pAmCyan

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	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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