| 质粒名称: | pNLS-iRFP713 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 45468 |
| 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 |
| 空载体大小: | 4055bp |
| 完整质粒大小: | 5006 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 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: | iRFP713 |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 951 bp |
| 基因突变: | T2A, S13L, A92T, V104T, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T, N290Y relative to RpBphP2 |
| GenBank编号 | JN247409 |
| 启动子: | CMV |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 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: | |
| 参考文献部分: | |
买家导航
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 pNLS-iRFP713 5006 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pNLS-iRFP713
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5006)
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 5006)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5006
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 716..725
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 722..1672
/codon_start=1
/product="phytochrome-based near-infrared fluorescent
protein, originally termed iRFP (Filonov et al., 2011)"
/label=iRFP713
/note="name was changed to iRFP713 (Shcherbakova and
Verkhusha, 2013)"
/translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
polyA_signal 1794..1915
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1831..1850)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1885..1904
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1922..2377)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2059..2080)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2271..2290
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2404..2508
/gene="bla"
/label=AmpR promoter
promoter 2510..2867
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2533..2553)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2718..2853
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2780..2799
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2902..3696
/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(2956..2975)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3566..3585
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3884..3903)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3928..3975
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4304..4892
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4793..4812
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttcga attcgccacc atgtcgcgga ggcggcacag
661 ctacgagaac gatggtggac aacctcacaa aaggaggaag acgtctccac cggtcgccac
721 catggcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg atgagccgat
781 ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg ccgacatgac
841 gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg gcgccctgat
901 cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga cgatcgcctt
961 ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga tgcgaaagga
1021 cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg agctcgagcc
1081 tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca gcgccatccg
1141 ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc aagaggtgcg
1201 gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact tcagcggcga
1261 agtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc actatcctgc
1321 ctcaaccgtg ccggcgcagg cccgtcggct ctataccatc aacccggtac ggatcattcc
1381 cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca ccgggcggcc
1441 gattgatctt agcttcgcca tcctgcgcag cgtctcgccc gtccatctgg agttcatgcg
1501 caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc gactgtgggg
1561 attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc gccaagcctg
1621 cgagctagtc gcccaggttc tggcctggca gatcggcgtg atggaagagt gagcggccgc
1681 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
1741 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
1801 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
1861 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
1921 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
1981 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
2041 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
2101 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
2161 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
2221 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
2281 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
2341 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
2401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
2461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
2521 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
2581 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
2701 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
2761 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
2821 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
2881 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
2941 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
3001 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
3061 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
3121 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
3181 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
3241 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
3301 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
3361 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
3421 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
3481 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
3541 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
3601 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
3661 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
3721 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
3781 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
3841 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
3901 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
3961 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
4021 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
4081 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
4141 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
4201 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
4261 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
4321 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
4381 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
4441 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
4501 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
4561 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
4621 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
4681 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
4741 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
4801 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
4861 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
4921 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
4981 tggataaccg tattaccgcc atgcat
//
