| 质粒名称: | pGreen_GUS_competitor |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 55208 |
| 存储实验室: | Michael Axtell |
|
载体骨架基本信息 |
|
| 载体名称: | pGreen II 0229 |
| 载体出品公司: | pGreen II |
| 空载体大小: | 2500 bp |
| 完整质粒大小: | 6850 bp |
| 载体修饰: | -- |
| 载体类型: | 植物表达 |
| 筛选标记: | Basta |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | beta-glucuronidase,GUS |
| 物种来源: | -- |
| 插入大小: | 1812 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_GUS_competitor was a gift from Michael Axtell (Addgene plasmid # 55208 ; http://n2t.net/addgene:55208 ; RRID:Addgene_55208) |
| 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_GUS_competitor 6849 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION GUS-based microRNA target competitor used for testing function as
target mimicry.
ACCESSION .
VERSION .
KEYWORDS pGreen_GUS_competitor
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6849)
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 6849)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6849
/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 588..840
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(860..1411)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(1453..1632)
/label=NOS promoter
/note="nopaline synthase promoter"
primer_bind 1864..1886
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1878..1895
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1879..1895
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1905..1924
/label=T7
/note="T7 promoter, forward primer"
promoter 1905..1923
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1998..2342
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 2286..2306
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 2349..4160
/codon_start=1
/gene="uidA (or gusA) from E. coli"
/product="beta-glucuronidase"
/label=GUS
/translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
primer_bind complement(4449..4469)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(4449..4467)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4488..4504)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4488..4504)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4501..4523)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4512..4528
/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(4536..4566)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4581..4602
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4719..4736)
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4841..4865
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
rep_origin complement(4956..5544)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5036..5055)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5715..6530)
/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 6438..6457
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 6522..6541
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 6821..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 gtaacgttat cagcttgcat gccggtcgat ctagtaacat
601 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
661 gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat
721 gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat
781 cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc
841 tgcttgactc taggggtcat cagatttcgg tgacgggcag gaccggacgg ggcggcaccg
901 gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg
961 ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt
1021 cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca
1081 gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg gagacgtaca
1141 cggtcgactc ggccgtccag tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcga
1201 tgccggcgac ctcgccgtcc acctcggcga cgagccaggg atagcgctcc cgcagacgga
1261 cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt acggaagttg accgtgcttg
1321 tctcgatgta gtggttgacg atggtgcaga ccgccggcat gtccgcctcg gtggcacggc
1381 ggatgtcggc cgggcgtcgt tctgggctca tggtagatcc cctcgatcga gttgagagtg
1441 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg
1501 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
1561 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca
1621 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
1681 cataacgtga ctcccttaat tctccgctca tgatcgataa cattaacgct tacaatttcc
1741 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
1801 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
1861 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
1921 agggcgaatt gggtaccggg cccacccccc tactccaaaa atgtcaaaga tacagtctca
1981 gaagaccaaa gggctattga gacttttcaa caaagggtaa tttcgggaaa cctcctcgga
2041 ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc
2101 tacaaatgcc atcattgcga taaaggaaag gctatcattc aagatgcctc tgccgacagt
2161 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc
2221 acgtcttcaa agcaagtgga ttgatgtgac atctccactg acgtaaggga tgacgcacaa
2281 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg
