| 质粒名称: | pF141 pAcGFP1 SOD1WT |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 26402 |
| 存储实验室: | Elizabeth Fisher |
|
载体骨架基本信息 |
|
| 载体名称: | pAcGFP1-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 5193 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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 (N 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 |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pF141 pAcGFP1 SOD1WT was a gift from Elizabeth Fisher (Addgene plasmid # 26402 ; http://n2t.net/addgene:26402 ; RRID:Addgene_26402) |
| 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 pF141 pAcGFP1 SOD1WT 5193 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pF141 pAcGFP1 SOD1WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5193)
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 5193)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5193
/organism="synthetic DNA construct"
/mol_type="other DNA"
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"
regulatory 547..556
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 553..1269
/codon_start=1
/product="Aequorea coerulescens GFP"
/label=AcGFP1
/note="mammalian codon-optimized"
/translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
GFVTAAAITHGMDELYK"
polyA_signal 1921..2042
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1958..1977)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2012..2031
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2049..2504)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2186..2207)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2398..2417
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2531..2635
/gene="bla"
/label=AmpR promoter
promoter 2637..2994
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2660..2680)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2845..2980
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2907..2926
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3029..3823
/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(3083..3102)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3693..3712
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4011..4030)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4055..4102
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4431..5019
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4920..4939
/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 ccggtcgcca ccatggtgag caagggcgcc gagctgttca ccggcatcgt gcccatcctg
601 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc
661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg
721 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc
781 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag
841 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag
901 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac
961 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac
1021 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc
1081 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg
1141 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc
1201 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag
1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attccatggc gacgaaggcc
1321 gtgtgcgtgc tgaagggcga cggcccagtg cagggcatca tcaatttcga gcagaaggaa
1381 agtaatggac cagtgaaggt gtggggaagc attaaaggac tgactgaagg cctgcatgga
1441 ttccatgttc atgagtttgg agataataca gcaggctgta ccagtgcagg tcctcacttt
1501 aatcctctat ccagaaaaca cggtgggcca aaggatgaag agaggcatgt tggagacttg
1561 ggcaatgtga ctgctgacaa agatggtgtg gccgatgtgt ctattgaaga ttctgtgatc
1621 tcactctcag gagaccattg catcattggc cgcacactgg tggtccatga aaaagcagat
1681 gacttgggca aaggtggaaa tgaagaaagt acaaagacag gaaacgctgg aagtcgtttg
1741 gcttgtggtg taattgggat cgcccaataa gtcgacggta ccgcgggccc gggatccacc
1801 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
1861 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
1921 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
1981 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
2041 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
2101 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
2161 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
2221 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
2281 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
2341 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
2401 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
2461 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
2521 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
2581 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
2641 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
2701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
2761 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
2821 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
2881 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
2941 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
3001 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
3061 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
3121 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
3181 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
3241 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
3301 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
3361 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
3421 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
3481 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
3541 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
3601 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
3661 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
3721 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
3781 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
3841 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
3901 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
3961 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
4021 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
4081 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
4141 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
4201 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
4261 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
4321 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
4381 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
4441 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
4501 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
4561 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
4621 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
4681 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
4741 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
4801 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
4861 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
4921 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
4981 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
5041 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
5101 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
5161 gtcattagtt catagcccat atatggagtt ccg
//
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