| 质粒名称: | pMSP12::dsRed2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 30171 |
| 存储实验室: | Lalita Ramakrishnan |
|
载体骨架基本信息 |
|
| 载体名称: | pFPV27 |
| 载体出品公司: | Ramakrishnan et al., 2000 |
| 空载体大小: | 4981 bp |
| 完整质粒大小: | 5277 bp |
| 载体类型: | 分枝杆菌表达载体 |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Mycobacterium Strong Promoter (MSP) |
| 物种来源: | M. marinum |
| 插入大小: | 500 bp |
| 标签/融合蛋白: | dsRed2 (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamH I(未破坏) |
| 3’克隆位点: | BamH I(未破坏) |
| 5’测序引物 | pFPV27-Fwd: GAATCGGTGGTTGTGGTGAT |
| 3’测序引物 | dsRed1-N |
|
文献引用方法 |
|
| 材料和方法部分: | pMSP12::dsRed2 was a gift from Lalita Ramakrishnan (Addgene plasmid # 30171 ; http://n2t.net/addgene:30171 ; RRID:Addgene_30171) |
| 参考文献部分: | Superinfecting mycobacteria home to established tuberculous granulomas. Cosma CL, Humbert O, Ramakrishnan L. Nat Immunol. 2004 Aug . 5(8):828-35. 10.1038/ni1091 PubMed 15220915 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pMSP12::dsRed2 5277 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMSP12::dsRed2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5277)
AUTHORS Cosma CL, Humbert O, Ramakrishnan L
TITLE Superinfecting mycobacteria home to established tuberculous
granulomas.
JOURNAL Nat Immunol. 2004 Aug . 5(8):828-35.
PUBMED 15220915
REFERENCE 2 (bases 1 to 5277)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5277
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(129..148)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 140..955
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(213..232)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 1287..1875
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1776..1795
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3938..3954
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3938..3954
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(4519..4535)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4520..4536)
/label=pBluescriptKS
/note="For pBluescript vector"
CDS 4590..5267
/codon_start=1
/product="improved tetrameric variant of DsRed fluorescent
protein"
/label=DsRed2
/note="mammalian codon-optimized"
/translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
LFL"
primer_bind complement(4770..4790)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 5182..5205
/label=DsRed1-C
/note="DsRed1, forward primer"
ORIGIN
1 tctagccacc tgacgtcggg ggggggggga aagccacgtt gtgtctcaaa atctctgatg
61 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
121 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct cgaggccgcg
181 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
241 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc catgcgccag agttgtttct
301 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
361 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
421 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
481 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
541 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
601 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
661 tgttgaacaa gtctggaaag aaatgcataa tcttttgcca ttctcaccgg attcagtcgt
721 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
781 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
841 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
901 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
961 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcggctt
1021 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
1081 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
1141 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggcctgg
1201 tatgagtcag caacaccttc ttcacgaggc agacctcact agttccactg agcgtcagac
1261 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
1321 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
1381 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
1441 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
1501 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
1561 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
1621 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
1681 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
