pZsYellow is a pUC19-derived prokaryotic expression vector, which encodes a variant of wild-type Zoanthus sp. yellow fluorescent protein (ZsYellow) that has been engineered for brighter fluorescence. Wild-type ZsYellow cDNA was originally cloned from a Zoanthus species of coral, native to the Indian and Pacific oceans (1). A single amino acid substitution (Met-128 to Val) has been made to enhance the emission characteristics of ZsYellow (excitation maximum = 531 nm; emission maximum = 540 nm).

The ZsYellow gene was inserted in frame with the lacZ initiation codon from pUC19 so that ZsYellow is expressed from the lac promoter (Plac) in E. coli. The ZsYellow coding sequence is flanked by distinct multiple cloning sites (MCS) at the 5' and 3' ends so that the gene can be easily excised from pZsYellow and subcloned into other expression vectors. An upstream sequence—located just 5' to the ZsYellow gene—has been converted to a Kozak consensus translation initiation site (2) to increase the translation efficiency in eukaryotic expression systems. The pUC backbone of pZsYellow provides a high-copy-number origin of replication (pUC ori) and an ampicillin resistance gene (Ampr) for propagation and selection in E. coli.

載體應(yīng)用

pZsYellow Vector serves as a convenient source of the ZsYellow cDNA. The flanking MCS regions make it possible to excise the ZsYellow coding sequence and insert it into other prokaryotic or eukaryotic expression vectors. Alternatively, the ZsYellow coding sequence can be amplified by PCR.

