质粒名称: | pLAMP1-mCherry |
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出品公司: | Addgene |
目录编号: | 45147 |
存储实验室: | Amy Palmer |
载体骨架基本信息 |
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载体名称: | pcDNA3.1 |
载体出品公司: | Invitrogen |
空载体大小: | 5500bp |
完整质粒大小: | 7500bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | Mach1 |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | LAMP1-mCherry |
物种来源: | H. sapiens (human), Synthetic |
插入大小: | 2000bp |
启动子: | CMV |
标签/融合蛋白: | mCherry (C terminal on insert) |
克隆方法: | Gateway Cloning |
5’测序引物 | CMV-F |
3’测序引物 | BGH Reverse |
文献引用方法 |
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材料和方法部分: | pLAMP1-mCherry was a gift from Amy Palmer (Addgene plasmid # 45147 ; http://n2t.net/addgene:45147 ; RRID:Addgene_45147) |
参考文献部分: | Imaging type-III secretion reveals dynamics and spatial segregation of Salmonella effectors. Van Engelenburg SB, Palmer AE. Nat Methods. 2010 Apr;7(4):325-30. doi: 10.1038/nmeth.1437. Epub 2010 Mar 14. 10.1038/nmeth.1437 PubMed 20228815 |
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pLAMP1-mCherry载体质粒基本信息
pLAMP1-mCherry质粒图谱载体图谱和pLAMP1-mCherry载体序列质粒序列多克隆位点信息
pLAMP1-mCherry质粒载体简介
pLAMP1-mCherry质粒序列载体序列
LOCUS pLAMP1-mCherry 7453 bp DNA circular SYN 25-AUG-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7453) AUTHORS Van Engelenburg SB, Palmer AE TITLE Imaging type-III secretion reveals dynamics and spatial segregation of Salmonella effectors. JOURNAL Nat Methods. 2010 Apr;7(4):325-30. doi: 10.1038/nmeth.1437. Epub 2010 Mar 14. PUBMED 20228815 REFERENCE 2 (bases 1 to 7453) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.biofeng.com COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth.1437"; journalName: "Nat Methods"; date: "2010-04"; volume: "7"; issue: "4"; pages: "325-30" FEATURES Location/Qualifiers source 1..7453 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 93..472 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 473..676 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 627..647 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 721..740 /label=T7 /note="T7 promoter, forward primer" promoter 721..739 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 762..786 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 788..797 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 790..2040 /codon_start=1 /product="human lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV LIAYLVGRKRSHAGYQTI" protein_bind complement(2042..2066) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2086..2796 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(2231..2249) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2499..2518 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(2902..2919) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2908..3132 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3178..3606 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3265..3284) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3475..3496 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3615..3635) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3620..3949 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3800..3935 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3862..3881 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4016..4810 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4070..4089) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4680..4699 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4984..5105 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5021..5040) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5075..5094 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5154..5170) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5154..5170) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5167..5189) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5178..5194 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5202..5232) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5247..5268 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5385..5402) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5556..6144) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5636..5655) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6315..7175) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6938..6957 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7176..7280) /gene="bla" /label=AmpR promoter primer_bind complement(7355..7374) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct 61 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata 121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 661 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat 721 taatacgact cactataggg agacccaagc tggctagatc aacaagtttg tacaaaaaag 781 caggctacca tggcggcccc cggcagcgcc cggcgacccc tgctgctgct actgctgttg 841 ctgctgctcg gcctcatgca ttgtgcgtca gcagcaatgt ttatggtgaa aaatggcaac 901 gggaccgcgt gcataatggc caacttctct gctgccttct cagtgaacta cgacaccaag 961 agtggcccta agaacatgac ctttgacctg ccatcagatg ccacagtggt gctcaaccgc 1021 agctcctgtg gaaaagagaa cacttctgac cccagtctcg tgattgcttt tggaagagga 1081 catacactca ctctcaattt cacgagaaat gcaacacgtt acagcgtcca gctcatgagt 1141 tttgtttata acttgtcaga cacacacctt ttccccaatg cgagctccaa agaaatcaag 1201 actgtggaat ctataactga catcagggca gatatagata aaaaatacag atgtgttagt 1261 ggcacccagg tccacatgaa caacgtgacc gtaacgctcc atgatgccac catccaggcg 1321 tacctttcca acagcagctt cagcagggga gagacacgct gtgaacaaga caggccttcc 1381 ccaaccacag cgccccctgc gccacccagc ccctcgccct cacccgtgcc caagagcccc 1441 tctgtggaca agtacaacgt gagcggcacc aacgggacct gcctgctggc cagcatgggg 1501 ctgcagctga acctcaccta tgagaggaag gacaacacga cggtgacaag gcttctcaac 1561 atcaacccca acaagacctc ggccagcggg agctgcggcg cccacctggt gactctggag 1621 ctgcacagcg agggcaccac cgtcctgctc ttccagttcg ggatgaatgc aagttctagc 1681 cggtttttcc tacaaggaat ccagttgaat acaattcttc ctgacgccag agaccctgcc 1741 tttaaagctg ccaacggctc cctgcgagcg ctgcaggcca cagtcggcaa ttcctacaag 1801 tgcaacgcgg aggagcacgt ccgtgtcacg aaggcgtttt cagtcaatat attcaaagtg 1861 tgggtccagg ctttcaaggt ggaaggtggc cagtttggct ctgtggagga gtgtctgctg 1921 gacgagaaca gcatgctgat ccccatcgct gtgggtggtg ccctggcggg gctggtcctc 1981 atcgtcctca tcgcctacct cgtcggcagg aagaggagtc acgcaggcta ccagactatc 2041 cacccagctt tcttgtacaa agtggttgat ctagcgttta ccggtatggt gagcaagggc 2101 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc 2161 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc 2221 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc 2281 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc 2341 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag 2401 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac 2461 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc 2521 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag 2581 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc 2641 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac 2701 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc 2761 cactccaccg gcggcatgga cgagctgtac aagtaaggta ccgagctcgg atccactagt 2821 ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc 2881 ccgttgaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 2941 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 3001 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 3061 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 3121 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 3181 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 3241 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 3301 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 3361 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 3421 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 3481 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 3541 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 3601 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 3661 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 3721 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 3781 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 3841 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 3901 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 3961 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat 4021 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 4081 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 4141 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga 4201 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 4261 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 4321 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 4381 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 4441 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 4501 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga 4561 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 4621 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 4681 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 4741 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 4801 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 4861 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4921 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4981 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 5041 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 5101 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 5161 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 5221 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 5281 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 5341 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 5401 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 5461 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 5521 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 5581 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 5641 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5701 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 5761 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 5821 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5881 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5941 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 6001 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 6061 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 6121 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 6181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 6241 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 6301 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 6361 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 6421 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 6481 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 6541 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 6601 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 6661 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 6721 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 6781 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6841 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6901 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6961 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 7021 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 7081 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 7141 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 7201 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 7261 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg 7321 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt 7381 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa 7441 tttaagctac aac //
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pLAMP1-mCherry质粒载体应用举例
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