For this assignment, you will need to analyse the provided 5 sequences. For each sequence, determine whether there are any mutations to the predicted sequence, and if so whether the plasmid would still be usable in downstream applications based on whether the mutation is conservative or not. Note that sequencing can be performed on either the coding or anticoding strand. Sequences which have been performed on the anti-coding strand (using the reverse primer), which you will need to reverse complement, are denoted with a “Reveverse” in the file name. Sequences from the coding strand sequenced from the forward primer are denoted with a “Forward” in the file name.

The predicted sequence is that of the lacZ gene, cloned into pET15B, which we generated for assignment 2, and is provided below, or if you want the annotated version, it can be found in the assignment answers for assignment 2.

 

 

>Predicted-pET15B-LacZ

 

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TGGTGCCGCGCGGCAGCCACATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCGACTCTAGAGGATCCCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGtcgagcatatggctgccgcgcggcac

 

 

 

 

 

 

  1. >Forward_pET15B-LacZ_1

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taGGGGAAtnngAGCGGAtaCAATTCCCTCTAGAAAtATTTTGTTtaCTTTGGTGCCGCGCGGCAGCCACATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCGACTCTAGAGGATCCCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTCGAGCATATGGCTGCCGCGCGGCACCAGGCCGCTGCTGTGATGATGATGATGATGGCTGCTGCCCATGGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAGAGGGGAaTTGtTATCCGCTCACAATTccCCTaTAgTgnnncgtnTTaa

 

 

 

  1. >Forward_pET15B-LacZ_2

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nnnntaGggGAAtnngAGCGGAtaCAATTCCCTCTAGAAAtATTTTGTTtaCTTTGTGCCGCGCGGCAGCCACATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCGACTCTAGAGGATCCCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTCGAGCATATGGCTGCCGCGCGGCACCAGGCCGCTGCTGTGATGATGATGATGATGGCTGCTGCCCATGGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAGAGGGGAaTTGtTATCCGCTCACAATTccCCTaTAgTgnnncgtnTTaa

 

 

 

 

 

 

 

 

 

 

  1. >Forward_pET15B-LacZ_3

 

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nnnntaGggGAAtnngAGCGGAtaCnnTtccccnnnn

 

 

  1. >Reverse_pET15B-LacZ_

 

 

tcaAGaAntnAcnncgnannattGAtgnnggCAGCTTCCACAGCAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGACGCCGCTTCGTTCTACCATCGACACCACCACGCTGGCACCCAGTTGATCGGCGCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACTGGAGGTGGCAACGCCAATCAGCAACGACTGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTCTGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTACTGGTTTCACATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCATACCGCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGCGACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACCGCCGCCGCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCGAGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGAGGATCGAGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAgatataccatgggcagcagccatcatcatcatcatcacagcagcggcctggtgccgcgcggcagccatatgctcgaGTGGCTGCCGCGCGGCACCAAAGtaAACAAAATaTTTCTAGAGGGAATTGtaTCCGCTcnnaTTnnnntaggnanTnn

 

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  1. >Reverse_pET15B-LacZ_5

 

 

ntAAnangnnncAcTAtAGGggAnTTGTGAGCGGATAaCAAtTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACCATGGGCAGCAGCCATCATCATCATCATCACAGCAGCGGCCTGGTGCCGCGCGGCAGCCATATGCTCGACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTGAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCgTGCAAGCTTGGCGTAATCATGGTCATGTGGCTGCCGCGCGGCACCAAAGtaAACAAAATaTTTCTAGAGGGAATTGtaTCCGCTcnnaTTCCCCta

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