| 质粒名称: | pHAGE-EFS-N22p-3XRFPnls |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 75387 |
| 存储实验室: | Thoru Pederson |
|
载体骨架基本信息 |
|
| 载体名称: | pHAGE-DEST |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6000 bp |
| 完整质粒大小: | 8440 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 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: | N22p |
| 物种来源: | Synthetic |
| 插入大小: | 69 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | EFS |
| 标签/融合蛋白: | 3XmCherry（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Mlu I(未破坏) |
| 3' 克隆位点: | BamH I(未破坏) |
| 5' 测序引物 | cgaaggaatagaagaagaaggtgga |
| 3' 测序引物 | CCAAAGGGAGATCCGACTCG |
|
文献引用方法 |
|
| 材料和方法部分: | pHAGE-EFS-N22p-3XRFPnls was a gift from Thoru Pederson (Addgene plasmid # 75387 ; http://n2t.net/addgene:75387 ; RRID:Addgene_75387) |
| 参考文献部分: | Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T. Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. 10.1038/nbt.3526 PubMed 27088723 |
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 pHAGE-EFS-N22p-3XRFPnls 8440 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION N22p-3XRFPnls.
ACCESSION .
VERSION .
KEYWORDS pHAGE-EFS-N22p-3XRFPnls
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8440)
AUTHORS Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T
TITLE Multiplexed labeling of genomic loci with dCas9 and engineered
sgRNAs using CRISPRainbow.
JOURNAL Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526.
PUBMED 27088723
REFERENCE 2 (bases 1 to 8440)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8440
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2058..2175
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2265..2476
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 2522..2531
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2633..3340
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 3359..4066
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 4085..4792
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 4799..4819
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
misc_feature 4830..5418
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4883..4903)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5301..5312)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5493..5726
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5845..5863)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5931..6035
/gene="bla"
/label=AmpR promoter
CDS 6036..6896
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6254..6273)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7067..7655
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7556..7575
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7809..7826
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 7918..7937
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 8066..8087
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8102..8132
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8140..8156
/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)."
primer_bind 8145..8167
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 8164..8180
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8164..8180
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 8230..8249
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
2221 gctagctgat gcgggccctt cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
2281 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
2341 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
2401 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
2461 tgccgccaga acacaggtgt cgtgacgcgg ttccggttaa ggtgaacgcg tgcaggctgg
2521 cgccaccatg ggtaatgctc ggacccggcg aagagagagg cgggctgaga agcaggcaca
2581 gtggaaggct gcaaacggtg gcggaggtac tagtggctct ggatccggaa gtatggtgag
2641 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
2701 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
2761 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
2821 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
2881 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
2941 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tcactgcagg acggcgagtt
3001 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
3061 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
3121 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
3181 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
3241 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgctga
3301 gggccgccac tccaccggcg gcatggacga gctgtacaag tctggcggcc gctctggcat
3361 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
3421 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
3481 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
3541 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
3601 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
3661 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctcac tgcaggacgg
3721 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
3781 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
3841 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
3901 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
3961 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
4021 cgctgagggc cgccactcca ccggcggcat ggacgagctg tacaagggta gcatcgatgg
4081 tagcatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
4141 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
4201 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
4261 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
4321 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
4381 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctcactgca
4441 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
4501 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
4561 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
4621 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
4681 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
4741 cgaacgcgct gagggccgcc actccaccgg cggcatggac gagctgtaca agctcgagcc
4801 aaagaaaaag cggaaagtgt aatctagata atcaacctct ggattacaaa atttgtgaaa
4861 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
4921 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
4981 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
5041 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
5101 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
5161 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
5221 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
5281 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
5341 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
5401 tttgggccgc ctccccgcct gagatccttt aagaccaatg acttacaagg cagctgtaga
5461 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
5521 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
5581 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
5641 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
5701 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
5761 ataacttgca aagaaatgaa tatcagagag tgagaggccc gggttaatta aggaaagggc
5821 tagatcattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
5881 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
5941 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
6001 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
6061 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
6121 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
6181 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
6241 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
6301 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
6361 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
6421 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
6481 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
6541 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
6601 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
6661 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
6721 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
6781 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
6841 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
6901 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
6961 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
7021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
7081 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
7141 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
7201 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
7261 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
7321 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
7381 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
7441 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
7501 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
7561 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
7621 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
7681 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
7741 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
7801 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
7861 tccccgcgcg ttggccgatt cattaatgca gcaagctcat ggctgactaa ttttttttat
7921 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
7981 ttttggaggc ctaggctttt gcaaaaagct ccccgtggca cgacaggttt cccgactgga
8041 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
8101 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
8161 acacaggaaa cagctatgac atgattacga atttcacaaa taaagcattt ttttcactgc
8221 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
8281 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
8341 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa
8401 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
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