ptdTomato

保存条件 ：-20℃

保质期 ：6个月

英文名 ：ptdTomato

库存 ：大量

供应商 ：钰博生物

规格 ：1ug/5ug

ptdTomato

载体基本信息

出品公司:	Ybscience
载体名称:	ptdTomato
质粒类型:	原核表达载体；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:	4065  bp
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
筛选标记:	--
克隆菌株:	DH5α
宿主细胞（系）:	大肠杆菌
备注:	原核表达载体ptdTomato在IPTG诱导下表达tdTomato红色荧光蛋白；也可作为tdTomato基因的克隆载体。
稳定性:	稳表达
组成型/诱导型:	诱导型
病毒/非病毒:	非病毒

 

ptdTomato载体质粒图谱和多克隆位点信息

载体简介

 

ptdTomato载体描述ptdTomato encodes tdTomato, a member of the family of fruit fluorescent proteins (1) derived from the Discosoma sp. red fluorescent protein, DsRed (2). Because the Tomato protein has a tendency to dimerize, the vector was desighed with two copies of the Tomato coding region liked together to allow intramolecular dimerization.  As a result, each tdTomato RNA transcript encodes a tandem dimer of the Tomato protein (excitation and emission maxima equal 554nm and 581nm, respectively). In ptdTomato, the tdTomato coding sequence (2) is flanked by restriction sites (5' MCS and 3' MCS) that make it easy to excise the gene for use in other cloning applications. In E. coli, tdTomato is expressed from the lac promoter (Plac) as a fusion with several amino acids, including the first five amino acids of the LacZ protein. Note, however, that if the tdTomato coding sequence is excised using a restriction site in the 5' MCS, the protein will no longer be expressed as a fusion (as it is when expressed from the lac promoter).A Kozak consensus sequence is located immediately upstream of the tdTomato coding sequence to enhance translational efficiency in eukaryotic cells (3). In the tdTomato vector, the entire tdTomato expression cassette is supported by a pUC19 backbone, which contains a high copy-number origin of replication and an ampicillin resistance gene for propagation and selection in E. coli.The ptdTomato vector is primarily intended to serve as a source of tdTomato cDNA. The flanking MCS regions make it possible to excise the tdTomato coding sequence and insert it into other vector systems. The vector can also be used to express tdTomato in bacteria.For Western analysis, either the Living Colors DsRed Polyclonal Antibody (Cat. No. 632496) or DsRed Monoclonal Antibody (Cat. Nos. 632392 and 632393) can be used to detect the tdTomato protein.Propagation in E. Coli Recommended host strain: DH5α Selectable marker: plasmid confers resistance to ampicillin (50 μg/ml) in E. coli hosts. E. coli replication origin: pUC Copy number: high Plasmid incompatibility group: pMB1/ColE1Excitation and emission maxima of tdTomato Excitation maximum = 554 nm Emission maximum = 581 nm

 

