| 质粒名称: | pENTR1A-GFP-N2 (FR1) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 19364 |
| 存储实验室: | Eric Campeau, Paul Kaufman |
|
载体骨架基本信息 |
|
| 载体名称: | pENTR1A |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 2348 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） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Enhanced Green Fluorescent Protein |
| 物种来源: | -- |
| 插入大小: | 719bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 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: | pENTR1A-GFP-N2 (FR1) was a gift from Eric Campeau & Paul Kaufman (Addgene plasmid # 19364 ; http://n2t.net/addgene:19364 ; RRID:Addgene_19364) |
| 参考文献部分: | A versatile viral system for expression and depletion of proteins in mammalian cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394 |
买家导航
pENTR1A-GFP-N2 (FR1载体质粒基本信息
pENTR1A-GFP-N2 (FR1质粒图谱载体图谱和pENTR1A-GFP-N2 (FR1载体序列质粒序列多克隆位点信息
质粒图谱.png)
pENTR1A-GFP-N2 (FR1质粒载体简介
pENTR1A-GFP-N2 (FR1质粒序列载体序列
LOCUS pENTR1A-GFP-N2 (FR1) 3071 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Gateway entry vector for C-term eGFP fusion, frame N2.
ACCESSION .
VERSION .
KEYWORDS pENTR1A-GFP-N2 (FR1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3071)
AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
Campisi J, Yaswen P, Cooper PK, Kaufman PD.
TITLE A versatile viral system for expression and depletion of proteins in
mammalian cells.
JOURNAL PLoS One. 2009 Aug 6;4(8):e6529.
PUBMED 19657394
REFERENCE 2 (bases 1 to 3071)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3071
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 103..189
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 316..339
/label=pENTR-F
/note="pENTR vectors, forward primer"
protein_bind 358..457
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 486..538
/label=MCS
/note="multiple cloning site"
regulatory 550..559
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 556..1275
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(601..622)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(862..881)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1209..1230
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(1300..1399)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1511..1530)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1522..2331
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1589..1608)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2421..3009
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2910..2929
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
481 tggatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
541 cggccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
721 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
1021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
1081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
1141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
1201 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
1261 gagctgtaca agtaaagcgg ccgcactcga gatatctaga cccagctttc ttgtacaaag
1321 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
1381 aaaataaaat cattatttgc catccagctg cagctctggc ccgtgtctca aaatctctga
1441 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
1501 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
1561 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
1621 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
1681 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
1741 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
1801 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
1861 tcaggtgaaa atattgttga tgcgctggca gtgtccctgc gccggttgca ttcgattcct
1921 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
1981 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
2041 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
2101 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
2161 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
2221 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
2281 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
2341 gttaattggt tgtaacatta ttcagattgg gccccgttcc actgagcgtc agacccggta
2401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
2461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
2521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
2581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
2641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
2701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
2761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
2821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
2881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
2941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
3001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
3061 gctcacatgt t
//
