ptdTomato

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/ColE1

Excitation and emission maxima of tdTomato
 Excitation maximum = 554 nm
 Emission maximum = 581 nm

 

ptdTomato载体序列

LOCUS       ptdTomato	4065 bp 	DNA	SYN
DEFINITION  ptdTomato
ACCESSION   
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
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