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pGFPuv carries
the“cycle 3”variant of GFP described by Crameri et al. (1). This gene was
cloned between the two MCSs of the pUC19 derivative pPD16.43 (2). The GFPuv
gene canbe easily excised from pGFPuv. Alternatively, the GFPuv coding sequence
can be amplified by PCR. The GFPuv gene was inserted in frame with the lacZ
initiation codon from pUC19 so that a β-galactosidase-GFPuv fusionprotein is
expressed from the lac promoter in E. coli . Note, however, that if you excise
the GFPuv coding sequence using a restriction site in the 5' MCS, the resulting
fragment will encode the native (i.e.,non-fusion) GFPuv protein. The pUC
backbone of pGFPuv provides a high copy number origin of replication and
ampicillin resistance gene for propagation in E. coli.
全序列
1 AGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGCTGGC
61 ACGACAGGTT TCCCGACTGG AAAGCGGGCA GTGAGCGCAA CGCAATTAAT GTGAGTTAGC
121 TCACTCATTA GGCACCCCAG GCTTTACACT TTATGCTTCC GGCTCGTATG TTGTGTGGAA
181 TTGTGAGCGG ATAACAATTT CACACAGGAA ACAGCTATGA CCATGATTAC GCCAAGCTTG
241 CATGCCTGCA GGTCGACTCT AGAGGATCCC CGGGTACCGG TAGAAAAAAT GAGTAAAGGA
301 GAAGAACTTT TCACTGGAGT TGTCCCAATT CTTGTTGAAT TAGATGGTGA TGTTAATGGG
361 CACAAATTTT CTGTCAGTGG AGAGGGTGAA GGTGATGCAA CATACGGAAA ACTTACCCTT
421 AAATTTATTT GCACTACTGG AAAACTACCT GTTCCATGGC CAACACTTGT CACTACTTTC
481 TCTTATGGTG TTCAATGCTT TTCCCGTTAT CCGGATCATA TGAAACGGCA TGACTTTTTC
541 AAGAGTGCCA TGCCCGAAGG TTATGTACAG GAACGCACTA TATCTTTCAA AGATGACGGG
601 AACTACAAGA CGCGTGCTGA AGTCAAGTTT GAAGGTGATA CCCTTGTTAA TCGTATCGAG
661 TTAAAAGGTA TTGATTTTAA AGAAGATGGA AACATTCTCG GACACAAACT CGAGTACAAC
721 TATAACTCAC ACAATGTATA CATCACGGCA GACAAACAAA AGAATGGAAT CAAAGCTAAC
781 TTCAAAATTC GCCACAACAT TGAAGATGGA TCCGTTCAAC TAGCAGACCA TTATCAACAA
841 AATACTCCAA TTGGCGATGG CCCTGTCCTT TTACCAGACA ACCATTACCT GTCGACACAA
901 TCTGCCCTTT CGAAAGATCC CAACGAAAAG CGTGACCACA TGGTCCTTCT TGAGTTTGTA
961 ACTGCTGCTG GGATTACACA TGGCATGGAT GAGCTCTACA AATAATGAAT TCCAACTGAG
1021 CGCCGGTCGC TACCATTACC AACTTGTCTG GTGTCAAAAA TAATAGGCCT ACTAGTCGGC
1081 CGTACGGGCC CTTTCGTCTC GCGCGTTTCG GTGATGACGG TGAAAACCTC TGACACATGC
1141 AGCTCCCGGA GACGGTCACA GCTTGTCTGT AAGCGGATGC CGGGAGCAGA CAAGCCCGTC
1201 AGGGCGCGTC AGCGGGTGTT GGCGGGTGTC GGGGCTGGCT TAACTATGCG GCATCAGAGC
1261 AGATTGTACT GAGAGTGCAC CATATGCGGT GTGAAATACC GCACAGATGC GTAAGGAGAA