ptdTomato载体序列

LOCUS       ptdTomato	4065 bp 	DNA	SYNDEFINITION  ptdTomatoACCESSION   KEYWORDS    SOURCE        ORGANISM  other sequences; artificial sequences; vectors.FEATURES             Location/Qualifiers     source          1..4065                     /organism="ptdTomato"                     /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..1719                     /label="ORF frame 1"                     /translation="MTMITPSLHACRSTLEDPRVPVATMVSKGEEVIKEFMRFKVRME                     GSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPA                     DIPDYKKLSFPEGFKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPV                     MQKKTMGWEASTERLYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYY                     YVDTKLDITSHNEDYTIVEQYERSEGRHHLFLGHGTGSTGSGSSGTASSEDNNMAVIK                     EFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMY                     GSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMR                     GTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMA                     KKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHLFLYGMDELYK*"     CDS             complement(259..972)                     /label="ORF frame 1"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     gene            292..1716                     /label="dTomato (variant)"                     /gene="dTomato (variant)"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     misc_feature    457..510                     /label="Q66M (mCherry)"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     misc_feature    884..907                     /label="dsRed1_C_primer"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     misc_feature    1610..1633                     /label="dsRed1_C_primer"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     misc_feature    complement(1855..1877)                     /label="pGEX_3_primer"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     promoter        2195..2223                     /label="AmpR_promoter"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     gene            2265..3125                     /label="Ampicillin"                     /gene="Ampicillin"                     /translation="MPQEQVVAALGALVLFHDGVVLVVGGDVQLGVHVVVAGQLHGLL                     GHVDGLELHQVVAAVLQLQGLVDLALQHAVAGVQALGGGLPAHGLLLHYGAVGGEVGA                     AHLHLVDQRAVLQGGVLGHGHQTAVLEVHHALPLEALGEGQLLVIGDVGGVLHVRLGA                     VHELGGQDVPGEGQGAALGHLQLGGLGALVGAALALALDLELVAVHGALHAHLEAHEL                     FDDLLALAHHGGDRYPGIL*"     CDS             2265..3125                     /label="ORF frame 3"                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA                     EIGASLIKHW*"     rep_origin      3280..3899                     /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 GGTGAGCAAG  301 GGCGAGGAGG TCATCAAAGA GTTCATGCGC TTCAAGGTGC GCATGGAGGG CTCCATGAAC  361 GGCCACGAGT TCGAGATCGA GGGCGAGGGC GAGGGCCGCC CCTACGAGGG CACCCAGACC  421 GCCAAGCTGA AGGTGACCAA GGGCGGCCCC CTGCCCTTCG CCTGGGACAT CCTGTCCCCC  481 CAGTTCATGT ACGGCTCCAA GGCGTACGTG AAGCACCCCG CCGACATCCC CGATTACAAG  541 AAGCTGTCCT TCCCCGAGGG CTTCAAGTGG GAGCGCGTGA TGAACTTCGA GGACGGCGGT  601 CTGGTGACCG TGACCCAGGA CTCCTCCCTG CAGGACGGCA CGCTGATCTA CAAGGTGAAG  661 ATGCGCGGCA CCAACTTCCC CCCCGACGGC CCCGTAATGC AGAAGAAGAC CATGGGCTGG  721 GAGGCCTCCA CCGAGCGCCT GTACCCCCGC GACGGCGTGC TGAAGGGCGA GATCCACCAG  781 GCCCTGAAGC TGAAGGACGG CGGCCACTAC CTGGTGGAGT TCAAGACCAT CTACATGGCC  841 AAGAAGCCCG TGCAACTGCC CGGCTACTAC TACGTGGACA CCAAGCTGGA CATCACCTCC  901 CACAACGAGG ACTACACCAT CGTGGAACAG TACGAGCGCT