| 质粒名称: | piRFP670-N1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 45457 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4012 bp |
| 完整质粒大小: | 4948 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | iRFP670 |
| 物种来源: | Rhodopseudomonas palustris |
| 插入大小: | 936 bp |
| 基因突变: | T2A, S45T, V116I, D124E, M170L, K180M, D202L, I203V, I219V, R233H, V251I, V254C, Y258F, A283I, F307L relative to RpBphP6 |
| GenBank编号 | KC991142 |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | piRFP670-N1 was a gift from Vladislav Verkhusha (Addgene plasmid # 45457 ; http://n2t.net/addgene:45457 ; RRID:Addgene_45457) |
| 参考文献部分: | Near-infrared fluorescent proteins for multicolor in vivo imaging. Shcherbakova DM, Verkhusha VV. Nat Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521. 10.1038/nmeth.2521 PubMed 23770755 |
买家导航
piRFP670N1载体基本信息
piRFP670N1载体图谱质粒图谱和多克隆位点信息
piRFP670N1载体简介
piRFP670N1载体序列
LOCUS piRFP670-N1 4948 bp ds-DNA circular SYN 12-MAR-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS piRFP670-N1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4948)
AUTHORS Shcherbakova DM, Verkhusha VV
TITLE Near-infrared fluorescent proteins for multicolor in vivo imaging.
JOURNAL Nat Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521.
PUBMED 23770755
REFERENCE 2 (bases 1 to 4948)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 12, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4948
/organism="synthetic DNA construct"
/mol_type="other DNA"
regulatory 7..16
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 13..948
/codon_start=1
/product="near-infrared fluorescent protein with an
emission peak at 670 nm, engineered from a bacterial
phytochrome (Shcherbakova and Verkhusha, 2013)"
/label=iRFP670
/note="derived from Rhodopseudomonas palustris RpBphP6"
/translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
polyA_signal 1070..1191
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1107..1126)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1161..1180
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1198..1653)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1335..1356)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1547..1566
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 1680..1784
/gene="bla"
/label=AmpR promoter
promoter 1786..2143
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(1809..1829)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 1994..2129
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2056..2075
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2178..2972
/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(2232..2251)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2842..2861
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3160..3179)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3204..3251
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 3580..4168
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4069..4088
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 4343..4646
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4647..4850
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4801..4821
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 4873..5
/label=MCS
/note="multiple cloning site"
ORIGIN
1 ccggtcgcca ccatggcgcg taaggtcgat ctcacctcct gcgatcgcga gccgatccac
61 atccccggca gcattcagcc gtgcggctgc ctgctagcct gcgacgcgca ggcggtgcgg
121 atcacgcgca ttacggaaaa tgccggcgcg ttctttggac gcgaaactcc gcgggtcggt
181 gagctactcg ccgattactt cggcgagacc gaagcccatg cgctgcgcaa cgcactggcg
241 cagtcctccg atccaaagcg accggcgctg atcttcggtt ggcgcgacgg cctgaccggc
301 cgcaccttcg acatctcact gcatcgccat gacggtacat cgatcatcga gttcgagcct
361 gcggcggccg aacaggccga caatccgctg cggctgacgc ggcagatcat cgcgcgcacc
421 aaagaactga agtcgctcga agagatggcc gcacgggtgc cgcgctatct gcaggcgatg
481 ctcggctatc accgcgtgat gttgtaccgc ttcgcggacg acggctccgg gatggtgatc
541 ggcgaggcga agcgcagcga cctcgagagc tttctcggtc agcactttcc ggcgtcgctg
601 gtcccgcagc aggcgcggct actgtacttg aagaacgcga tccgcgtggt ctcggattcg
661 cgcggcatca gcagccggat cgtgcccgag cacgacgcct ccggcgccgc gctcgatctg
721 tcgttcgcgc acctgcgcag catctcgccc tgccatctcg aatttctgcg gaacatgggc
781 gtcagcgcct cgatgtcgct gtcgatcatc attgacggca cgctatgggg attgatcatc
841 tgtcatcatt acgagccgcg tgccgtgccg atggcgcagc gcgtcgcggc cgaaatgttc
901 gccgacttct tatcgctgca cttcaccgcc gcccaccacc aacgctaagc ggccgcgact
961 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
1021 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
1081 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
1141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
1201 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
1261 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
1321 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
1381 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
1441 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
1501 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
1561 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
1621 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
1681 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
1741 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
1801 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
1861 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
1921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
1981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
2041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
2101 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
2161 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
2221 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
2281 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
2341 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
2401 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
2461 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
2521 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
2581 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
2641 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
2701 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
2761 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
2821 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
2881 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
2941 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
3001 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
3061 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
3121 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
3181 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
3241 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
3301 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
3361 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
3421 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
3481 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
3541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
3601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
3661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
3721 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
3781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
3841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
3901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
3961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
4021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
4081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
4141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
4201 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
4261 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
4321 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
4381 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
4441 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
4501 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
4561 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
4621 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
4681 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
4741 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
4801 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
4861 accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattctgca
4921 gtcgacggta ccgcgggccc gggatcca
//
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