Academic Integrity: tutoring, explanations, and feedback — we don’t complete graded work or submit on a student’s behalf.

Using the webpate http://bioinfoc.ch/sawicgi/sawi.cgi?remap , please determine t

ID: 74010 • Letter: U

Question

Using the webpate http://bioinfoc.ch/sawicgi/sawi.cgi?remap , please determine the location of the two internal EcoRV sites in the sequence and explain what single nucleotide changes can remove the two EcoRV sites without changing the amino acid sequence.

       1 ATGTGCCACC AGCAGCTGGT GATCAGCTGG TTCAGCCTGG TGTTCCTGGC

      51 CAGCCCCCTG GTGGCCATCT GGGAGCTGAA GAAGGACGTG TACGTGGTGG

     101 AGCTGGACTG GTACCCCGAC GCCCCCGGCG AGATGGTGGT GCTGACCTGC

     151 GACACCCCCG AGGAGGACGG CATCACCTGG ACCCTGGACC AGAGCAGCGA

   201 GGTGCTGGGC AGCGGCAAGA CCCTGACCAT CCAGGTGAAG GAGTTCGGCG

     251 ACGCCGGCCA GTACACCTGC CACAAGGGCG GCGAGGTGCT GAGCCACAGC

     301 CTGCTGCTGC TGCACAAGAA GGAGGACGGC ATCTGGAGCA CCGATATCCT

     351 GAAGGACCAG AAGGAGCCCA AGAACAAGAC CTTCCTGAGG TGCGAGGCCA

     401 AGAACTACAG CGGCAGGTTC ACCTGCTGGT GGCTGACCAC CATCAGCACC

     451 GACCTGACCT TCAGCGTGAA GAGCAGCAGG GGCAGCAGCG ACCCCCAGGG

     501 CGTGACCTGC GGCGCCGCCA CCCTGAGCGC CGAGAGGGTG AGGGGCGACA

     551 ACAAGGAGTA CGAGTACAGC GTGGAGTGCC AGGAGGACAG CGCCTGCCCC

     601 GCCGCCGAGG AGAGCCTGCC CATCGAGGTG ATGGTGGACG CCGTGCACAA

     651 GCTGAAGTAC GAGAACTACA CCAGCAGCTT CTTCATCAGG GATATCATCA

     701 AGCCCGACCC CCCCAAGAAC CTGCAGCTGA AGCCCCTGAA GAACAGCAGG

     751 CAGGTGGAGG TGAGCTGGGA GTACCCCGAC ACCTGGAGCA CCCCCCACAG

     801 CTACTTCAGC CTGACCTTCT GCGTGCAGGT GCAGGGCAAG AGCAAGAGGG

     851 AGAAGAAGGA CAGGGTGTTC ACCGACAAGA CCAGCGCCAC CGTGATCTGC

     901 AGGAAGAACG CCAGCATCAG CGTGAGGGCC CAGGACAGGT ACTACAGCAG

    951 CAGCTGGAGC GAGTGGGCCA GCGTGCCCTG CAGC

Explanation / Answer

sequence_1.remap

SEQUENCE_1

                                        PvuII

                                        

        1 GCTGCAGGGCACGCTGGCCCACTCGCTCCAGCTGCTGCTGTAGTACCTGTCCTGGGCCCT 60

          ----:----|----:----|----:----|----:----|----:----|----:----|

        1 CGACGTCCCGTGCGACCGGGTGAGCGAGGTCGACGACGACATCATGGACAGGACCCGGGA 60

         

       61 CACGCTGATGCTGGCGTTCTTCCTGCAGATCACGGTGGCGCTGGTCTTGTCGGTGAACAC 120

          ----:----|----:----|----:----|----:----|----:----|----:----|

       61 GTGCGACTACGACCGCAAGAAGGACGTCTAGTGCCACCGCGACCAGAACAGCCACTTGTG 120

         

      121 CCTGTCCTTCTTCTCCCTCTTGCTCTTGCCCTGCACCTGCACGCAGAAGGTCAGGCTGAA 180

          ----:----|----:----|----:----|----:----|----:----|----:----|

      121 GGACAGGAAGAAGAGGGAGAACGAGAACGGGACGTGGACGTGCGTCTTCCAGTCCGACTT 180

         

      181 GTAGCTGTGGGGGGTGCTCCAGGTGTCGGGGTACTCCCAGCTCACCTCCACCTGCCTGCT 240

          ----:----|----:----|----:----|----:----|----:----|----:----|

      181 CATCGACACCCCCCACGAGGTCCACAGCCCCATGAGGGTCGAGTGGAGGTGGACGGACGA 240

                           PvuII                            EcoRV

                                                          

      241 GTTCTTCAGGGGCTTCAGCTGCAGGTTCTTGGGGGGGTCGGGCTTGATGATATCCCTGAT 300

          ----:----|----:----|----:----|----:----|----:----|----:----|

      241 CAAGAAGTCCCCGAAGTCGACGTCCAAGAACCCCCCCAGCCCGAACTACTATAGGGACTA 300

         

      301 GAAGAAGCTGCTGGTGTAGTTCTCGTACTTCAGCTTGTGCACGGCGTCCACCATCACCTC 360

          ----:----|----:----|----:----|----:----|----:----|----:----|

      301 CTTCTTCGACGACCACATCAAGAGCATGAAGTCGAACACGTGCCGCAGGTGGTAGTGGAG 360

         

