| 质粒名称: | pLAMP1-mCherry |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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 |
买家导航
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
//