1321 AATACCGCAT CAGGCGGCCT TAAGGGCCTC GTGATACGCC TATTTTTATA GGTTAATGTC
1381 ATGATAATAA TGGTTTCTTA GACGTCAGGT GGCACTTTTC GGGGAAATGT GCGCGGAACC
1441 CCTATTTGTT TATTTTTCTA AATACATTCA AATATGTATC CGCTCATGAG ACAATAACCC
1501 TGATAAATGC TTCAATAATA TTGAAAAAGG AAGAGTATGA GTATTCAACA TTTCCGTGTC
1561 GCCCTTATTC CCTTTTTTGC GGCATTTTGC CTTCCTGTTT TTGCTCACCC AGAAACGCTG
1621 GTGAAAGTAA AAGATGCTGA AGATCAGTTG GGTGCACGAG TGGGTTACAT CGAACTGGAT
1681 CTCAACAGCG GTAAGATCCT TGAGAGTTTT CGCCCCGAAG AACGTTTTCC AATGATGAGC
1741 ACTTTTAAAG TTCTGCTATG TGGCGCGGTA TTATCCCGTA TTGACGCCGG GCAAGAGCAA
1801 CTCGGTCGCC GCATACACTA TTCTCAGAAT GACTTGGTTG AGTACTCACC AGTCACAGAA
1861 AAGCATCTTA CGGATGGCAT GACAGTAAGA GAATTATGCA GTGCTGCCAT AACCATGAGT
1921 GATAACACTG CGGCCAACTT ACTTCTGACA ACGATCGGAG GACCGAAGGA GCTAACCGCT
1981 TTTTTGCACA ACATGGGGGA TCATGTAACT CGCCTTGATC GTTGGGAACC GGAGCTGAAT
2041 GAAGCCATAC CAAACGACGA GCGTGACACC ACGATGCCTG TAGCAATGGC AACAACGTTG
2101 CGCAAACTAT TAACTGGCGA ACTACTTACT CTAGCTTCCC GGCAACAATT AATAGACTGG
2161 ATGGAGGCGG ATAAAGTTGC AGGACCACTT CTGCGCTCGG CCCTTCCGGC TGGCTGGTTT
2221 ATTGCTGATA AATCTGGAGC CGGTGAGCGT GGGTCTCGCG GTATCATTGC AGCACTGGGG
2281 CCAGATGGTA AGCCCTCCCG TATCGTAGTT ATCTACACGA CGGGGAGTCA GGCAACTATG
2341 GATGAACGAA ATAGACAGAT CGCTGAGATA GGTGCCTCAC TGATTAAGCA TTGGTAACTG
2401 TCAGACCAAG TTTACTCATA TATACTTTAG ATTGATTTAA AACTTCATTT TTAATTTAAA
2461 AGGATCTAGG TGAAGATCCT TTTTGATAAT CTCATGACCA AAATCCCTTA ACGTGAGTTT
2521 TCGTTCCACT GAGCGTCAGA CCCCGTAGAA AAGATCAAAG GATCTTCTTG AGATCCTTTT
2581 TTTCTGCGCG TAATCTGCTG CTTGCAAACA AAAAAACCAC CGCTACCAGC GGTGGTTTGT
2641 TTGCCGGATC AAGAGCTACC AACTCTTTTT CCGAAGGTAA CTGGCTTCAG CAGAGCGCAG
2701 ATACCAAATA CTGTCCTTCT AGTGTAGCCG TAGTTAGGCC ACCACTTCAA GAACTCTGTA
2761 GCACCGCCTA CATACCTCGC TCTGCTAATC CTGTTACCAG TGGCTGCTGC CAGTGGCGAT
2821 AAGTCGTGTC TTACCGGGTT GGACTCAAGA CGATAGTTAC CGGATAAGGC GCAGCGGTCG
2881 GGCTGAACGG GGGGTTCGTG CACACAGCCC AGCTTGGAGC GAACGACCTA CACCGAACTG
2941 AGATACCTAC AGCGTGAGCT ATGAGAAAGC GCCACGCTTC CCGAAGGGAG AAAGGCGGAC
3001 AGGTATCCGG TAAGCGGCAG GGTCGGAACA GGAGAGCGCA CGAGGGAGCT TCCAGGGGGA
3061 AACGCCTGGT ATCTTTATAG TCCTGTCGGG TTTCGCCACC TCTGACTTGA GCGTCGATTT
3121 TTGTGATGCT CGTCAGGGGG GCGGAGCCTA TGGAAAAACG CCAGCAACGC GGCCTTTTTA
3181 CGGTTCCTGG CCTTTTGCTG GCCTTTTGCT CACATGTTCT TTCCTGCGTT ATCCCCTGAT
3241 TCTGTGGATA ACCGTATTAC CGCCTTTGAG TGAGCTGATA CCGCTCGCCG CAGCCGAACG
3301 ACCGAGCGCA GCGAGTCAGT GAGCGAGGAA GCGGAAG
