| 出品公司: | Clontech |
|---|---|
| 載體名稱: | pAmCyan |
| 質粒類型: | 大腸桿菌表達載體；cDNA克隆載體；熒光報告載體 |
| 高拷貝/低拷貝: | 高拷貝 |
| 克隆方法: | 限制性內切酶；多克隆位點 |
| 啟動子: | lac |
| 載體大小: | 3325bp |
| 5' 測序引物及序列: | -- |
| 3' 測序引物及序列: | -- |
| 載體標簽: | -- |
| 載體抗性: | 氨芐青霉素（Ampicillin） |
| 克隆菌株: | DH5alpha等 |
| 表達菌株: | 大腸桿菌 |
| 備注: |
pAmCyan載體表達青色熒光蛋白AmCyan，也可以表達N端AmCyan融合蛋白； pAmCyan載體也可作為AmCyan蛋白cDNA的克隆載體， 在AmCyan基因的5‘端和3’端各有一個MCS區，便于將AmCyan基因片段轉移到其它載體上； AmCyan1具有高度可溶性，高亮度，同時與其它熒光蛋白顏色區別大。 |
| 產品目錄號: | 632440 |
| 穩定性: | 穩表達 |
| 組成型/誘導型: | 誘導型（IPTG誘導) |
| 病毒/非病毒: | 非病毒 |
買家導航
pAmCyan載體質粒基本信息
pAmCyan質粒圖譜載體圖譜和pAmCyan載體序列質粒序列多克隆位點信息
pAmCyan質粒載體簡介
pAmCyan載體描述
pAmCyan is a pUC19-derived prokaryotic expression vector, which encodes a variant of wildtype Anemonia majano cyan fluorescent protein (AmCyan; 1) that has been engineered for brighter fluorescence. Two amino acid substitutions (Asn-34 to Ser; Lys-68 to Met) have been made to enhance the emission characteristics of AmCyan (excitation maximum = 458 nm;emission maximum = 489 nm).
The AmCyan gene was inserted in frame with the lacZ initiation codon from pUC19 so that AmCyan is expressed from the lac promoter (Plac) in E. coli. The AmCyan coding sequence is flanked by distinct multiple cloning sites (MCS) at the 5' and 3' ends so that the gene can be readily excised from pAmCyan and subcloned into other expression vectors. An upstream sequence—located just 5' to the AmCyan 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 pAmCyan provides a high-copy-number origin of replication (pUC ori) and an ampicillin resistance gene (Ampr) for propagation and selection in E. coli.
pAmCyan Vector serves as a convenient source of AmCyan cDNA. The flanking MCS regions make it possible to excise the AmCyan coding sequence and insert it into other expression vectors. Alternatively, the AmCyan coding sequence can be amplified by PCR.
Propagation in E. coli
Recommended host strain: JM109
Selectable marker: plasmid confers resistance to ampicillin (50 μg/ml) to E. coli hosts
E. coli replication origin: pUC
Copy number: ~500
Plasmid incompatibility group: pMB1/Col E1
pAmCyan質粒序列載體序列
LOCUS pAmCyan 3325 bp DNA SYN
DEFINITION pAmCyan
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..3325
/organism="pAmCyan"
/mol_type="other DNA"
promoter 143..172
/label="lac_promoter"
misc_feature 186..208
/label="M13_pUC_rev_primer"
promoter 207..225
/label="M13_reverse_primer"
CDS 217..978
/label="ORF frame 1"
/translation="MTMITPSLHACRSTLEDPRVPVATMALSNKFIGDDMKMTYHMDG
CVNGHYFTVKGEGSGKPYEGTQTSTFKVTMANGGPLAFSFDILSTVFMYGNRCFTAYP
TSMPDYFKQAFPDGMSYERTFTYEDGGVATASWEISLKGNCFEHKSTFHGVNFPADGP
VMAKMTTGWDPSFEKMTVCDGILKGDVTAFLMLQGGGNYRCQFHTSYKTKKPVTMPPN
