| 质粒名称: | Lentiviral-TOP-dGFP-reporter |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 14715 |
| 存储实验室: | Tannishtha Reya |
|
载体骨架基本信息 |
|
| 载体名称: | pRRL.sin-18.ppt |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 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: | LEF-1/TCF-responsive promoter,beta catenin reporter |
| 物种来源: | -- |
| 插入大小: | 100 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | d2 eGFP（主干上的C端） |
| 克隆方法: | 限制性内切酶 |
| 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: | Lentiviral-TOP-dGFP-reporter was a gift from Tannishtha Reya (Addgene plasmid # 14715 ; http://n2t.net/addgene:14715 ; RRID:Addgene_14715) |
| 参考文献部分: | A role for Wnt signalling in self-renewal of haematopoietic stem cells. Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, Hintz L, Nusse R, Weissman IL. Nature. 2003 May 22. 423(6938):409-14. 10.1038/nature01593 PubMed 12717450 |
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 Lentiviral-TOP-dGFP-reporter 7151 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Beta-catenin reporter - 3rd generation self-inactivating lentiviral
plasmid LEF-1/TCF-responsive promoter driven d2-eGFP (destabilized,
half-life of 2h)..
ACCESSION .
VERSION .
KEYWORDS Lentiviral-TOP-dGFP-reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7151)
AUTHORS Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, Hintz
L, Nusse R, Weissman IL
TITLE A role for Wnt signalling in self-renewal of haematopoietic stem
cells.
JOURNAL Nature. 2003 May 22. 423(6938):409-14.
PUBMED 12717450
REFERENCE 2 (bases 1 to 7151)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7151
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 15..603
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 504..523
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 757..774
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 891..912
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 927..957
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 965..981
/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 970..992
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 989..1005
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 989..1005
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 1024..1044
/label=T3
/note="T3 promoter, forward primer"
promoter 1026..1044
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1072..1298
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 1299..1479
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1526..1651
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2144..2377
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2562..2606
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2873..2990
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
regulatory 3169..3178
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 3194..3203
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3200..4045
/codon_start=1
/product="EGFP destabilized by fusion to residues 422-461
of mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours"
/label=d2EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
"
primer_bind complement(3245..3266)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3506..3525)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3853..3874
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 4066..4654
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4119..4139)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4537..4548)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4741..4974
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5046..5167
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5083..5102)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5137..5156
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5207..5342
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5269..5288
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(5362..5381)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5363..5381)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5391..5408)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5391..5407)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5400..5422)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5549..6004
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5636..5655)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5846..5867
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 6030..6134
/gene="bla"
/label=AmpR promoter
CDS 6135..6995
/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(6353..6372)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
721 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
781 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
841 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
901 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
961 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
1021 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
1081 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
1141 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
1201 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
1261 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
1321 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
1381 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
1441 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
1501 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
1561 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
1621 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
1681 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
1741 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
1801 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
1861 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
1921 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
1981 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
2041 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
2101 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
2161 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
2221 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
2281 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
2341 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
2401 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
2461 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
2521 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
2581 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
2641 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
2701 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
2761 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
2821 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa
2881 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
2941 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt atcgatcacg
3001 agactagcct cgatagcccg ggctcgagca gctgaagctt gcatgcctgc aggatcaaag
3061 ggggtaagat caaagggggt aagatcaaag gtctagaggg tatataatgg atccggtact
3121 cgagatctgc gatctaagta agcttggcat tccggtactg ttggtaaagc caccatggga
3181 tgatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
3241 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
3301 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
3361 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
3421 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
3481 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
3541 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
3601 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
3661 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
3721 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
3781 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
3841 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
3901 ggacgagctg tacaagaagc ttagccatgg cttcccgccg gaggtggagg agcaggatga
3961 tggcacgctg cccatgtctt gtgcccagga gagcgggatg gaccgtcacc ctgcagcctg
4021 tgcttctgct aggatcaatg tgtaggcggc cgcctgcagg tcgacaatca acctctggat
4081 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
4141 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
4201 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
4261 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
4321 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
4381 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
4441 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
4501 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
4561 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
4621 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacctttaa
4681 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
4741 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
4801 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
4861 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
4921 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
4981 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
5041 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
5101 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
5161 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
5221 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
5281 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
5341 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
5401 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
5461 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
5521 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
5581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
5641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
5701 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
5761 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
5821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
5881 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
5941 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
6001 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
6061 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
6121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
6181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
6241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
6301 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
6361 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
6421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
6481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
6541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
6601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
6661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
6721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
6781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
6841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
6901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
6961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
7021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
7081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
7141 ccgtagaaaa g
//