CCGAGGGCCG CCACCACCTG  961 TTCCTGGGGC ATGGCACCGG CAGCACCGGC AGCGGCAGCT CCGGCACCGC CTCCTCCGAG 1021 GACAACAACA TGGCCGTCAT CAAAGAGTTC ATGCGCTTCA AGGTGCGCAT GGAGGGCTCC 1081 ATGAACGGCC ACGAGTTCGA GATCGAGGGC GAGGGCGAGG GCCGCCCCTA CGAGGGCACC 1141 CAGACCGCCA AGCTGAAGGT GACCAAGGGC GGCCCCCTGC CCTTCGCCTG GGACATCCTG 1201 TCCCCCCAGT TCATGTACGG CTCCAAGGCG TACGTGAAGC ACCCCGCCGA CATCCCCGAT 1261 TACAAGAAGC TGTCCTTCCC CGAGGGCTTC AAGTGGGAGC GCGTGATGAA CTTCGAGGAC 1321 GGCGGTCTGG TGACCGTGAC CCAGGACTCC TCCCTGCAGG ACGGCACGCT GATCTACAAG 1381 GTGAAGATGC GCGGCACCAA CTTCCCCCCC GACGGCCCCG TAATGCAGAA GAAGACCATG 1441 GGCTGGGAGG CCTCCACCGA GCGCCTGTAC CCCCGCGACG GCGTGCTGAA GGGCGAGATC 1501 CACCAGGCCC TGAAGCTGAA GGACGGCGGC CACTACCTGG TGGAGTTCAA GACCATCTAC 1561 ATGGCCAAGA AGCCCGTGCA ACTGCCCGGC TACTACTACG TGGACACCAA GCTGGACATC 1621 ACCTCCCACA ACGAGGACTA CACCATCGTG GAACAGTACG AGCGCTCCGA GGGCCGCCAC 1681 CACCTGTTCC TGTACGGCAT GGACGAGCTG TACAAGTAGG CGGCCGCGAC TCTAGAATTC 1741 CAACTGAGCG CCGGTCGCTA CCATTACCAA CTTGTCTGGT GTCAAAAATA ATAGGCCTAC 1801 TAGTCGGCCG TACGGGCCCT TTCGTCTCGC GCGTTTCGGT GATGACGGTG AAAACCTCTG 1861 ACACATGCAG CTCCCGGAGA CGGTCACAGC TTGTCTGTAA GCGGATGCCG GGAGCAGACA 1921 AGCCCGTCAG GGCGCGTCAG CGGGTGTTGG CGGGTGTCGG GGCTGGCTTA ACTATGCGGC 1981 ATCAGAGCAG ATTGTACTGA GAGTGCACCA TATGCGGTGT GAAATACCGC ACAGATGCGT 2041 AAGGAGAAAA TACCGCATCA GGCGGCCTTA AGGGCCTCGT GATACGCCTA TTTTTATAGG 2101 TTAATGTCAT GATAATAATG GTTTCTTAGA CGTCAGGTGG CACTTTTCGG GGAAATGTGC 2161 GCGGAACCCC TATTTGTTTA TTTTTCTAAA TACATTCAAA TATGTATCCG CTCATGAGAC 2221 AATAACCCTG ATAAATGCTT CAATAATATT GAAAAAGGAA GAGTATGAGT ATTCAACATT 2281 TCCGTGTCGC CCTTATTCCC TTTTTTGCGG CATTTTGCCT TCCTGTTTTT GCTCACCCAG 2341 AAACGCTGGT GAAAGTAAAA GATGCTGAAG ATCAGTTGGG TGCACGAGTG GGTTACATCG 2401 AACTGGATCT CAACAGCGGT AAGATCCTTG AGAGTTTTCG CCCCGAAGAA CGTTTTCCAA 2461 TGATGAGCAC TTTTAAAGTT CTGCTATGTG GCGCGGTATT ATCCCGTATT GACGCCGGGC 2521 AAGAGCAACT CGGTCGCCGC ATACACTATT CTCAGAATGA CTTGGTTGAG TACTCACCAG 2581 TCACAGAAAA GCATCTTACG GATGGCATGA CAGTAAGAGA ATTATGCAGT GCTGCCATAA 2641 CCATGAGTGA TAACACTGCG GCCAACTTAC TTCTGACAAC GATCGGAGGA CCGAAGGAGC 2701 TAACCGCTTT TTTGCACAAC ATGGGGGATC ATGTAACTCG CCTTGATCGT TGGGAACCGG 2761 AGCTGAATGA AGCCATACCA AACGACGAGC GTGACACCAC GATGCCTGTA GCAATGGCAA 2821 CAACGTTGCG CAAACTATTA ACTGGCGAAC TACTTACTCT AGCTTCCCGG CAACAATTAA 2881 TAGACTGGAT GGAGGCGGAT AAAGTTGCAG GACCACTTCT GCGCTCGGCC CTTCCGGCTG 2941 GCTGGTTTAT TGCTGATAAA TCTGGAGCCG GTGAGCGTGG GTCTCGCGGT ATCATTGCAG 3001 CACTGGGGCC AGATGGTAAG CCCTCCCGTA TCGTAGTTAT CTACACGACG GGGAGTCAGG 3061 CAACTATGGA TGAACGAAAT AGACAGATCG CTGAGATAGG TGCCTCACTG ATTAAGCATT 3121 GGTAACTGTC AGACCAAGTT TACTCATATA TACTTTAGAT TGATTTAAAA CTTCATTTTT 3181 AATTTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC 3241 GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG 3301 ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG 3361 TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA 3421 GAGCGCAGAT ACCAAATACT GTTCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA 3481 ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA 3541 GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC 3601 AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA 3661 CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA 3721 AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC 3781 CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC 3841 GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG 3901 CCTTTTTACG GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT 3961 CCCCTGATTC TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA 4021 GCCGAACGAC CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC GGAAG//