| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pEarleyGate102； pEarleyGate 102 |
| 质粒类型: | Gateway载体；植物载体；农杆菌双元表达载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | Gateway |
| 启动子: | 35S |
| 载体大小: | 12454 bp |
| 5' 测序引物及序列: | 35S promoter：CTATCCTTCGCAAGACCCTTC |
| 3' 测序引物及序列: | -- |
| 载体标签: | 青色荧光蛋白ECFP、HA (C ter) |
| 载体抗性: | 卡那霉素、氯霉素（仅空载体） |
| 筛选标记: | 草丁膦（磷丝菌素）phosphinothricin、 |
| 克隆菌株: | DB3.1菌株 |
| 宿主细胞（系）: | 植物细胞、农杆菌 |
| 备注: | -- |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pEarleyGate102
- 库存：
20
- 供应商：
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS Exported 12454 bp ds-DNA circular SYN 26-JUL-2016
DEFINITION Gateway(R)-compatible plant transformation vector with CFP and HA
C-terminal tags.
ACCESSION .
VERSION .
KEYWORDS pEarleyGate 102
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12454)
AUTHORS Earley KW, Haag JR, Pontes O, Opper K, Juehne T, Song K, Pikaard CS.
TITLE Gateway-compatible vectors for plant functional genomics and
proteomics.
JOURNAL Plant J. 2006;45:616-29.
PUBMED 16441352
REFERENCE 2 (bases 1 to 12454)
AUTHORS Pikaard Lab
TITLE Direct Submission
JOURNAL Exported Tuesday, July 26, 2016 from SnapGene Viewer 3.1.4
http://www.snapgene.com
COMMENT The sequence was corrected by inserting a G at position 8562.
FEATURES Location/Qualifiers
source 1..12454
/organism="synthetic DNA construct"
/lab_host="Plant Cells"
/mol_type="other DNA"
misc_feature 1..25
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
terminator 111..363
/note="MAS terminator"
/note="mannopine synthase terminator"
CDS complement(373..924)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase草丁膦（磷丝菌素）
乙酰转移酶"
/note="BlpR"
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(930..1310)
/note="MAS promoter"
/note="mannopine synthase promoter (Velten et al., 1984)"
promoter 2306..2651
/note="35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
protein_bind 2663..2782
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 2812..2842
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
CDS 2896..3555
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 3897..4202
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(4243..4367)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 4375..5088
/codon_start=1
/product="enhanced CFP"
/note="ECFP"
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS 5167..5193
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
terminator 5225..5932
/note="OCS terminator"
/note="octopine synthase terminator"
misc_feature 6176..6200
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS 7500..8129
/codon_start=1
/product="stability protein from Pseudomonas plasmid pVS1"
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 8558..9631
/codon_start=1
/product="replication protein from Pseudomonas plasmid
pVS1"
/note="pVS1 RepA"
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 9697..9891
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 10235..10375
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(10561..11149)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11236..12030)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
61 gacgttttta atgtactgaa ttaacgccga attaattcga gctcggatct gataatttat
121 ttgaaaattc ataagaaaag caaacgttac atgaattgat gaaacaatac aaagacagat
181 aaagccacgc acatttagga tattggccga gattactgaa tattgagtaa gatcacggaa
241 tttctgacag gagcatgtct tcaattcagc ccaaatggca gttgaaatac tcaaaccgcc
301 ccatatgcag gagcggatca ttcattgttt gtttggttgc ctttgccaac atgggagtcc
361 aagattctgc agtcaaatct cggtgacggg caggaccgga cggggcggta ccggcaggct
421 gaagtccagc tgccagaaac ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg
481 cagcatgccg cggggggcat atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg
541 cagcccgatg acagcgacca cgctcttgaa gccctgtgcc tccagggact tcagcaggtg
601 ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg ggggagacgt acacggtcga
661 ctcggccgtc cagtcgtagg cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc
721 gacctcgccg tccacctcgg cgacgagcca gggatagcgc tcccgcagac ggacgaggtc
781 gtccgtccac tcctgcggtt cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat
841 gtagtggttg acgatggtgc agaccgccgg catgtccgcc tcggtggcac ggcggatgtc
901 ggccgggcgt cgttctgggc tcatcgattc gatttggtgt atcgagattg gttatgaaat
961 tcagatgcta gtgtaatgta ttggtaattt gggaagatat aataggaagc aaggctattt
1021 atccatttct gaaaaggcga aatggcgtca ccgcgagcgt cacgcgcatt ccgttcttgc
1081 tgtaaagcgt tgtttggtac acttttgact agcgaggctt ggcgtgtcag cgtatctatt
1141 caaaagtcgt taatggctgc ggatcaagaa aaagttggaa tagaaacaga atacccgcga
1201 aattcaggcc cggttgccat gtcctacacg ccgaaataaa cgaccaaatt agtagaaaaa
1261 taaaaactga ctcggatact tacgtcacgt cttgcgcact gatttgaaaa atctcagaat
1321 tccaatccca caaaaatctg agcttaacag cacagttgct cctctcagag cagaatcggg
1381 tattcaacac cctcatatca actactacgt tgtgtataac ggtccacatg ccggtatata
1441 cgatgactgg ggttgtacaa aggcggcaac aaacggcgtt cccggagttg cacacaagaa
1501 atttgccact attacagagg caagagcagc agctgacgcg tacacaacaa gtcagcaaac
1561 agacaggttg aacttcatcc ccaaaggaga agctcaactc aagcccaaga gctttgctaa
1621 ggccctaaca agcccaccaa agcaaaaagc ccactggctc acgctaggaa ccaaaaggcc
1681 cagcagtgat ccagccccaa aagagatctc ctttgccccg gagattacaa tggacgattt
1741 cctctatctt tacgatctag gaaggaagtt cgaaggtgaa ggtgacgaca ctatgttcac
1801 cactgataat gagaaggtta gcctcttcaa tttcagaaag aatgctgacc cacagatggt
1861 tagagaggcc tacgcagcag gtctcatcaa gacgatctac ccgagtaaca atctccagga
