1. You have sequenced a stretch of prokaryotic DNA that is cloned into the Eco R

Question

1. You have sequenced a stretch of prokaryotic DNA that is cloned into the Eco RI site of a plasmid. This fragment appears to be a complete transcription unit. Find the following sequences in this DNA:

a) two Eco RI sites
b) a promoter
c) a transcriptional terminator
d) bonus for those with nothing better to do-can you find the ORF and its start and stop codons?

Sequence:
GAATTCCGGCGGCGGGGTTTTCATTATTATGATCGTTGACATGGGACAAGGGGCTCCTATAATAGGTGCAT
TTGTAGGGGATGTGGCCACATGATAGCGCGTCGAGGCATTCAGGACCGATATTGTCATATTGCCAGCATGC
TGCAGCGCGCCAACTTAGTTACAACTTATACGATGATTTTAAAAAAAGAATATCAAATAGCCATCCACCCG
AAAGACCACCGCAAGTATCGGTACGATCGTACCAGCACACACCACAGTCGCCAGTCTGTTACCAAATAATA
AAGTTGATAATTAATTCACATCTACCTGGGTAACCCAGGTAGATGTGAATTTTTAGAATTC

Explanation / Answer

The given sequence is: GAA TTC CGG CGG CGG GGT TTT CAT TAT TAT GAT CGT TGA CAT GGG ACA AGG GGC TCC TAT AAT AGG TGC ATT TGT AGG GGA TGT GGC CAC ATG ATA GCG CGT CGA GGC ATT CAG GAC CGA TAT TGT CAT ATT GCC AGC ATG CTG  CAG CGC GCC AAC TTA GTT ACA ACT TAT ACG ATGATT TTA AAA AAA GAA TAT CAA ATA GCC ATCCAC CCG AAA GAC CAC CGC AAG TAT CGG TAC GAT CGT ACC AGC ACA CAC CAC AGT CGC CAG TCT GTT ACC AAA TAA TAA AGT TGA TAA TTA ATT CAC ATC TAC CTG GGT AAC CCA GGT AGA TGT GAA TTT TTA GAA TTC ------------------------ The restriction sites for E.coli are in red. Promoter is in pink. Transcriptional terminator is in orange. start and stop codons are in blue.

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