How do I construct a plasmid Adding GFP using pUC19?

Following websites would be beneficial to you concerning this assignment -

https://www.addgene.org/mol-bio-reference/plasmid-backround/

www.ncbi.nim.nih.gov >NCBI>Literature>Pub Med Central(PMC)

https://en.wikipedia.org/wiki/Vector_(molecular_biology)

www.molbiol-tools.ca/molecular-biology-freeware.htm

Using the standard -35 and -10 sequences for sigma-70 recognition and an ideal consensus Shine-Delgarno site.

Sequence for Promoter: TTGATATNNNNNNNNNNNNNNNNNNTATA ATNNNNNNATG

DNA Transcription Sequence (Shine-Delgarno included):

ATG(TAAGGAGGT)NNNNNNNNATGACGTGGTCTAGGAGGCAG TTTTTGACGGGGGTGGGGGTGTTGGCGGC GGTGTCTGGGACGGCGGGGAGGGTGGTGG CG (the italized eventually translates to MTWSRRQFLTGVGVLAAVSGTAGRVVA; the polypeptide that will use the TAT system)

Spacer DNA sequences between the protein and GFP if needed (four repeats of Ala-Gly-Ser):

GCTGGCTCTGCTGGCTCTGCTGGCTCTGC TGGCTCT

Finally, GFP:

ATGAGTAAAGGAGAAGAACTTTTCACTGG AGTTGTCCCAATTCTTGTTGAATTAGATG GTGATGTTAATGGGCACAAATTTTCTGTC AGTGGAGAG
GGTGAAGGTGATGCAACATACGGAAAACT TACCCTTAAATTTATTTGCACTACTGGAA AACTACCTGTTCCATGGCCAACACTTGTC ACTACTTTCTCTTATGGTG
TTCAATGCTTTTCAAGATACCCAGATCAT ATGAAACGGCATGACTTTTTCAAGAGTGC CATGCCCGAAGGTTATGTACAGGAAAGAA CTATATTTTTCAAAGATGA
CGGGAACTACAAGACACGTGCTGAAGTCA AGTTTGAAGGTGATACCCTTGTTAATAGA ATCGAGTTAAAAGGTATTGATTTTAAAGA AGATGGAAACATTCTTGG
ACACAAATTGGAATACAACTATAACTCAC ACAATGTATACATCATGGCAGACAAACAA AAGAATGGAATCAAAGTTAACTTCAAAAT TAGACACAACATTGAAGA
TGGAAGCGTTCAACTAGCAGACCATTATC AACAAAATACTCCAATTGGCGATGGCCCT GTCCTTTTACCAGACAACCATTACCTGTC CACACAATCTGCCCTTTC
GAAAGATCCCAACGAAAAGAGAGACCACA TGGTCCTTCTTGAGTTTGTAACAGCTGCT GGGATTACACATGGCATGGATGAACTATA CAAATAG

The DNA sequences above are simply put together. I also added a couple extra TAA stops on the end to ensure translation termination.

A short description: Use same restriction enzyme (perhaps EcoR1) to cut pUC19 and vector insert.

Fuse insert strand into vector by mixture of ATP and DNA Ligase. Transform E.coli K12 using calcium chloride heat shock. Plate on ampicillin and X-gal medium to obtain successfully transformed colonies utilizing the manufactured plasmid.