Amino acid sequence of hGrnA for subcloning.

Identify the amino acid sequence of hGrnA for subcloning.
Design the DNA sequence encoding hGrnA optimized for Escherichia coli expression (use any K12 E. coli strain) and amenable for cloning between HindIII and XhoI restriction sites.Make sure that the back-translated optimized DNA sequence of hGrnA does not contain restriction sites HindIII and XhoI.Use genomes.urv.es/OPTIMIZER as shown during the class (codon usage HEG, method “guided random”).
Design two primers (one forward and one reverse) with melting temperatures Tm equal to approximately 60-62°C for subcloning the synthetic gene into the pET-
Highlighted with yellow amino acid sequences of all six repeats (5 points)
The amino acid sequence of hGrnA to be subcloned –use one-letter amino acid code (5 points).
The DNA sequence, encoding hGrnA, optimized for K12 E. coli expression (10 points).
Nucleotide sequences of two primers, forward and reverse, to generate a PCR product to subclone hGrnA (underline the HindIII and XhoI restriction sites) and their melting temperatures Tm. Show how you calculated the Tm. (30 points).
The resulting recombinant DNA nucleotide sequence of the construct encoding the entire amino acid sequence starting from the initiator methionine and ending with the stop codon. Underline the HindIII and XhoI restriction sites.Use the “Courier new” fonts(bold/italic/underlined) and/or different backgrounds to mark/highlight the codons for the initiator methionine, stop codon, hGrnA sequence, solubility arginine, the His-tag, and the enterokinase cleavage site (30 points).