| 质粒名称: | pE6c |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 17466 |
| 存储实验室: | Giovanna Benvenuto |
|
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: | 3023 bp |
| 载体修饰: | A short cloning site region and stop codon replaced ccdB gene in pENTR1A. Inserted EYFP tag |
| 载体类型: | 哺乳动物表达，细菌表达，酵母表达，昆虫表达 |
| 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: | None |
| 标签/融合蛋白: | EYFP (C terminal on backbone) |
| 克隆方法: | Gateway Cloning |
| 5' 测序引物 | pENTR-F (5'-CTACAAACTCTTCCTGTTAGTTAG-3') |
| 3' 测序引物 | pENTR-R (5'-ATGGCTCATAACACCCCTTG-3') |
|
文献引用方法 |
|
| 材料和方法部分: | pE6c was a gift from Giovanna Benvenuto (爱迪基因 plasmid # 17466) |
| 参考文献部分: | A modified Gateway cloning strategy for overexpressing tagged proteins in plants. Dubin MJ, Bowler C, Benvenuto G. Plant Methods. 2008 Jan 22. 4(1):3. 10.1186/1746-4811-4-3 PubMed 18211686 |
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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 pE6c 3023 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION modified pENTR vector for C-terminal EYFP tag fusion with your gene
of interest.
ACCESSION .
VERSION .
KEYWORDS pE6c
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3023)
AUTHORS Dubin MJ, Bowler C, Benvenuto G
TITLE A modified Gateway cloning strategy for overexpressing tagged
proteins in plants.
JOURNAL Plant Methods. 2008 Jan 22. 4(1):3.
PUBMED 18211686
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pE6c|
FEATURES Location/Qualifiers
source 1..3023
/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 518..1237
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(563..584)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(824..843)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1171..1192
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(1252..1351)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1463..1482)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1474..2283
/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(1541..1560)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2373..2961
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2862..2881
/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 tggatccggt acagatttcg gacatgcggc cgcacctatg gtgagcaagg gcgaggagct
541 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
601 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
661 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
721 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
781 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
841 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
901 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
961 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
1021 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
1081 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
1141 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
1201 cgggatcact ctcggcatgg acgagctgta caagtaactc gagatatcta gacccagctt
1261 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
1321 tcactatcag tcaaaataaa atcattattt gccatccagc tgcagctctg gcccgtgtct
1381 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
1441 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
1501 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
1561 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
1621 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
1681 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
1741 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
1801 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgtccct gcgccggttg
1861 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
1921 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
1981 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
2041 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
2101 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
2161 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
2221 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
2281 taatcagaat tggttaattg gttgtaacat tattcagatt gggccccgtt ccactgagcg
2341 tcagacccgg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
2401 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
2461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
2521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
2581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
2641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
2701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
2761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
2821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
2881 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
2941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
3001 tgctggcctt ttgctcacat gtt
//
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