HAVEHRIARTDLDKGGNSVQLTEHAVAHITSVVPF*"
gene 292..975
/label="AmCyan1"
/gene="AmCyan1"
/translation="MTMITPSLHACRSTLEDPRVPVATMALSNKFIGDDMKMTYHMDG
CVNGHYFTVKGEGSGKPYEGTQTSTFKVTMANGGPLAFSFDILSTVFMYGNRCFTAYP
TSMPDYFKQAFPDGMSYERTFTYEDGGVATASWEISLKGNCFEHKSTFHGVNFPADGP
VMAKMTTGWDPSFEKMTVCDGILKGDVTAFLMLQGGGNYRCQFHTSYKTKKPVTMPPN
HAVEHRIARTDLDKGGNSVQLTEHAVAHITSVVPF*"
misc_feature complement(1115..1137)
/label="pGEX_3_primer"
/translation="MTMITPSLHACRSTLEDPRVPVATMALSNKFIGDDMKMTYHMDG
CVNGHYFTVKGEGSGKPYEGTQTSTFKVTMANGGPLAFSFDILSTVFMYGNRCFTAYP
TSMPDYFKQAFPDGMSYERTFTYEDGGVATASWEISLKGNCFEHKSTFHGVNFPADGP
VMAKMTTGWDPSFEKMTVCDGILKGDVTAFLMLQGGGNYRCQFHTSYKTKKPVTMPPN
HAVEHRIARTDLDKGGNSVQLTEHAVAHITSVVPF*"
promoter 1455..1483
/label="AmpR_promoter"
/translation="MTMITPSLHACRSTLEDPRVPVATMALSNKFIGDDMKMTYHMDG
CVNGHYFTVKGEGSGKPYEGTQTSTFKVTMANGGPLAFSFDILSTVFMYGNRCFTAYP
TSMPDYFKQAFPDGMSYERTFTYEDGGVATASWEISLKGNCFEHKSTFHGVNFPADGP
VMAKMTTGWDPSFEKMTVCDGILKGDVTAFLMLQGGGNYRCQFHTSYKTKKPVTMPPN
HAVEHRIARTDLDKGGNSVQLTEHAVAHITSVVPF*"
gene 1525..2385
/label="Ampicillin"
/gene="Ampicillin"
/translation="MTMITPSLHACRSTLEDPRVPVATMALSNKFIGDDMKMTYHMDG
CVNGHYFTVKGEGSGKPYEGTQTSTFKVTMANGGPLAFSFDILSTVFMYGNRCFTAYP
TSMPDYFKQAFPDGMSYERTFTYEDGGVATASWEISLKGNCFEHKSTFHGVNFPADGP
VMAKMTTGWDPSFEKMTVCDGILKGDVTAFLMLQGGGNYRCQFHTSYKTKKPVTMPPN
HAVEHRIARTDLDKGGNSVQLTEHAVAHITSVVPF*"
CDS 1525..2385
/label="ORF frame 1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
rep_origin 2540..3159
/label="pBR322_origin"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
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 TCGCCACCAT GGCTCTTTCA
301 AACAAGTTTA TCGGAGATGA CATGAAAATG ACCTACCATA TGGATGGCTG TGTCAATGGG
361 CATTACTTTA CCGTCAAAGG TGAAGGCAGC GGGAAGCCAT ACGAAGGGAC GCAGACCTCG
421 ACTTTTAAAG TCACCATGGC CAACGGTGGG CCCCTTGCAT TCTCCTTTGA CATACTATCT
481 ACAGTGTTCA TGTATGGAAA TCGATGCTTT ACTGCGTATC CTACCAGTAT GCCCGACTAT
541 TTCAAACAAG CATTTCCTGA CGGAATGTCA TATGAAAGGA CTTTTACCTA TGAAGATGGA
601 GGAGTTGCTA CAGCCAGTTG GGAAATAAGC CTTAAAGGCA ACTGCTTTGA GCACAAATCC
661 ACGTTTCATG GAGTGAACTT TCCTGCTGAT GGACCTGTGA TGGCGAAGAT GACAACTGGT
721 TGGGACCCAT CTTTTGAGAA AATGACTGTC TGCGATGGAA TATTGAAGGG TGATGTCACC
781 GCGTTCCTCA TGCTGCAAGG AGGTGGCAAT TACAGATGCC AATTCCACAC TTCTTACAAG
841 ACAAAAAAAC CGGTGACGAT GCCACCAAAC CATGCGGTGG AACATCGCAT TGCGAGGACC
901 GACCTTGACA AAGGTGGCAA CAGTGTTCAG CTGACGGAGC ACGCTGTTGC ACATATAACC
961 TCTGTTGTCC CTTTCTGAAG CGGCCGCGAC TCTAGAATTC CAACTGAGCG CCGGTCGCTA
1021 CCATTACCAA CTTGTCTGGT GTCAAAAATA ATAGGCCTAC TAGTCGGCCG TACGGGCCCT
1081 TTCGTCTCGC GCGTTTCGGT GATGACGGTG AAAACCTCTG ACACATGCAG CTCCCGGAGA
1141 CGGTCACAGC TTGTCTGTAA GCGGATGCCG GGAGCAGACA AGCCCGTCAG GGCGCGTCAG
