Question

31. You decide you would like to tag ezrin with GFP forimaging within cells, however, you would like to replace thecentral alpha helical linker with the GFP. Below are thesequences you will need. To accomplish this you must PCR outthe GFP gene and ligate it into the middle of the ezrin sequence. Please design the primers required for PCR (assume 15 baseshomology) and complete the final construct map. (9pts) Common start: ATG Common stop: TAA GFP atg gtg agc aag ggc gag gag ctg ttc acc ggg gtg gtg cccatc ctg gtc gag ctg gac ggc gac gta aac ggc cac aag ttc agc gtg tccggc gag ………………. agt tcg tga ccg ccg ccg gga tca ctc tcg gca tgg acgagc tgt aca agt ccg gcc gga ctc aga tct cga gct taa Insulin Hind II Nde I ATG CCG AAA CCA ATC AAT GTC CGA GTT ACC ACCATG GAT GCA GAG CTG GAG TTT GCA ATC CAG CCA AAT ACA ACT GGA AAA CAGCTT TTT GAT CAG GTG GTA AAG ACT ATC GGC CTC TCC TCC AAG TGA AGG AAGGAA TCC TTA GCG ATG AGA TCT ACT GCC CCC CTG AGA CTG CCG TGC TCT TGGGGT CCT ACG CTG TGC AGG CAA GTT TGG GGA CTA CAA CAA AGA AGT GCA CAAGTC TGG GTA CCT CAG CTC TGA GCG GCT GAT CCC TCA AAG AGT GAT GGA CCAGCA CAA ACT TAC CAG GGA CCA GTG GGA GGA CCG GAT CCA GGT GTG GCA TGCGGA ACA CCG TGG GAT GCT CAA AGA TAA TGC TAT GTT GGA ATA CCT GAA GATTGC TCA GGA CCT GGA AAT GTA TGG AAT CAA CTA TTT CGA GAT AAA AAA CAAGAA AGG AAC AGA CCT TTG GCT TGG AGT TGA TGC CCT TGG ACT GAA TAT TTATGA GAA AGA TGA TAA GTT AAC CCC AAA GAT TGG CTT TCC TTG GAG TGA AATCAG GAA CAT CTC TTT CAA TGA CAA AAA GTT TGT CAT TAA ACC CAT CGA CAAGAA GGC ACC TGA CTT TGT GTT TTA TGC GCA ACG ATG GCT GCACTG CGG GCT AAG GAG GAG CTG GAG AGA CAG GCG GTG GAT CAG ATA AAG AGCCAG GAG CAG CTG GCT GCG GAG CTT GCA GAA TAC ACT GCC AAG ATT GCC AGGGCC AAA GAA GCC CAG GAT GAC CTG GTG AAG ACC AAG GAG GAG CTG CAC CTGGTG ATG ACA GCA CCC CCG CCC CCA CCA CCC CCC GTG TAC GAG CCG GTG AGCTAC CAT GTC CAG GAG AGC TTG CAG GAT GAG GGC GCA GAG CCC ACG GGC TACAGC GCG GAG CTG TCT AGT GAG GCA TCC GGG ATG TGA GGA GAA GCG CAT CACGAG GCA TTG CAA GAG AAG AAC GAG CGT GTG CAG CGG CAG CTG CTGACG CTG AGC AGC GAG CTG TCC CAG GCC CGA GAT GAG AAT AAG AGG ACC CACAAT GAC ATC ATC CAC AAC GAG AAC ATG AGG CAA GGC CGG GAC AAG TAC AAGACG CTG CGG CAG ATC CGG CAG GGC AAC ACC AAG CAG CGC ATC GAC GAG TTCGAG GCC CTG TAA