<p>
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MGKGDELLLGLVPILVELSGQVNGHRFSVSGKGTGDATRGKLTLKLACTTGSLPVSSPGLVTTLGAGLAC
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FGGSPDHRPAHKFFKSSKPEGYVQERTASAKDSGSYKMRAEVKFEGDTLANKIELRGIDSKEDGPILGTR
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REYNYNSHNAYISADAQKNGLKSNFKRRINIEDNGVQLADHYQQRLPIGDGPVLLPDNHYLSTQSQLGAD
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GNERRSHLVLLRFVTAAGITHILLPLIG
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</p>
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<br/>
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<br/>
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<h2 align="justify"><b>
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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>
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<p>
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5' -  atgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggatatgaATGG
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GCAAAGGAGACGAACTACTGCTCGGACTCGTACCAATTCTCGTTGAGCTATCAGGGCAGGTTAACGGACATC
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GTTTCAGCGTATCTGGCAAAGGCACTGGAGACGCTACAAGAGGAAAGCTCACGCTTAAGCTGGCTTGCACAA
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CAGGGAGTCTACCAGTCAGCAGTCCTGGACTCGTCACAACGCTCGGGGCGGGACTGGCTTGCTTTGGGGGAA
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GTCCCGACCACCGACCTGCTCACAAGTTCTTTAAATCCAGCAAACCAGAAGGATACGTACAAGAACGCACTG
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CGTCGGCTAAGGACTCTGGATCCTACAAAATGCGAGCTGAAGTCAAGTTCGAAGGAGACACTCTCGCGAACA
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AGATCGAACTTAGAGGCATCGACAGTAAAGAAGACGGACCTATACTCGGAACACGAAGGGAATACAACTACA
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ACTCTCACAACGCGTATATCAGCGCGGATGCCCAAAAAAACGGACTTAAAAGTAACTTCAAGAGACGCATCA
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ACATCGAAGATAATGGTGTACAACTCGCGGACCACTACCAACAACGACTGCCTATCGGAGACGGTCCAGTAC
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TCCTACCAGACAATCACTATCTATCCACGCAATCTCAACTCGGAGCTGACGGGAACGAGCGCAGATCGCATC
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TTGTACTACTACGGTTCGTCACAGCCGCAGGCATCACGCATATACTTCTACCGCTTATTGGATGAAttcttctaa - 3'
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</p>
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<br/>
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<br/>
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<h1> GFP DNA Sequence + BsaI sites :</h1>
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<p>
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5’ - ATGGGCAAAGGAGACGAACTACTGCTCGGACTCGTACCAATTCTCGTTGAGCTATCAGGGCAGGTTAACGGACATCGT
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TTCAGCGTATCTGGCAAAGGCACTGGAGACGCTACAAGAGGAAAGCTCACGCTTAAGCTGGCTTGCACAACAGGGAGTCTACCA
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GTCAGCAGTCCTGGACTCGTCACAACGCTCGGGGCGGGACTGGCTTGCTTTGGGGGAAGTCCCGACCACCGACCTGCTCACAAG
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TTCTTTAAATCCAGCAAACCAGAAGGATACGTACAAGAACGCACTGCGTCGGCTAAGGACTCTGGATCCTACAAAATGCGAGCT
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GAAGTCAAGTTCGAAGGAGACACTCTCGCGAACAAGATCGAACTTAGAGGCATCGACAGTAAAGAAGACGGACCTATACTCGGA
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ACACGAAGGGAATACAACTACAACTCTCACAACGCGTATATCAGCGCGGATGCCCAAAAAAACGGACTTAAAAGTAACTTCAAG
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AGACGCATCAACATCGAAGATAATGGTGTACAACTCGCGGACCACTACCAACAACGACTGCCTATCGGAGACGGTCCAGTACTC
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CTACCAGACAATCACTATCTATCCACGCAATCTCAACTCGGAGCTGACGGGAACGAGCGCAGATCGCATCTTGTACTACTACGG
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TTCGTCACAGCCGCAGGCATCACGCATATACTTCTACCGCTTATTGGATGA - 3’
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</p>
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<br/>
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<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:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 784
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 784
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 784
    Illegal BamHI site found at 378
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 784
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 784
  • 1000
    COMPATIBLE WITH RFC[1000]