1201 CGGGTGTTGG CGGGTGTCGG GGCTGGCTTA ACTATGCGGC ATCAGAGCAG ATTGTACTGA
1261 GAGTGCACCA TATGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA TACCGCATCA
1321 GGCGGCCTTA AGGGCCTCGT GATACGCCTA TTTTTATAGG TTAATGTCAT GATAATAATG
1381 GTTTCTTAGA CGTCAGGTGG CACTTTTCGG GGAAATGTGC GCGGAACCCC TATTTGTTTA
1441 TTTTTCTAAA TACATTCAAA TATGTATCCG CTCATGAGAC AATAACCCTG ATAAATGCTT
1501 CAATAATATT GAAAAAGGAA GAGTATGAGT ATTCAACATT TCCGTGTCGC CCTTATTCCC
1561 TTTTTTGCGG CATTTTGCCT TCCTGTTTTT GCTCACCCAG AAACGCTGGT GAAAGTAAAA
1621 GATGCTGAAG ATCAGTTGGG TGCACGAGTG GGTTACATCG AACTGGATCT CAACAGCGGT
1681 AAGATCCTTG AGAGTTTTCG CCCCGAAGAA CGTTTTCCAA TGATGAGCAC TTTTAAAGTT
1741 CTGCTATGTG GCGCGGTATT ATCCCGTATT GACGCCGGGC AAGAGCAACT CGGTCGCCGC
1801 ATACACTATT CTCAGAATGA CTTGGTTGAG TACTCACCAG TCACAGAAAA GCATCTTACG
1861 GATGGCATGA CAGTAAGAGA ATTATGCAGT GCTGCCATAA CCATGAGTGA TAACACTGCG
1921 GCCAACTTAC TTCTGACAAC GATCGGAGGA CCGAAGGAGC TAACCGCTTT TTTGCACAAC
1981 ATGGGGGATC ATGTAACTCG CCTTGATCGT TGGGAACCGG AGCTGAATGA AGCCATACCA
2041 AACGACGAGC GTGACACCAC GATGCCTGTA GCAATGGCAA CAACGTTGCG CAAACTATTA
2101 ACTGGCGAAC TACTTACTCT AGCTTCCCGG CAACAATTAA TAGACTGGAT GGAGGCGGAT
2161 AAAGTTGCAG GACCACTTCT GCGCTCGGCC CTTCCGGCTG GCTGGTTTAT TGCTGATAAA
2221 TCTGGAGCCG GTGAGCGTGG GTCTCGCGGT ATCATTGCAG CACTGGGGCC AGATGGTAAG
2281 CCCTCCCGTA TCGTAGTTAT CTACACGACG GGGAGTCAGG CAACTATGGA TGAACGAAAT
2341 AGACAGATCG CTGAGATAGG TGCCTCACTG ATTAAGCATT GGTAACTGTC AGACCAAGTT
2401 TACTCATATA TACTTTAGAT TGATTTAAAA CTTCATTTTT AATTTAAAAG GATCTAGGTG
2461 AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC GTGAGTTTTC GTTCCACTGA
2521 GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA
2581 ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA
2641 GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT ACCAAATACT
2701 GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC ACCGCCTACA
2761 TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA GTCGTGTCTT
2821 ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG
2881 GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG
2941 CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG GTATCCGGTA
3001 AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT
3061 CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT GTGATGCTCG
3121 TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC
3181 TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC
3241 CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC CGAGCGCAGC
3301 GAGTCAGTGA GCGAGGAAGC GGAAG
//
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pAmCyan質粒載體應用舉例
pAmCyan1-N1
2015-08-01pAmCyan1-C1
2015-08-01
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