| 质粒名称: | pEB1-sfGFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 103983 |
| 存储实验室: | Philippe Cluzel |
|
载体骨架基本信息 |
|
| 载体名称: | pEB1 |
| 空载体大小: | 3674 bp |
| 完整质粒大小: | 4391bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | MG1655 |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sfGFP |
| 物种来源: | Synthetic |
| 插入大小: | 717 bp |
| 基因突变: | Mutations relative to wild-type GFP (S30R, Y39N, F64L, S65T, F99S, N105T, Y145F, M153T, V163A, I171V, A206V) |
| 启动子: | proC |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | AgeI(未破坏) |
| 5’测序引物 | GCGTATCACGAGGCCCTTTC |
| 3’测序引物 | AAAGGGAAAACTGTCCATATGCAC |
|
文献引用方法 |
|
| 材料和方法部分: |
pEB1-sfGFP was a gift from Philippe Cluzel (Addgene plasmid # 103983 ; http://n2t.net/addgene:103983 ; RRID:Addgene_103983)
|
| 参考文献部分: | Systematic characterization of maturation time of fluorescent proteins in living cells. Balleza E, Kim JM, Cluzel P. Nat Methods. 2018 Jan;15(1):47-51. doi: 10.1038/nmeth.4509. Epub 2017 Nov 20. 10.1038/nmeth.4509 PubMed 29320486 |
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 pEB1-sfGFP 4391 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Plasmid encoding sfGFP.
ACCESSION .
VERSION .
KEYWORDS pEB1-sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4391)
AUTHORS Balleza E, Kim JM, Cluzel P
TITLE Systematic characterization of maturation time of fluorescent
proteins in living cells.
JOURNAL Nat Methods. 2018 Jan;15(1):47-51. doi: 10.1038/nmeth.4509. Epub
2017 Nov 20.
PUBMED 29320486
REFERENCE 2 (bases 1 to 4391)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4391
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 182..895
/codon_start=1
/product="green fluorescent protein"
/label=GFP
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(212..240)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 835..855
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
terminator 936..1022
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1560..2510)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 2558..2780
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
terminator 3272..3366
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3397..4191)
/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(3508..3527)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4118..4137
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4370..4388
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ttcgtcttca ccacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg
61 gttgctggat aactttacgg gcatgcataa ggctcgtatg atatattcag ggagaccaca
121 acggtttccc tctacaaata attttgttta acttttctag atttaagaag gagatataca
181 tatgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
241 tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg caacaaacgg
301 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
361 tgtcactact ttaacttatg gtgttcaatg cttttcaaga tacccagatc atatgaaaca
421 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatatcttt
481 caaagatgac gggacctaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
541 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
601 attggaatac aactttaact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
661 aatcaaagct aacttcaaaa ttagacacaa cgttgaagat ggaagcgttc aactagcaga
721 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
781 cctgtccaca caatctgtcc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
841 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaataaat
901 gtccagacct gcaggcatgc aagctctaga ggcatcaaat aaaacgaaag gctcagtcga
961 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
1021 atccgccgcc ctagacctag ggtacgggtt ttgctgcccg caaacgggct gttctggtgt
1081 tgctagtttg ttatcagaat cgcagatccg gcttcagccg gtttgccggc tgaaagcgct
1141 atttcttcca gaattgccat gattttttcc ccacgggagg cgtcactggc tcccgtgttg
1201 tcggcagctt tgattcgata agcagcatcg cctgtttcag gctgtctatg tgtgactgtt
1261 gagctgtaac aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg
1321 tttcacctgt tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat
1381 ggtgaacagc tttgaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta
1441 caccgttttc atctgtgcat atggacagtt ttccctttga tatgtaacgg tgaacagttg
1501 ttctactttt gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct
1561 cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc
1621 catgagaacg aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca
1681 aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta
1741 tgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg
1801 ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac
1861 gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa
1921 tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt
1981 tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt
2041 ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca
2101 aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa aagataaggc
2161 aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc
2221 cactggaaaa tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc
2281 agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc
2341 tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc
2401 gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca
2461 tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat
2521 tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
2581 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
2641 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
2701 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
2761 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
2821 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
2881 ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag
2941 tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg
3001 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
3061 aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg
3121 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
3181 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
3241 gcttacccgt cttactgtcc ctagtgcttg gattctcacc aataaaaaac gcccggcggc
3301 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
3361 ggagtccaag cgagctctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
3421 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
3481 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
3541 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
3601 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
3661 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
3721 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
3781 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
3841 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
3901 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
3961 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
4021 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
4081 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
4141 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
4201 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
4261 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
4321 caattccgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
4381 cacgaggccc t
//
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