| 质粒名称: | LAMP1-mGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 34831 |
| 存储实验室: | Esteban Dell'Angelica |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N3 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4729 bp |
| 完整质粒大小: | 6015 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin |
| 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: | LAMP1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1300 bp |
| 启动子: | CMV |
| 标签/融合蛋白: |
mGFP (C terminal on backbone)
Kozak (N terminal on insert)
GSTGSTGSTGA linker (C terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (not destroyed) |
| 3’克隆位点: | SalI (not destroyed) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: |
LAMP1-mGFP was a gift from Esteban Dell'Angelica (Addgene plasmid # 34831 ; http://n2t.net/addgene:34831 ; RRID:Addgene_34831)
|
| 参考文献部分: | Distribution and dynamics of Lamp1-containing endocytic organelles in fibroblasts deficient in BLOC-3. Falcon-Perez JM, Nazarian R, Sabatti C, Dell'Angelica EC. J Cell Sci. 2005 Nov 15;118(Pt 22):5243-55. Epub 2005 Oct 25. 10.1242/jcs.02633 PubMed 16249233 |
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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 LAMP1-mGFP 6015 bp ds-DNA circular SYN 11-MAR-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS LAMP1-mGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6015)
AUTHORS Falcon-Perez JM, Nazarian R, Sabatti C, Dell'Angelica EC
TITLE Distribution and dynamics of Lamp1-containing endocytic organelles
in fibroblasts deficient in BLOC-3.
JOURNAL J Cell Sci. 2005 Nov 15;118(Pt 22):5243-55. Epub 2005 Oct 25.
PUBMED 16249233
REFERENCE 2 (bases 1 to 6015)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 11, 2024 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6015
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 54..357
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 358..561
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 512..532
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 634..1884
/codon_start=1
/product="human lysosome-associated membrane glycoprotein
1"
/label=LAMP1
/translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
LIAYLVGRKRSHAGYQTI"
regulatory 1948..1957
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1954..2673
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
EFVTAAGITLGMDELYK"
primer_bind complement(1999..2020)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2260..2279)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
polyA_signal 2796..2917
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2833..2852)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2887..2906
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2924..3379)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3061..3082)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3273..3292
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3406..3510
/gene="bla"
/label=AmpR promoter
promoter 3512..3869
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3535..3555)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3720..3855
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3782..3801
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3904..4698
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3958..3977)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4568..4587
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4886..4905)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4930..4977
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5306..5894
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5795..5814
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
601 cagatctcga gctcaagctt cgaattcagg gacatggcgg cccccggcag cgcccggcga
661 cccctgctgc tgctactgct gttgctgctg ctcggcctca tgcattgtgc gtcagcagca
721 atgtttatgg tgaaaaatgg caacgggacc gcgtgcataa tggccaactt ctctgctgcc
781 ttctcagtga actacgacac caagagtggc cctaagaaca tgacctttga cctgccatca
841 gatgccacag tggtgctcaa ccgcagctcc tgtggaaaag agaacacttc tgaccccagt
901 ctcgtgattg cttttggaag aggacataca ctcactctca atttcacgag aaatgcaaca
961 cgttacagcg tccagctcat gagttttgtt tataacttgt cagacacaca ccttttcccc
1021 aatgcgagct ccaaagaaat caagactgtg gaatctataa ctgacatcag ggcagatata
1081 gataaaaaat acagatgtgt tagtggcacc caggtccaca tgaacaacgt gaccgtaacg
1141 ctccatgatg ccaccatcca ggcgtacctt tccaacagca gcttcagcag gggagagaca
1201 cgctgtgaac aagacaggcc ttccccaacc acagcgcccc ctgcgccacc cagcccctcg
1261 ccctcacccg tgcccaagag cccctctgtg gacaagtaca acgtgagcgg caccaacggg
1321 acctgcctgc tggccagcat ggggctgcag ctgaacctca cctatgagag gaaggacaac
1381 acgacggtga caaggcttct caacatcaac cccaacaaga cctcggccag cgggagctgc
1441 ggcgcccacc tggtgactct ggagctgcac agcgagggca ccaccgtcct gctcttccag
1501 ttcgggatga atgcaagttc tagccggttt ttcctacaag gaatccagtt gaatacaatt
1561 cttcctgacg ccagagaccc cgcctttaaa gctgccaacg gctccctgcg agcgctgcag
1621 gccacagtcg gcaattccta caagtgcaac gcggaggagc acgtccgtgt cacgaaggcg
1681 ttttcagtca atatattcaa agtgtgggtc caggctttca aggtggaagg tggccagttt
1741 ggctctgtgg aggagtgtct gctggacgag aacagcatgc tgatccccat cgctgtgggt
1801 ggtgccctgg cggggctggt cctcatcgtc ctcatcgcct acctcgtcgg caggaagagg
1861 agtcacgcag gctaccagac tatcggctcc accggctcca caggctccac cggcgcggtc
1921 gacggtaccg cgggcccggg atccatcgcc accatggtga gcaagggcga ggagctgttc
1981 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
2041 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
2101 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
2161 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
2221 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
2281 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
2341 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
2401 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
2461 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
2521 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
2581 aaagacccca acgagaagcg cgatcacatg gtccttaagg agttcgtgac cgccgccggg
2641 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
2701 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
2761 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
2821 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
2881 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
2941 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
3001 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
3061 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
3121 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
3181 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
3241 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
3301 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
3361 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
3421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
3481 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
3541 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
3601 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
3661 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
3721 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
3781 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
3841 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
3901 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
3961 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
4021 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
4081 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
4141 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
4201 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
4261 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
4321 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
4381 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
4441 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
4501 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
4561 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
4621 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
4681 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
4741 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
4801 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
4861 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
4921 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
4981 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
5041 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
5101 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
5161 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
5221 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
5281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
5341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
5401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
5461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
5521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
5581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
5641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
5701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
5761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
5821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
5881 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
5941 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
6001 ccatgcatta gttat
//
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