1741 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
1801 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
1861 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
1921 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
1981 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcaa cgcgtgagcc
2041 caccagctcc gtaagttcgg gtgctgtgtg gctcgtaccc gcgcattcag gcggcagggg
2101 gtctaacggg tctaaggcgg cgtgtacggc cgccacagcg gctcttagcg gcccggaaac
2161 gtcctcgaaa cgacgcatgt gttcctcctg gttggtacag gtggttgggg gtgctcggct
2221 gtcgctggtg tttcatcatc agggctcgac gggagagcgg gggagtgtgc agttgtgggg
2281 tggcccctca gcgaaatatc tgacttggag ctcgtgtcgg accatacacc ggtgattaat
2341 cgtggtttat tatcaagcgt gagccacgtc gccgacgaat ttgagcagct ctggctgccg
2401 tactggtccc tggcaagcga cgatctgctc gaggggatct accgccaaag ccgcgcgtcg
2461 gccctaggcc gccggtacat cgaggcgaac ccaacagcgc tggcaaacct gctggtcgtg
2521 gacgtagacc atccagacgc agcgctccga gcgctcagcg cccgggggtc ccatccgctg
2581 cccaacgcga tcgtgggcaa tcgcgccaac ggccacgcac acgcagtgtg ggcactcaac
2641 gcccctgttc cacgcaccga atacgcgcgg cgtaagccgc tcgcatacat ggcggcgtgc
2701 gccgaaggcc ttcggcgcgc cgtcgatggc gaccgcagtt actcaggcct catgaccaaa
2761 aaccccggcc acatcgcctg ggaaacggaa tggctccact cagatctcta cacactcagc
2821 cacatcgagg ccgagctcgg cgcgaacatg ccaccgccgc gctggcgtca gcagaccacg
2881 tacaaagcgg ctccgacgcc gctagggcgg aattgcgcac tgttcgattc cgtcaggttg
2941 tgggcctatc ttcccgccct catgcggatc tacctgccga cccggaacgt ggacggactc
3001 ggccgcgcga tctatgccga gtgccacgcg cgaaacgccg aatttccgtg caacgacgtg
3061 tgtcccggac cgctaccgga cagcgaggtc cgcgccatcg ccaacagcat ttggcgttgg
3121 atcacaacca agtcgcgcat ttgggcggac gggatcgtgg tctacgaggc cacactcagt
3181 gcgcgccatg cggccatctc gcggaagggc gcagcagcgc gcacggcggc gagcacagtt
3241 gcgcggcgcg caaagtccgc gtcagccatg gaggcattgc tatgagcgac ggctacagcg
3301 acggctacag cgacggctac aactggcagc cgactgtccg caaaaagcgg cgcgtgaccg
3361 ccgccgaagg cgctcgaatc accggactat ccgaacgcca cgtcgtccgg ctcgtggcgc
3421 aggaacgcag cgagtggttc gccgagcagg ctgcacgccg cgaacgcatc cgcgcctatc
3481 acgacgacga gggccactct tggccgcaaa cggccaaaca tttcgggctg catctggaca
3541 ccgttaagcg actcggctat cgggcgagga aagagcgtgc ggcagaacag gaagcggctc
3601 aaaaggccca caacgaagcc gacaatccac cgctgttcta acgcaattgg ggagcgggtg
3661 tcgcgggggt tccgtggggg gttccgttgc aacgggtcgg acaggtaaaa gtcctggtag
3721 acgctagttt tctggtttgg gccatgcctg tctcgttgcg tgtttcgttg cgtccgtttt
3781 gaataccagc cagacgagac ggggttctac gaatcttggt cgataccaag ccatttccgc
3841 tgaatatcgt ggagctcacc gccagaatcg gtggttgtgg tgatgtacgt ggcgaactcc
3901 gttgtagtgc ttgtggtggc atccgtggcg cggccgctct agaactagtg gatctgaccc
3961 gctccacaac cgctgagcag ggtggcagtc agagcgacca ggccgaccga tacgaggcgg
4021 attttgaaca ggttcactcg aagcctcccg ctgggttcgg agcatcgtcg atcacatgac
4081 ggcgtgctga caccgattcg actatgcaca gtagtaggta gccgccgcgg acggtagcgc
4141 aggcggcctc ggcgccgtcg tggcgtcgcg atggttggcg ccagacccgc ctgatcggcg
4201 tcccgaccag ggccgccggg gtggtgagcc tcggcttggt ccagccgtag tgagacggga
4261 tgtcttcccg gttgtcgggt ttgggtgggc ggtcacatgt tcgctggctg tgggcgtgtc
4321 gggggtcgtc gaagcggccg cgaacagggc tggacgatgc acgtcttgca cctcctgtca
4381 acccccaatg ttgagccgcg atgcccctga cctgcaccgt tggaccgcag tcgttagggg
4441 ggcgtgttag catggagtga cgaaaggggc ctaaatcaga tgatcccccg ggctgcagga
4501 attcgatatc aagcttatcg ataccgtcga cctcgagggg gggcccggta ccagatcttt
4561 aaatctagat ttaagaagga gatatacata tggcctcctc cgagaacgtc atcaccgagt
4621 tcatgcgctt caaggtgcgc atggagggca ccgtgaacgg ccacgagttc gagatcgagg
4681 gcgagggcga gggccgcccc tacgagggcc acaacaccgt gaagctgaag gtgaccaagg
4741 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac ggctccaagg
4801 tgtacgtgaa gcaccccgcc gacatccccg actacaagaa gctgtccttc cccgagggct
4861 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggcgaccgtg acccaggact
4921 cctccctgca ggacggctgc ttcatctaca aggtgaagtt catcggcgtg aacttcccct
4981 ccgacggccc cgtgatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
5041 acccccgcga cggcgtgctg aagggcgaga cccacaaggc cctgaagctg aaggacggcg
5101 gccactacct ggtggagttc aagtccatct acatggccaa gaagcccgtg cagctgcccg
5161 gctactacta cgtggacgcc aagctggaca tcacctccca caacgaggac tacaccatcg
5221 tggagcagta cgagcgcacc gagggccgcc accacctgtt cctgtagcgg ccgcgac
//
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