0001 AGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGCTGGC 
0061 ACGACAGGTT TCCCGACTGG AAAGCGGGCA GTGAGCGCAA CGCAATTAAT GTGAGTTAGC 
0121 TCACTCATTA GGCACCCCAG GCTTTACACT TTATGCTTCC GGCTCGTATG TTGTGTGGAA 
0181 TTGTGAGCGG ATAACAATTT CACACAGGAA ACAGCTATGA CCATGATTAC GCCAAGCTTG 
0241 CATGCCTGCA GGTCGACTCT AGAGGATCCC CGGGTACCGG TCGCCACCAT GGCTCATTCA 
0301 AAGCACGGTC TAAAAGAAGA AATGACAATG AAATACCACA TGGAAGGGTG CGTCAACGGA 
0361 CATAAATTTG TGATCACGGG CGAAGGCATT GGATATCCGT TCAAAGGGAA ACAGACTATT 
0421 AATCTGTGTG TGATCGAAGG GGGACCATTG CCATTTTCCG AAGACATATT GTCAGCTGGC 
0481 TTTAAGTACG GAGACAGGAT TTTCACTGAA TATCCTCAAG ACATAGTAGA CTATTTCAAG 
0541 AACTCGTGTC CTGCTGGATA TACATGGGGC AGGTCTTTTC TCTTTGAGGA TGGAGCAGTC 
0601 TGCATATGCA ATGTAGATAT AACAGTGAGT GTCAAAGAAA ACTGCATTTA TCATAAGAGC 
0661 ATATTTAATG GAGTGAATTT TCCTGCTGAT GGACCTGTGA TGAAAAAGAT GACAACTAAC 
0721 TGGGAAGCAT CCTGCGAGAA GATCATGCCA GTACCTAAGC AGGGGATACT GAAAGGGGAT 
0781 GTCTCCATGT ACCTCCTTCT GAAGGATGGT GGGCGTTACC GGTGCCAGTT CGACACAGTT 
0841 TACAAAGCAA AGTCTGTGCC AAGTAAGATG CCGGAGTGGC ACTTCATCCA GCATAAGCTC 
0901 CTCCGTGAAG ACCGCAGCGA TGCTAAGAAT CAGAAGTGGC AGCTGACAGA GCATGCTATT 
0961 GCATTCCCTT CTGCCTTGGC CTGAAGCGGC CGCGACTCTA GAATTCCAAC TGAGCGCCGG 
1021 TCGCTACCAT TACCAACTTG TCTGGTGTCA AAAATAATAG GCCTACTAGT CGGCCGTACG 
1081 GGCCCTTTCG TCTCGCGCGT TTCGGTGATG ACGGTGAAAA CCTCTGACAC ATGCAGCTCC 
1141 CGGAGACGGT CACAGCTTGT CTGTAAGCGG ATGCCGGGAG CAGACAAGCC CGTCAGGGCG 
1201 CGTCAGCGGG TGTTGGCGGG TGTCGGGGCT GGCTTAACTA TGCGGCATCA GAGCAGATTG 
1261 TACTGAGAGT GCACCATATG CGGTGTGAAA TACCGCACAG ATGCGTAAGG AGAAAATACC 
1321 GCATCAGGCG GCCTTAAGGG CCTCGTGATA CGCCTATTTT TATAGGTTAA TGTCATGATA 
1381 ATAATGGTTT CTTAGACGTC AGGTGGCACT TTTCGGGGAA ATGTGCGCGG AACCCCTATT 
1441 TGTTTATTTT TCTAAATACA TTCAAATATG TATCCGCTCA TGAGACAATA ACCCTGATAA 
1501 ATGCTTCAAT AATATTGAAA AAGGAAGAGT ATGAGTATTC AACATTTCCG TGTCGCCCTT 
1561 ATTCCCTTTT TTGCGGCATT TTGCCTTCCT GTTTTTGCTC ACCCAGAAAC GCTGGTGAAA 
1621 GTAAAAGATG CTGAAGATCA GTTGGGTGCA CGAGTGGGTT ACATCGAACT GGATCTCAAC 
1681 AGCGGTAAGA TCCTTGAGAG TTTTCGCCCC GAAGAACGTT TTCCAATGAT GAGCACTTTT 
1741 AAAGTTCTGC TATGTGGCGC GGTATTATCC CGTATTGACG CCGGGCAAGA GCAACTCGGT 
1801 CGCCGCATAC ACTATTCTCA GAATGACTTG GTTGAGTACT CACCAGTCAC AGAAAAGCAT 
1861 CTTACGGATG GCATGACAGT AAGAGAATTA TGCAGTGCTG CCATAACCAT GAGTGATAAC 
1921 ACTGCGGCCA ACTTACTTCT GACAACGATC GGAGGACCGA AGGAGCTAAC CGCTTTTTTG 
1981 CACAACATGG GGGATCATGT AACTCGCCTT GATCGTTGGG AACCGGAGCT GAATGAAGCC 
2041 ATACCAAACG ACGAGCGTGA CACCACGATG CCTGTAGCAA TGGCAACAAC GTTGCGCAAA 
2101 CTATTAACTG GCGAACTACT TACTCTAGCT TCCCGGCAAC AATTAATAGA CTGGATGGAG 
2161 GCGGATAAAG TTGCAGGACC ACTTCTGCGC TCGGCCCTTC CGGCTGGCTG GTTTATTGCT 
2221 GATAAATCTG GAGCCGGTGA GCGTGGGTCT CGCGGTATCA TTGCAGCACT GGGGCCAGAT 
2281 GGTAAGCCCT CCCGTATCGT AGTTATCTAC ACGACGGGGA GTCAGGCAAC TATGGATGAA 
2341 CGAAATAGAC AGATCGCTGA GATAGGTGCC TCACTGATTA AGCATTGGTA ACTGTCAGAC 
2401 CAAGTTTACT CATATATACT TTAGATTGAT TTAAAACTTC ATTTTTAATT TAAAAGGATC 
2461 TAGGTGAAGA TCCTTTTTGA TAATCTCATG ACCAAAATCC CTTAACGTGA GTTTTCGTTC 
2521 CACTGAGCGT CAGACCCCGT AGAAAAGATC AAAGGATCTT CTTGAGATCC TTTTTTTCTG 
2581 CGCGTAATCT GCTGCTTGCA AACAAAAAAA CCACCGCTAC CAGCGGTGGT TTGTTTGCCG 
2641 GATCAAGAGC TACCAACTCT TTTTCCGAAG GTAACTGGCT TCAGCAGAGC GCAGATACCA 
2701 AATACTGTCC TTCTAGTGTA GCCGTAGTTA GGCCACCACT TCAAGAACTC TGTAGCACCG 
2761 CCTACATACC TCGCTCTGCT AATCCTGTTA CCAGTGGCTG CTGCCAGTGG CGATAAGTCG 
2821 TGTCTTACCG GGTTGGACTC AAGACGATAG TTACCGGATA AGGCGCAGCG GTCGGGCTGA 
2881 ACGGGGGGTT CGTGCACACA GCCCAGCTTG GAGCGAACGA CCTACACCGA ACTGAGATAC 
2941 CTACAGCGTG AGCTATGAGA AAGCGCCACG CTTCCCGAAG GGAGAAAGGC GGACAGGTAT 
3001 CCGGTAAGCG GCAGGGTCGG AACAGGAGAG CGCACGAGGG AGCTTCCAGG GGGAAACGCC 
3061 TGGTATCTTT ATAGTCCTGT CGGGTTTCGC CACCTCTGAC TTGAGCGTCG ATTTTTGTGA 
3121 TGCTCGTCAG GGGGGCGGAG CCTATGGAAA AACGCCAGCA ACGCGGCCTT TTTACGGTTC 
3181 CTGGCCTTTT GCTGGCCTTT TGCTCACATG TTCTTTCCTG CGTTATCCCC TGATTCTGTG 
3241 GATAACCGTA TTACCGCCTT TGAGTGAGCT GATACCGCTC GCCGCAGCCG AACGACCGAG 
3301 CGCAGCGAGT CAGTGAGCGA GGAAGCGGAA G