| 质粒名称: | pLKO.1.sh.beta-catenin.1248 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 19761 |
| 存储实验室: | William Hahn |
|
载体骨架基本信息 |
|
| 载体名称: | pLKO.1 puro |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7032 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,慢病毒,RNAi |
| 筛选标记: | 嘌呤霉素 |
| 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: | small hairpin RNA against beta-catenin,CTNNB |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI（克隆过程中销毁） |
| 3' 克隆位点: | EcoRI（克隆过程中销毁） |
| 5' 测序引物 | LKO.1 5' |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pLKO.1.sh.beta-catenin.1248 was a gift from William Hahn (Addgene plasmid # 19761 ; http://n2t.net/addgene:19761 ; RRID:Addgene_19761) |
| 参考文献部分: | CDK8 is a colorectal cancer oncogene that regulates beta-catenin activity. Firestein R, Bass AJ, Kim SY, Dunn IF, Silver SJ, Guney I, Freed E, Ligon AH, Vena N, Ogino S, Chheda MG, Tamayo P, Finn S, Shrestha Y, Boehm JS, Jain S, Bojarski E, Mermel C, Barretina J, Chan JA, Baselga J, Tabernero J, Root DE, Fuchs CS, Loda M, Shivdasani RA, Meyerson M, Hahn WC. Nature. 2008 Sep 25. 455(7212):547-51. 10.1038/nature07179 PubMed 18794900 |
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 pLKO.1.sh.beta-catenin.1248 7083 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLKO.1.sh.beta-catenin.1248
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7083)
AUTHORS Firestein R, Bass AJ, Kim SY, Dunn IF, Silver SJ, Guney I, Freed E,
Ligon AH, Vena N, Ogino S, Chheda MG, Tamayo P, Finn S, Shrestha Y,
Boehm JS, Jain S, Bojarski E, Mermel C, Barretina J, Chan JA,
Baselga J, Tabernero J, Root DE, Fuchs CS, Loda M, Shivdasani RA,
Meyerson M, Hahn WC
TITLE CDK8 is a colorectal cancer oncogene that regulates beta-catenin
activity.
JOURNAL Nature. 2008 Sep 25. 455(7212):547-51.
PUBMED 18794900
REFERENCE 2 (bases 1 to 7083)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7083
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 50..67
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 184..205
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 220..250
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 258..274
/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 263..285
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 282..298
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 282..298
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 317..337
/label=T3
/note="T3 promoter, forward primer"
promoter 319..337
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 365..591
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 592..772
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 819..944
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1437..1670
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
promoter 2197..2437
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 2197..2217
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2368..2387
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 2541..2658
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2707..3217
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 3136..3154
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 3239..3838
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3239..3258)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3735..3755
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3966..4199
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4271..4392
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4308..4327)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4362..4381
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4432..4567
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4494..4513
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4587..4606)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4588..4606)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4616..4633)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4616..4632)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4625..4647)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4774..5229
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4861..4880)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5071..5092
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5255..5359
/gene="bla"
/label=AmpR promoter
CDS 5360..6220
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5578..5597)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6391..6979
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6880..6899
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
61 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
121 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
181 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
241 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
301 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
361 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
421 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
481 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
541 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
601 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
661 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
721 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
781 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
841 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
901 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
961 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
1021 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
1081 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
1141 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
1201 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
1261 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
1321 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1381 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1441 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1501 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1561 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1621 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1681 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1741 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1801 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1861 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1921 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1981 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
2041 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
2101 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcga
2161 tcacgagact agcctcgagc ggccgccccc ttcaccgagg gcctatttcc catgattcct
2221 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
2281 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
2341 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
2401 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggagg tgctatctgt
2461 ctgctctact cgagtagagc agacagatag cacctttttt aattctcgac ctcgagacaa
2521 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
2581 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
2641 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact ttggccgcgg
2701 ctcgaggggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
2761 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt
2821 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
2881 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
2941 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
3001 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
3061 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
3121 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
3181 ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat
3241 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
3301 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
3361 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
3421 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
3481 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
3541 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
3601 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
3661 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
3721 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
3781 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
3841 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc
3901 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
3961 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
4021 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
4081 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
4141 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
4201 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
4261 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
4321 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
4381 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
4441 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
4501 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
4561 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
4621 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
4681 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
4741 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
4801 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
4861 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
4921 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
4981 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
5041 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
5101 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
5161 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
5221 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
5281 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
5341 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
5401 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
5461 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
5521 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
5581 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
5641 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
5701 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
5761 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
5821 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
5881 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
5941 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
6001 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
6061 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
6121 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
6181 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
6241 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
6301 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
6361 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
6421 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
6481 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
6541 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
6601 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
6661 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
6721 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
6781 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
6841 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
6901 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
6961 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
7021 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
7081 tac
//
