| 质粒名称: | ISceI-GR-RFP<质粒图谱.png/td> |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 17654 |
| 存储实验室: | Tom Misteli |
|
载体骨架基本信息 |
|
| 载体名称: | pDsRed-Monomer-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 6276 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin |
| 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: | I-SceI-GRLBD / SCEI/ GR ligand binding domain / glucocorticoid receptor |
| 物种来源: | R. norvegicus (rat), S. cerevisiae (budding yeast) |
| 基因突变: | C655G mutation in the glucocorticoid receptor (see comments) |
| GenBank编号 | JN247409 |
| 启动子: | CMV |
| 标签/融合蛋白: | mRFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SacI (not destroyed) |
| 3’克隆位点: | BamHI (not destroyed) |
| 5’测序引物 | DsRed1-C |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: |
ISceI-GR-RFP was a gift from Tom Misteli (Addgene plasmid # 17654 ; http://n2t.net/addgene:17654 ; RRID:Addgene_17654)
|
| 参考文献部分: | Positional stability of single double-strand breaks in mammalian cells. Soutoglou E, Dorn JF, Sengupta K, Jasin M, Nussenzweig A, Ried T, Danuser G, Misteli T. Nat Cell Biol. 2007 Jun . 9(6):675-82. 10.1038/ncb1591 PubMed 17486118 |
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 ISceI-GR-RFP 6276 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS ISceI-GR-RFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6276)
AUTHORS Soutoglou E, Dorn JF, Sengupta K, Jasin M, Nussenzweig A, Ried T,
Danuser G, Misteli T
TITLE Positional stability of single double-strand breaks in mammalian
cells.
JOURNAL Nat Cell Biol. 2007 Jun . 9(6):675-82.
PUBMED 17486118
REFERENCE 2 (bases 1 to 6276)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6276
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 614..623
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 620..1294
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Strongin et al., 2007)"
/label=DsRed-Monomer
/note="mammalian codon-optimized"
/translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
GSQ"
primer_bind complement(800..820)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 1018..1037
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 3071..3192
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3108..3127)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3162..3181
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3199..3654)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3336..3357)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3548..3567
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3681..3785
/gene="bla"
/label=AmpR promoter
promoter 3787..4144
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3810..3830)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3995..4130
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4057..4076
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4179..4973
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4233..4252)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4843..4862
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5161..5180)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5205..5252
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5581..6169
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6070..6089
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gtcgccacca tggacaacac cgaggacgtc atcaaggagt tcatgcagtt
661 caaggtgcgc atggagggct ccgtgaacgg ccactacttc gagatcgagg gcgagggcga
721 gggcaagccc tacgagggca cccagaccgc caagctgcag gtgaccaagg gcggccccct
781 gcccttcgcc tgggacatcc tgtcccccca gttccagtac ggctccaagg cctacgtgaa
841 gcaccccgcc gacatccccg actacatgaa gctgtccttc cccgagggct tcacctggga
901 gcgctccatg aacttcgagg acggcggcgt ggtggaggtg cagcaggact cctccctgca
961 ggacggcacc ttcatctaca aggtgaagtt caagggcgtg aacttccccg ccgacggccc
1021 cgtaatgcag aagaagactg ccggctggga gccctccacc gagaagctgt acccccagga
1081 cggcgtgctg aagggcgaga tctcccacgc cctgaagctg aaggacggcg gccactacac
1141 ctgcgacttc aagaccgtgt acaaggccaa gaagcccgtg cagctgcccg gcaaccacta
1201 cgtggactcc aagctggaca tcaccaacca caacgaggac tacaccgtgg tggagcagta
1261 cgagcacgcc gaggcccgcc actccggctc ccagtccgga ctcagatctc gagctcaaat
1321 gaaaaacatc aaaaaaaacc aggtaatgaa cctgggtccg aactctaaac tgctgaaaga
1381 atacaaatcc cagctgatcg aactgaacat cgaacagttc gaagcaggta tcggtctgat
1441 cctgggtgat gcttacatcc gttctcgtga tgaaggtaaa acctactgta tgcagttcga
1501 gtggaaaaac aaagcataca tggaccacgt atgtctgctg tacgatcagt gggtactgtc
1561 cccgccgcac aaaaaagaac gtgttaacca cctgggtaac ctggtaatca cctggggcgc
1621 ccagactttc aaacaccaag ctttcaacaa actggctaac ctgttcatcg ttaacaacaa
1681 aaaaaccatc ccgaacaacc tggttgaaaa ctacctgacc ccgatgtctc tggcatactg
1741 gttcatggat gatggtggta aatgggatta caacaaaaac tctaccaaca aatcgatcgt
1801 actgaacacc cagtctttca ctttcgaaga agtagaatac ctggttaagg gtctgcgtaa
1861 caaattccaa ctgaactgtt acgtaaaaat caacaaaaac aaaccgatca tctacatcga
1921 ttctatgtct tacctgatct tctacaacct gatcaaaccg tacctgatcc cgcagatgat
1981 gtacaaactg ccgaacacta tctcctccga aactttcctg aaatgtcgac ggtaccgcgg
2041 gtatcggaaa tgtcttcagg ctggaatgaa ccttgaagct cgaaaaacaa agaaaaaaat
2101 caaagggatt cagcaagcca ctgcaggagt ctcacaagac acttcggaaa atcctaacaa
2161 aacaatagtt cctgcagcat taccacagct cacccctacc ttggtgtcac tgctggaggt
2221 gattgaaccc gaggtgttgt atgcaggata tgatagctct gttccagatt cagcatggag
2281 aattatgacc acactcaaca tgttaggtgg gcgtcaagtg attgcagcag tgaaatgggc
2341 aaaggcgata ccaggcttca gaaacttaca cctggatgac caaatgaccc tgctacagta
2401 ctcatggatg tttctcatgg catttgccct gggttggaga tcatacagac aatcaagtgg
2461 aaacctgctc tgctttgctc ctgatctgat tattaatgag cagagaatgt ctctacccgg
2521 catgtatgac caatgtaaac acatgctgtt tgtctcctct gaattacaaa gattgcaggt
2581 atcctatgaa gagtatctct gtatgaaaac cttactgctt ctctcctcag ttcctaagga
2641 aggtctgaag agccaagagt tatttgatga gattcgaatg acttatatca aagagctagg
2701 aaaagccatc gtcaaaaggg aagggaactc cagtcagaac tggcaacggt tttaccaact
2761 gacaaagctt ctggactcca tgcatgaggt ggttgagaat ctccttacct actgcttcca
2821 gacatttttg gataagacca tgagtattga attcccagag atgttagctg aaatcatcac
2881 taatcagata ccaaaatatt caaatggaaa tatcaaaaag cttctgtttc atcaaaaatt
2941 tggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
3001 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
3061 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
3121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
3181 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
3241 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
3301 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
3361 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
3421 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
3481 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
3541 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
3601 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
3661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
3721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
3781 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
3841 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
3901 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
3961 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
4021 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
4081 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
4141 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
4201 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
4261 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
4321 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
4381 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
4441 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
4501 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
4561 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
4621 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
4681 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
4741 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
4801 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
4861 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
4921 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
4981 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
5041 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
5101 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
5161 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
5221 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
5281 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
5341 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
5401 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
5461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
5521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
5581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
5641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
5701 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
5761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
5821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
5881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
5941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
6001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
6061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
6121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
6181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
6241 gttatcccct gattctgtgg ataaccgtat taccgc
//
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