1921 gatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaattgcat
1981 caagaacaca gagaaagaca tatttctcaa gatcagaagt actattccag tatggacgat
2041 tcaaggcttg cttcataaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt
2101 tcctactgaa tctaaggcca tgcatggagt ctaagattca aatcgaggat ctaacagaac
2161 tcgccgtgaa gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga
2221 aaatcttcgt caacatggtg gagcacgaca ctctggtcta ctccaaaaat gtcaaagata
2281 cagtctcaga agaccaaagg gctattgaga cttttcaaca aaggataatt tcgggaaacc
2341 tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag
2401 gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa gatctctctg
2461 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
2521 ttccaaccac gtcttcaaag caagtggatt gatgtgacat ctccactgac gtaagggatg
2581 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
2641 tggagaggac acgctcgaga tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
2701 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
2761 taaaacacaa catatccagt catattggcg gccgcattag gcaccccagg ctttacactt
2821 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga
2881 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
2941 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
3001 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
3061 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
3121 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
3181 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
3241 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
3301 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
3361 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
3421 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
3481 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
3541 cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat
3601 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
3661 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
3721 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
3781 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
3841 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
3901 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
3961 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
4021 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
4081 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
4141 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
4201 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt
4261 tgtgttttac agcattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
4321 tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgcc tagggtgagc
4381 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
4441 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
4501 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
4561 accctgacct ggggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
4621 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
4681 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
4741 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
4801 tacaactaca tcagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
4861 gccaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
4921 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
4981 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
5041 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga
5101 tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga
5161 tctagatacc catacgatgt tccagattac gcttaattaa gacccgggac tagtccctag
5221 agtcctgctt taatgagata tgcgagacgc ctatgatcgc atgatatttg ctttcaattc
5281 tgttgtgcac gttgtaaaaa acctgagcat gtgtagctca gatccttacc gccggtttcg
5341 gttcattcta atgaatatat cacccgttac tatcgtattt ttatgaataa tattctccgt
5401 tcaatttact gattgtaccc tactacttat atgtacaata ttaaaatgaa aacaatatat
5461 tgtgctgaat aggtttatag cgacatctat gatagagcgc cacaataaca aacaattgcg
5521 ttttattatt acaaatccaa ttttaaaaaa agcggcagaa ccggtcaaac ctaaaagact
5581 gattacataa atcttattca aatttcaaaa gtgccccagg ggctagtatc tacgacacac
5641 cgagcggcga actaataacg ctcactgaag ggaactccgg ttccccgccg gcgcgcatgg
5701 gtgagattcc ttgaagttga gtattggccg tccgctctac cgaaagttac gggcaccatt
5761 caacccggtc cagcacggcg gccgggtaac cgacttgctg ccccgagaat tatgcagcat
5821 ttttttggtg tatgtgggcc ccaaatgaag tgcaggtcaa accttgacag tgacgacaaa
5881 tcgttgggcg ggtccagggc gaattttgcg acaacatgtc gaggctcagc aggacctgca
5941 ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
6001 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
6061 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgctagagca
6121 gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca gtgtttgaca
6181 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataac ggatatttaa
6241 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
6301 ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag tcggcttctg
6361 acgttcagtg cagccgtctt ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca
6421 ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc gcataaagta
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文献和实验相关实验
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不需要切割带来的二次纯化。4，改良版PET-32a：带His标签（N端）+TrxA(硫氧还蛋白)说明：此载体表达的融合蛋白的N端带有His标签，可以通过一步亲和层析得到纯化，TrxA(硫氧还蛋白)与目的蛋白之间含有thrombin(凝血酶位点)和Linker（102bp）序列。5，改良版PET-NusA：带His标签（N端）+NusA(角质蛋白A)说明：此载体表达的融合蛋白的N端带有His标签，可以通过一步亲和层析得到纯化，NusA(角质蛋白)与目的蛋白之间含有thrombin(凝血
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pEarleyGate102载体
¥2000
