| 质粒名称: | pGreen_dualluc_3'UTR_sensor |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 55206 |
| 存储实验室: | Michael Axtell |
|
载体骨架基本信息 |
|
| 载体名称: | pGreen II |
| 载体出品公司: | pGreen II |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 2500 bp |
| 完整质粒大小: | 7070 bp |
| 载体修饰: | -- |
| 载体类型: | 荧光素酶；植物瞬时表达 |
| 筛选标记: | -- |
| 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: | Renilla luciferase,R-Luc |
| 物种来源: | Renilla reniformis |
| 插入大小: | 936 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pGreen_dualluc_3'UTR_sensor was a gift from Michael Axtell (Addgene plasmid # 55206 ; http://n2t.net/addgene:55206 ; RRID:Addgene_55206) |
| 参考文献部分: | Analysis of complementarity requirements for plant MicroRNA targeting using a Nicotiana benthamiana quantitative transient assay. Liu Q, Wang F, Axtell MJ. Plant Cell. 2014 Feb;26(2):741-53. doi: 10.1105/tpc.113.120972. Epub 2014 Feb 7. 10.1105/tpc.113.120972 PubMed 24510721 |
买家导航
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 pGreen_dualluc_3'UTR_sensor 7069 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Dual-luciferase reporter for microRNA targeting in plants.
ACCESSION .
VERSION .
KEYWORDS pGreen_dualluc_3'UTR_sensor
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7069)
AUTHORS Liu Q, Wang F, Axtell MJ
TITLE Analysis of complementarity requirements for plant MicroRNA
targeting using a Nicotiana benthamiana quantitative transient
assay.
JOURNAL Plant Cell. 2014 Feb;26(2):741-53. doi: 10.1105/tpc.113.120972. Epub
2014 Feb 7.
PUBMED 24510721
REFERENCE 2 (bases 1 to 7069)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7069
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 542..564
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA
(truncated)"
terminator 587..839
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(854..1789)
/codon_start=1
/product="luciferase from the anthozoan coelenterate
Renilla reniformis (sea pansy)"
/label=Rluc
/translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
promoter complement(1823..2002)
/label=NOS promoter
/note="nopaline synthase promoter"
primer_bind 2234..2256
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2248..2265
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2249..2265
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2275..2294
/label=T7
/note="T7 promoter, forward primer"
promoter 2275..2293
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2368..2711
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 2656..2676
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 2718..4370
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(2784..2802)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
promoter complement(4668..4686)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4707..4723)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4707..4723)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4720..4742)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4731..4747
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4755..4786)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4801..4822
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4939..4956)
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 5061..5085
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
rep_origin complement(5176..5764)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5256..5275)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5935..6750)
/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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 6658..6677
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 6742..6761
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 7041..407
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"
ORIGIN
1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgccag cgcctttgcc
421 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
481 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagagatc
541 ttggcaggat atattgtggt gtaacgtatc agcttgcatg ccggtcgatc tagtaacata
601 gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg
661 tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg
721 cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatc
781 gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct
841 gcttgactct agattattgt tcatttttga gaactcgctc aacgaacgat ttgatatatt
901 ttcccatttc atcaggtgca tcttcttgcg aaaaatgaag accttttact ttgacaaatt
961 cagtattagg aaacttcttg gcaccttcaa caatagcatt ggaaaagaat cctgggtccg
1021 attcaataaa catttttggt aaatcatcac ttgcacgtag ataagcatta taattcctaa
1081 caatttgtac aacgtcaggt ttaccacctt ttactaacgg gatttcacga ggccatgata
1141 atgttggacg acgaacttca cctttctctt tgaatggttc aagatatgct gcaaattctt
1201 ctggttctaa ctttctcatg atttttgatg gcaacatggt ttccacgaag aagttattct
1261 ccaaaaccat tttttctcct tcttcagatt tgatcaacgc aatatcttct tcaatatcag
1321 gccattcatc ccatgattca atcacatcta ctacactttc agcgtgaact attgctttga
1381 tcttatcttg atgctcatag ctataatgaa atgccaaaca agcaccccaa tcatggccga
1441 caaaaatgat cttctttggt aaattaagaa gttcaaacca tgcagtaaga tatttgtaat
1501 gatcaagtaa cctataagaa ccattaccag atttgcctga tttgcccata ccaataaggt
1561 ctggtataat acaccgcgct actggctcaa tatgtggcac aacatgtcgc cataaataag
1621 aagaggccgc gttaccatgt aaaaaaataa cagcattttc tgcatgtttt tctgaatcat
1681 aataattaat aaatgaatca agaacattca tttgtttaca tctggcccac cactgcggac
1741 cagttatcat ccgtttcctt tgttctggat cataaacttt cgaagtcatg tcgacctcga
1801 gttgagagtg aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg
1861 agcatttttg acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg
1921 caataatggt ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt
1981 ggctccttca acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat
2041 cggcgggggt cataacgtga ctcccttaat tctccgctca tgatcgataa cattaacgct
2101 tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
2161 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
2221 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
2281 gactcactat agggcgaatt gggtaccggg cccacccccc tactccaaaa atgtcaaaga
2341 tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa tttcgggaaa
2401 cctcctcgga ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga
2461 aggtggctcc tacaaatgcc atcattgcga taaaggaaag gctatcattc aagatgcctc
2521 tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga
2581 cgttccaacc acgtcttcaa agcaagtgga ttgatgtgac atctccactg acgtaaggga
2641 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gtcatttcat
2701 ttggagagga cctcgagatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct
2761 atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc
2821 tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg
2881 agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa
2941 atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg
3001 cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc
3061 tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa
3121 aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta
3181 aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg
3241 gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga
3301 tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg
3361 cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata
3421 ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat
3481 atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga
3541 ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct
3601 tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg
3661 gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag
3721 gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg
3781 gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg
3841 atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc
3901 ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg
3961 atggatggct acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg
4021 ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg
4081 aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg
4141 atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg
4201 aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag
4261 gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa
4321 tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa gaattcctgc
4381 agtcgccatg cggtcattac actatcccta ggatcggtat ccagctagta caaaccggtt
4441 ttactatctt ttactcttga actagttaat gtgtgagtag tttcccgata agggaaatta
4501 gggttcttat agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg
4561 tatttgtaaa atacttctat caataaaatt tctaattcct aaaaccaaaa tccagtacta
4621 aaatccagat cgcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg
4681 ttaattccga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
4741 ctcacaattc cacacaacat acgagccgga aghcataaag tgtaaagcct ggggtgccta
4801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
4861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
5041 aggaaagaac atgaaggcct tgacaggata tattggcggg taaactaagt cgctgtatgt
5101 gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
5341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
5581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
5641 gctgaagcca gttaccttcg gaagaagagt tggtagctct tgatccggca aacaaaccac
5701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
5761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
5821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
5881 aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa cattggtcta gtgattagaa
5941 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
6001 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
6061 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
6121 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
6181 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
6241 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
6301 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
6361 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
6421 taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg
6481 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
6541 gaccatctca tctgtaacaa cattggcaac gctacctttg ccatgtttca gaaacaactc
6601 tggcgcatcg ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc
6661 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga
6721 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
6781 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
6841 ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat
6901 cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa
6961 ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga
7021 tggggcgatt caggcgatcc ccatccaaca gcccgccgtc gagcgggct
//
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