| 质粒名称: | Lentiviral-FOP-dGFP-reporter |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 14885 |
| 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: | 7000 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,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: | mutant LEF-1/TCF-responsive promoter,beta catenin reporter |
| 物种来源: | -- |
| 插入大小: | 100 bp |
| 基因突变: | mutant TCF/LEF binding sites |
| 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-FOP-dGFP-reporter was a gift from Tannishtha Reya (Addgene plasmid # 14885 ; http://n2t.net/addgene:14885 ; RRID:Addgene_14885) |
| 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-FOP-dGFP-reporter 6922 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Lentiviral-FOP-dGFP-reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6922)
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 6922)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6922
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 240..260
/label=T3
/note="T3 promoter, forward primer"
promoter 242..260
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 288..514
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 515..695
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 742..867
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1360..1593
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1778..1822
/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"
regulatory 2156..2165
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 2181..2190
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2187..3032
/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(2232..2253)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2493..2512)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2840..2861
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 3053..3641
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3106..3126)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3524..3535)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3728..3961
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4033..4154
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4070..4089)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4124..4143
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4194..4329
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4256..4275
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4349..4368)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4350..4368)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4378..4395)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4378..4394)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4387..4409)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4536..4991
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4623..4642)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4833..4854
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5017..5121
/gene="bla"
/label=AmpR promoter
CDS 5122..5982
/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(5340..5359)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6153..6741
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6642..6661
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6895..6912
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
1981 aaccccgagg ggacccgaca tagcccgggc tcgagcagct gaagcttgca tgcctgcagg
2041 gccaaagggg gtaaggccaa agggggtaag gccaaaggtc tagagggtat ataatggatc
2101 cggtactcga gatctgcgat ctaagtaagc ttggcattcc ggtactgttg gtaaagccac
2161 catgggatga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
2221 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
2281 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
2341 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
2401 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
2461 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
2521 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
2581 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
2641 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
2701 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
2761 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
2821 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
2881 tcggcatgga cgagctgtac aagaagctta gccatggctt cccgccggag gtggaggagc
2941 aggatgatgg cacgctgccc atgtcttgtg cccaggagag cgggatggac cgtcaccctg
3001 cagcctgtgc ttctgctagg atcaatgtgt aggcggccgc ctgcaggtcg acaatcaacc
3061 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
3121 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
3181 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
3241 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
3301 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
3361 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
3421 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
3481 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
3541 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
3601 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta
3661 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
3721 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
3781 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
3841 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
3901 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
3961 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
4021 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
4081 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
4141 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
4201 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
4261 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
4321 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
4381 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
4441 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
4501 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
4561 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
4621 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
4681 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
4741 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
4801 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
4861 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
4921 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
4981 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
5041 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
5101 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
5161 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
5221 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
5281 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
5341 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
5401 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
5461 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
5521 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
5581 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
5641 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
5701 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
5761 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
5821 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
5881 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
5941 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
6001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
6061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
6121 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
6181 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
6241 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
6301 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
6361 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
6421 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
6481 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
6541 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
6601 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
6661 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
6721 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
6781 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
6841 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
6901 tcagtgagcg aggaagcgga ag
//