2341 acctcgagat ggtacgtcct gtagaaaccc caacccgtga aatcaaaaaa ctcgacggcc
2401 tgtgggcatt cagtctggat cgcgaaaact gtggaattga tcagcgttgg tgggaaagcg
2461 cgttacaaga aagccgggca attgctgtgc caggcagttt taacgatcag ttcgccgatg
2521 cagatattcg taattatgcg ggcaacgtct ggtatcagcg cgaagtcttt ataccgaaag
2581 gttgggcagg ccagcgtatc gtgctgcgtt tcgatgcggt cactcattac ggcaaagtgt
2641 gggtcaataa tcaggaagtg atggagcatc agggcggcta tacgccattt gaagccgatg
2701 tcacgccgta tgttattgcc gggaaaagtg tacgtatcac cgtttgtgtg aacaacgaac
2761 tgaactggca gactatcccg ccgggaatgg tgattaccga cgaaaacggc aagaaaaagc
2821 agtcttactt ccatgatttc tttaactatg ccggaatcca tcgcagcgta atgctctaca
2881 ccacgccgaa cacctgggtg gacgacatca ccgtggtgac gcatgtcgcg caagactgta
2941 accacgcgtc tgttgactgg caggtggtgg ccaatggtga tgtcagcgtt gaactgcgtg
3001 atgcggatca acaggtggtt gcaactggac aaggcactag cgggactttg caagtggtga
3061 atccgcacct ctggcaaccg ggtgaaggtt atctctatga actgtgcgtc acagccaaaa
3121 gccagacaga gtgtgacatc tacccgcttc gcgtcggcat ccggtcagtg gcagtgaagg
3181 gcgaacagtt cctgattaac cacaaaccgt tctactttac tggctttggt cgtcatgaag
3241 atgcggactt gcgtggcaaa ggattcgata acgtgctgat ggtgcacgac cacgcattaa
3301 tggactggat tggggccaac tcctaccgta cctcgcatta cccttacgct gaagagatgc
3361 tcgactgggc agatgaacat ggcatcgtgg tgattgatga aactgctgct gtcggcttta
3421 acctctcttt aggcattggt ttcgaagcgg gcaacaagcc gaaagaactg tacagcgaag
3481 aggcagtcaa cggggaaact cagcaagcgc acttacaggc gattaaagag ctgatagcgc
3541 gtgacaaaaa ccacccaagc gtggtgatgt ggagtattgc caacgaaccg gatacccgtc
3601 cgcaaggtgc acgggaatat ttcgcgccac tggcggaagc aacgcgtaaa ctcgacccga
3661 cgcgtccgat cacctgcgtc aatgtaatgt tctgcgacgc tcacaccgat accatcagcg
3721 atctctttga tgtgctgtgc ctgaaccgtt attacggatg gtatgtccaa agcggcgatt
3781 tggaaacggc agagaaggta ctggaaaaag aacttctggc ctggcaggag aaactgcatc
3841 agccgattat catcaccgaa tacggcgtgg atacgttagc cgggctgcac tcaatgtaca
3901 ccgacatgtg gagtgaagag tatcagtgtg catggctgga tatgtatcac cgcgtctttg
3961 atcgcgtcag cgccgtcgtc ggtgaacagg tatggaattt cgccgatttt gcgacctcgc
4021 aaggcatatt gcgcgttggc ggtaacaaga aagggatctt cactcgcgac cgcaaaccga
4081 agtcggcggc ttttctgctg caaaaacgct ggactggcat gaacttcggt gaaaaaccgc
4141 agcagggagg caaacaatga gaattcctgc agtcgccatg cggtcattac actatcccta
4201 ggatcggtat gcagaccggt tttactatct tttactcttg aactagttaa tgtgtgagta
4261 gtttcccgat aagggaaatt agggttctta tagggtttcg ctcatgtgtt gagcatataa
4321 gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc
4381 taaaaccaaa atccagtact aaaatccaga tcgcggccgc caccgcggtg gagctccagc
4441 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
4501 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
4561 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
4621 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
4681 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
4741 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
4801 acagaatcag gggataacgc aggaaagaac atgaaggcct tgacaggata tattggcggg
4861 taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag
4921 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
4981 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
5041 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
5101 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
5161 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
5221 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
5281 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
5341 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
5401 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct
5461 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
5521 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
5581 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
5641 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa
5701 cattggtcta gtgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
5761 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
5821 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
5881 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
5941 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
6001 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
6061 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
6121 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
6181 cctgaatcag gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg
6241 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
6301 tccgtcagcc agtttagtct gaccatctca tctgtaacaa cattggcaac gctacctttg
6361 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcggta gattgtcgca
6421 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
6481 gaatttaatc gcggccttga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
6541 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
6601 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
6661 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
6721 caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc
6781 ctcactttct ggctggatga tggggcgatt caggcgatcc ccatccaaca gcccgccgtc
6841 gagcgggct
//
