| 质粒名称: | pCytochrome C-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 41182 |
| 存储实验室: | Douglas Green |
|
载体骨架基本信息 |
|
| 载体名称: | pGFP-N1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 5015 bp |
| 载体类型: | 哺乳细胞表达 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cytochrome C,cytog |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 330 bp |
| 启动子 | CMV |
| 标签/融合蛋白: | GFP（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SalI (克隆过程中被销毁) |
| 3' 克隆位点: | BglII (克隆过程中被销毁) |
| 5' 测序引物 | acgtgtcgacctaatatgggtgatgttgaaaaagg |
| 3' 测序引物 | acagatctttctcattagtagcctttttaag |
|
文献引用方法 |
|
| 材料和方法部分: | pCytochrome C-GFP was a gift from Douglas Green (Addgene plasmid # 41182 ; http://n2t.net/addgene:41182 ; RRID:Addgene_41182) |
| 参考文献部分: | The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant. Goldstein JC, Waterhouse NJ, Juin P, Evan GI, Green DR. Nat Cell Biol. 2000 Mar;2(3):156-62. 10.1038/35004029 PubMed 10707086 |
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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 pCytochrome C-GFP 5014 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCytochrome C-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5014)
AUTHORS Goldstein JC, Waterhouse NJ, Juin P, Evan GI, Green DR
TITLE The coordinate release of cytochrome c during apoptosis is rapid,
complete and kinetically invariant.
JOURNAL Nat Cell Biol. 2000 Mar;2(3):156-62.
PUBMED 10707086
REFERENCE 2 (bases 1 to 5014)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5014
/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 954..963
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 960..1679
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1005..1026)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1266..1285)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1613..1634
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1802..1923
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1839..1858)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1893..1912
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1930..2385)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2067..2088)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2279..2298
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2412..2516
/gene="bla"
/label=AmpR promoter
promoter 2518..2875
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2541..2561)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2726..2861
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2788..2807
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2910..3704
/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(2964..2983)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3574..3593
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3892..3911)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3936..3983
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4312..4900
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4801..4820
/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 atctcgacct aatatgggtg atgttgaaaa aggcaagaag atttttgttc
661 agaagtgtgc ccagtgccac actgtggaaa agggaggcaa gcataagact ggaccaaatc
721 tccacggtct gttcgggcgg aagacaggcc aggctgctgg attctcttac acagatgcca
781 acaagaacaa aggcatcacc tggggagagg ataccctgat ggagtatttg gagaatccca
841 aaaagtacat ccctggaaca aaaatgatct tcgctggaat taagaagaag ggagaaaggg
901 cagacctaat agcttatctt aaaaaggcta ctaatgagaa agatccaccg gtcgccacca
961 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
1021 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
1081 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
1141 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
1201 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
1261 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
1321 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
1381 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
1441 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
1501 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
1561 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
1621 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
1681 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
1741 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
1801 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
1861 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
1921 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
1981 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
2041 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
2101 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
2161 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
2221 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
2281 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
2341 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
2401 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
2461 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
2521 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
2581 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
2641 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
2701 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
2761 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
2821 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
2881 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
2941 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
3001 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
3061 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
3121 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
3181 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
3241 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
3301 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
3361 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
3421 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
3481 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
3541 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
3601 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
3661 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
3721 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
3781 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
3841 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
3901 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
3961 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
4021 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
4081 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
4141 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
4201 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
4261 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
4321 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
4381 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
4441 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
4501 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
4561 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
4621 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
4681 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
4741 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
4801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
4861 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
4921 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
4981 tgattctgtg gataaccgta ttaccgccat gcat
//
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