pAmCyan

保存条件 ：-20℃

保质期 ：6个月

英文名 ：pAmCyan

库存 ：大量

供应商 ：钰博生物

规格 ：1ug/5ug

pAmCyan

载体基本信息

出品公司:	Ybscience
载体名称:	pAmCyan
质粒类型:	大肠杆菌表达载体；cDNA克隆载体；荧光报告载体
高拷贝/低拷贝:	高拷贝
克隆方法:	限制性内切酶；多克隆位点
启动子:	lac
载体大小:	3325bp
5' 测序引物及序列:	--
3' 测序引物及序列:	--
载体标签:	--
载体抗性:	氨苄青霉素（Ampicillin）
克隆菌株:	DH5alpha等
表达菌株:	大肠杆菌
备注:	pAmCyan载体表达青色荧光蛋白AmCyan，也可以表达N端AmCyan融合蛋白； pAmCyan载体也可作为AmCyan蛋白cDNA的克隆载体， 在AmCyan基因的5‘端和3’端各有一个MCS区，便于将AmCyan基因片段转移到其它载体上； AmCyan1具有高度可溶性，高亮度，同时与其它荧光蛋白颜色区别大。
稳定性:	稳表达
组成型/诱导型:	诱导型（IPTG诱导)
病毒/非病毒:	非病毒

 

pAmCyan载体质粒图谱和多克隆位点信息

载体简介

 

pAmCyan载体描述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载体序列

LOCUS       pAmCyan	3325 bp 	DNA	SYNDEFINITION  pAmCyanACCESSION   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//