| 质粒名称: | pENTR4-GFP-C1 (w392-1) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 17396 |
| 存储实验室: | Eric Campeau, Paul Kaufman |
|
载体骨架基本信息 |
|
| 载体名称: | pENTR4 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 3028bp |
| 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: | GFP |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 716 bp |
| 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: | pENTR4-GFP-C1 (w392-1) was a gift from Eric Campeau & Paul Kaufman (Addgene plasmid # 17396 ; http://n2t.net/addgene:17396 ; RRID:Addgene_17396) |
| 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 |
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pENTR4-GFP-C1 (w392-1)载体质粒基本信息
pENTR4-GFP-C1 (w392-1)质粒图谱载体图谱和pENTR4-GFP-C1 (w392-1)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pENTR4-GFP-C1 (w392-1)质粒载体简介
pENTR4-GFP-C1 (w392-1)质粒序列载体序列
LOCUS pENTR4-GFP-C1 (w392-1) 3028 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Gateway entry vector for N-term eGFP fusion, frame C1.
ACCESSION .
VERSION .
KEYWORDS pENTR4-GFP-C1 (w392-1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3028)
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 3028)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3028
/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"
CDS 463..1179
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(508..529)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(769..788)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1116..1137
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 1180..1245
/label=MCS
/note="multiple cloning site"
protein_bind complement(1257..1356)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1468..1487)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1479..2288
/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
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1546..1565)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2378..2966
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2867..2886
/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 gcaggctcca ccatggtgag caagggcgag
481 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
541 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
601 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
661 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
721 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
781 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
841 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
901 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
961 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
1021 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
1081 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
1141 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagct
1201 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagaccc
1261 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
1321 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg
1381 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
1441 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
1501 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
1561 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
1621 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
1681 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
1741 ctcctggtga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
1801 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
1861 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
1921 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
1981 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
2041 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
2101 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
2161 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
2221 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
2281 ttttctaatc agaattggtt aattggttgt aacattattc agattgggcc ccgttccact
2341 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
2401 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
2461 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
2521 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
2581 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
2641 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
2701 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
2761 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
2821 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
2881 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
2941 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
3001 ccttttgctg gccttttgct cacatgtt
//
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