      361 GATGGGCAGGCTCTCCTCGGCGGCGGGGCAGGCGCTGTCCTCCTGGCACTCCACGCTGTA 420

          ----:----|----:----|----:----|----:----|----:----|----:----|

      361 CTACCCGTCCGAGAGGAGCCGCCGCCCCGTCCGCGACAGGAGGACCGTGAGGTGCGACAT 420

                                                            DinI

                                                           

      421 CTCGTACTCCTTGTTGTCGCCCCTCACCCTCTCGGCGCTCAGGGTGGCGGCGCCGCAGGT 480

          ----:----|----:----|----:----|----:----|----:----|----:----|

      421 GAGCATGAGGAACAACAGCGGGGAGTGGGAGAGCCGCGAGTCCCACCGCCGCGGCGTCCA 480

         

      481 CACGCCCTGGGGGTCGCTGCTGCCCCTGCTGCTCTTCACGCTGAAGGTCAGGTCGGTGCT 540

          ----:----|----:----|----:----|----:----|----:----|----:----|

      481 GTGCGGGACCCCCAGCGACGACGGGGACGACGAGAAGTGCGACTTCCAGTCCAGCCACGA 540

         

      541 GATGGTGGTCAGCCACCAGCAGGTGAACCTGCCGCTGTAGTTCTTGGCCTCGCACCTCAG 600

          ----:----|----:----|----:----|----:----|----:----|----:----|

      541 CTACCACCAGTCGGTGGTCGTCCACTTGGACGGCGACATCAAGAACCGGAGCGTGGAGTC 600

                                                EcoRV

                                               

      601 GAAGGTCTTGTTCTTGGGCTCCTTCTGGTCCTTCAGGATATCGGTGCTCCAGATGCCGTC 660

          ----:----|----:----|----:----|----:----|----:----|----:----|

      601 CTTCCAGAACAAGAACCCGAGGAAGACCAGGAAGTCCTATAGCCACGAGGTCTACGGCAG 660

         

      661 CTCCTTCTTGTGCAGCAGCAGCAGGCTGTGGCTCAGCACCTCGCCGCCCTTGTGGCAGGT 720

          ----:----|----:----|----:----|----:----|----:----|----:----|

      661 GAGGAAGAACACGTCGTCGTCGTCCGACACCGAGTCGTGGAGCGGCGGGAACACCGTCCA 720

                  NaeI

                 

      721 GTACTGGCCGGCGTCGCCGAACTCCTTCACCTGGATGGTCAGGGTCTTGCCGCTGCCCAG 780

          ----:----|----:----|----:----|----:----|----:----|----:----|

      721 CATGACCGGCCGCAGCGGCTTGAGGAAGTGGACCTACCAGTCCCAGAACGGCGACGGGTC 780

         

      781 CACCTCGCTGCTCTGGTCCAGGGTCCAGGTGATGCCGTCCTCCTCGGGGGTGTCGCAGGT 840

          ----:----|----:----|----:----|----:----|----:----|----:----|

      781 GTGGAGCGACGAGACCAGGTCCCAGGTCCACTACGGCAGGAGGAGCCCCCACAGCGTCCA 840

                                                                  BtrI

                                                                  

      841 CAGCACCACCATCTCGCCGGGGGCGTCGGGGTACCAGTCCAGCTCCACCACGTACACGTC 900

          ----:----|----:----|----:----|----:----|----:----|----:----|

      841 GTCGTGGTGGTAGAGCGGCCCCCGCAGCCCCATGGTCAGGTCGAGGTGGTGCATGTGCAG 900

                             BalI           BalI                   PvuII

                                                                

      901 CTTCTTCAGCTCCCAGATGGCCACCAGGGGGCTGGCCAGGAACACCAGGCTGAACCAGCT 960

          ----:----|----:----|----:----|----:----|----:----|----:----|

      901 GAAGAAGTCGAGGGTCTACCGGTGGTCCCCCGACCGGTCCTTGTGGTCCGACTTGGTCGA 960

                  PvuII

                 

      961 GATCACCAGCTGCTGGTGGCACAT 984

          ----:----|----:----|----

      961 CTAGTGGTCGACGACCACCGTGTA 1020

# Enzymes that cut Frequency        Isoschizomers

      BalI            2    MlsI,MluNI,MscI,Msp20I

      BtrI            1    BmgBI,AjiI

      DinI            1    EgeI,EheI,SfoI

     EcoRV            2    Eco32I

      NaeI            1    PdiI

     PvuII            4   

# Enzymes which cut less frequently than the MINCUTS criterion

# Enzymes < MINCUTS Frequency        Isoschizomers

# Enzymes which cut more frequently than the MAXCUTS criterion

# Enzymes > MAXCUTS Frequency        Isoschizomers

# Enzymes that do not cut

AhaIII* BsrBI     BssNAI    Bst1107I BstZ17I   Ecl136II Eco47III Eco53kI  

EcoICRI   HpaI      MstI*     NruI      PmaCI     PmeI      PsiI      ScaI     

SmaI      SnaBI     SspI      StuI      SwaI      ZraI     

Protein sequence for the given nucleotide sequence is

sequence_1.remap

These are the outputs which you get from Remap. EcoRV is a blunt cutter having a restriction site GATATC. GAC codes for asparatic acid whereas ATC codes for Isoleucine. Alternative codons which whch code for both these amino acids are

GAC/GAU for Asp

ATT/ATC/ATA/AUC for Ile

Hence if we remove T by U (Thymine by Uracil) then EcoRV will not be able to identify the site. But there will be no change in the amino acid sequence.