| 质粒名称: | mAmetrine-N1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 54505 |
| 存储实验室: | Robert Campbell, Michael Davidson |
|
载体骨架基本信息 |
|
| 载体名称: | mAmetrine-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4750 bp |
| 完整质粒大小: | 4732 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | 无 |
| 标签/融合蛋白: | mAmetrine (C terminal on backbone) |
|
文献引用方法 |
|
| 材料和方法部分: | mAmetrine-N1 was a gift from Robert Campbell & Michael Davidson (Addgene plasmid # 54505 ; http://n2t.net/addgene:54505 ; RRID:Addgene_54505) |
| 参考文献部分: | Fluorescent protein FRET pairs for ratiometric imaging of dual biosensors. Ai HW, Hazelwood KL, Davidson MW, Campbell RE. Nat Methods. 2008 May . 5(5):401-3. 10.1038/nmeth.1207 PubMed 18425137 |
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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 mAmetrine-N1 4732 bp DNA circular SYN 25-AUG-2024
DEFINITION Localization: N1 Cloning Vector, Excitation: 406, Emission: 526.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4732)
AUTHORS Ai HW, Hazelwood KL, Davidson MW, Campbell RE
TITLE Fluorescent protein FRET pairs for ratiometric imaging of dual
biosensors.
JOURNAL Nat Methods. 2008 May . 5(5):401-3.
PUBMED 18425137
REFERENCE 2 (bases 1 to 4732)
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"; journalName: "Nat
Methods. 2008 May . 5(5):401-3."
FEATURES Location/Qualifiers
source 1..4732
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 531..611
/label=MCS
/note="multiple cloning site"
regulatory 613..622
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 619..1338
/codon_start=1
/product="yellow fluorescent protein with a large Stokes
shift, excitable with violet light (Ai et al., 2008)"
/label=mAmetrine
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTSGKLPVPWPTLVTTLSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GSYRTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNMNVWDAYITADKQKNGIK
ANFKIEHNVEDGGVQLADAYQQNTPIGDGSVLLPDNHYLSFQSKLFKDPNEQRDHMVLL
EFVTAAGITPGMDELYK"
primer_bind complement(664..685)
/label=EGFP-N
/note="EGFP, reverse primer"
polyA_signal 1460..1581
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1497..1516)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1551..1570
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1588..2043)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1725..1746)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1937..1956
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2070..2174
/gene="bla"
/label=AmpR promoter
promoter 2176..2533
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2176..2367
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2199..2219)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2384..2519
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2568..3362
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 2568..3362
/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(2622..2641)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3232..3251
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3550..3569)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3594..3641
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 3970..4558
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4459..4478
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
541 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
601 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
661 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
721 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcacctc cggcaagctg
781 cccgtgccct ggcccaccct cgtgaccacc ctgtcttacg gcgtgcagtg cttcgcccgc
841 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
901 caggagcgca ccatctcctt caaggacgac ggcagctaca ggacccgcgc cgaggtgaag
961 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
1021 ggcaacatcc tggggcacaa gctggagtac aacatgaacg tgtgggacgc gtatatcacg
1081 gccgacaagc agaagaacgg catcaaagcg aacttcaaga tcgagcacaa cgtcgaggac
1141 ggcggcgtgc agctcgccga cgcgtaccag cagaacaccc ccatcggcga cggctccgtg
1201 ctgctgcctg acaaccacta cctgagcttc cagagcaagc tgttcaaaga ccccaacgag
1261 cagcgcgatc acatggtcct gctggagttc gttaccgccg ccgggatcac tcccggcatg
1321 gacgagctgt acaagtaagc ggccgcgact ctagatcata atcagccata ccacatttgt
1381 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
1441 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
1501 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
1561 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
1621 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
1681 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
1741 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
1801 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
1861 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
1921 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
1981 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
2041 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
2101 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
2161 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
2221 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
2281 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
2341 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
2401 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
2461 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
2521 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
2581 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
2641 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
2701 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
2761 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
2821 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
2881 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
2941 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
3001 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
3061 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
3121 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
3181 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
3241 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
3301 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
3361 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
3421 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
3481 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
3541 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
3601 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
3661 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
3721 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
3781 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
3841 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
3901 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
3961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
4021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
4081 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
4441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
4501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
4561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
4621 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
4681 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cg
//
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