| 质粒名称: | pF146 pSOD1WTAcGFP1 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 26407 |
| 存储实验室: | Elizabeth Fisher |
|
载体骨架基本信息 |
|
| 载体名称: | pAcGFP1-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 5187 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: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | SOD1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 470 bp |
| 标签/融合蛋白: | GFP (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (未破坏) |
| 3’克隆位点: | SalI(未破坏) |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: | pF146 pSOD1WTAcGFP1 was a gift from Elizabeth Fisher (Addgene plasmid # 26407 ; http://n2t.net/addgene:26407 ; RRID:Addgene_26407) |
| 参考文献部分: | Modification of superoxide dismutase 1 (SOD1) properties by a GFP tag--implications for research into amyotrophic lateral sclerosis (ALS). Stevens JC, Chia R, Hendriks WT, Bros-Facer V, van Minnen J, Martin JE, Jackson GS, Greensmith L, Schiavo G, Fisher EM. PLoS One. 2010 . 5(3):e9541. 10.1371/journal.pone.0009541 PubMed 20221404 |
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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 pF146 pSOD1WTAcGFP1 5187 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pF146 pSOD1WTAcGFP1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5187)
AUTHORS Stevens JC, Chia R, Hendriks WT, Bros-Facer V, van Minnen J, Martin
JE, Jackson GS, Greensmith L, Schiavo G, Fisher EM
TITLE Modification of superoxide dismutase 1 (SOD1) properties by a GFP
tag--implications for research into amyotrophic lateral sclerosis
(ALS).
JOURNAL PLoS One. 2010 . 5(3):e9541.
PUBMED 20221404
REFERENCE 2 (bases 1 to 5187)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5187
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 11..599
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 500..519
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 774..1077
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1078..1281
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1232..1252
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 1847..2566
/codon_start=1
/product="Aequorea coerulescens GFP"
/label=AcGFP1
/note="mammalian codon-optimized"
/translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
GFVTAAAITHGMDELYK"
polyA_signal 2688..2809
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2725..2744)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2779..2798
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2816..3271)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2953..2974)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3165..3184
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3298..3402
/gene="bla"
/label=AmpR promoter
promoter 3404..3761
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3427..3447)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3612..3747
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3674..3693
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3796..4590
/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(3850..3869)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4460..4479
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4778..4797)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4822..4869
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
721 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
781 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
841 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
901 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
961 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
1021 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
1081 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
1141 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
1201 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
1261 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
1321 cagatctcga gctcaagctt cgaattccat ggcgacgaag gccgtgtgcg tgctgaaggg
1381 cgacggccca gtgcagggca tcatcaattt cgagcagaag gaaagtaatg gaccagtgaa
1441 ggtgtgggga agcattaaag gactgactga aggcctgcat ggattccatg ttcatgagtt
1501 tggagataat acagcaggct gtaccagtgc aggtcctcac tttaatcctc tatccagaaa
1561 acacggtggg ccaaaggatg aagagaggca tgttggagac ttgggcaatg tgactgctga
1621 caaagatggt gtggccgatg tgtctattga agattctgtg atctcactct caggagacca
1681 ttgcatcatt ggccgcacac tggtggtcca tgaaaaagca gatgacttgg gcaaaggtgg
1741 aaatgaagaa agtacaaaga caggaaacgc tggaagtcgt ttggcttgtg gtgtaattgg
1801 gatcgcccaa gggtcgacgg taccgcgggc ccgggatcca ccggtcatgg tgagcaaggg
1861 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
1921 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
1981 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
2041 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
2101 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
2161 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
2221 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
2281 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
2341 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
2401 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
2461 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
2521 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtgagcgg ccgcgactct
2581 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
2641 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
2701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
2761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
2821 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
2881 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
2941 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
3001 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
3061 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
3121 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
3181 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
3241 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
3301 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
3361 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
3421 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
3481 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
3541 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
3601 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
3661 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
3721 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
3781 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
3841 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
3901 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
3961 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
4021 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
4081 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
4141 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
4201 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
4261 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
4321 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
4381 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
4441 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
4501 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
4561 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
4621 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
4681 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
4741 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
4801 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
4861 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
4921 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
4981 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
5041 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
5101 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
5161 gagcgtcaga ccccgtagaa aagatca
//
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