| 质粒名称: | Tcf/Lef:H2B-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 32610 |
| 存储实验室: | Anna-Katerina Hadjantonakis |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | -- |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 5677 bp |
| 载体修饰: | -- |
| 载体类型: | -- |
| 筛选标记: | 新霉素（G418筛选） |
| 载体抗性: | 卡那霉素，50μg/mL |
| 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: | Tcf/Lef:H2B-GFP |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | hsp68 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AseI(未破坏) |
| 3’克隆位点: | SfiI (未破坏) |
| 5’测序引物 | GF PGenotFW: A CAACAAGCGCTCGACCATCAC; |
| 3’测序引物 | GFPGenotRW: AGTCGATGCCCTTCAGCTCGAT |
|
文献引用方法 |
|
| 材料和方法部分: | Tcf/Lef:H2B-GFP was a gift from Anna-Katerina Hadjantonakis (Addgene plasmid # 32610 ; http://n2t.net/addgene:32610 ; RRID:Addgene_32610) |
| 参考文献部分: | A sensitive and bright single-cell resolution live imaging reporter of Wnt/ss-catenin signaling in the mouse. Ferrer-Vaquer A, Piliszek A, Tian G, Aho RJ, Dufort D, Hadjantonakis AK. BMC Dev Biol. 2010 Dec 21;10:121. 10.1186/1471-213X-10-121 PubMed 21176145 |
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 Tcf/Lef:H2B-GFP 5677 bp ds-DNA circular SYN 11-MAY-2023
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Tcf/Lef:H2B-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5677)
AUTHORS Ferrer-Vaquer A, Piliszek A, Tian G, Aho RJ, Dufort D, Hadjantonakis
AK
TITLE A sensitive and bright single-cell resolution live imaging reporter
of Wnt/ss-catenin signaling in the mouse.
JOURNAL BMC Dev Biol. 2010 Dec 21;10:121.
PUBMED 21176145
REFERENCE 2 (bases 1 to 5677)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 11, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5677
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 81..438
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(104..124)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 289..424
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 351..370
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 473..1267
/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(527..546)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1137..1156
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(1455..1474)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 1499..1546
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 1875..2463
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2364..2383
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2589..2606
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 2826..2842
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 2827..2843
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 3804..4181
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAVTKYTSAK"
regulatory 4194..4203
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4200..4919
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 4200..4916
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4245..4266)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4506..4525)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4853..4874
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 5042..5163
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5079..5098)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5133..5152
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(5170..5625)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5307..5328)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5519..5538
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5652..79
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
61 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
121 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
181 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
241 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
301 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
361 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
421 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
481 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
541 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
601 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
661 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
721 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
781 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
841 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
901 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
961 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
1021 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
1081 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
1141 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
1201 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
1261 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
1321 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
1381 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
1441 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
1501 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
1561 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
1621 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
1681 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
1741 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
1801 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
1861 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
1921 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
1981 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
2041 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
2101 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
2161 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
2221 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
2281 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
2341 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
2401 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
2461 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
2521 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
2581 ttattaatat taaccctcac taaagggaac aaaagctggt accgggcccc ccctcgagta
2641 ccggatccat tatataccct ctagaccttt gatcttaccc cctttgatct tacccccttt
2701 gatcctgcag gcatgcaagc ttcagctgct cgagtaccgg atccattata taccctctag
2761 acctttgatc ttaccccctt tgatcttacc ccctttgatc ctgcaggcat gcaagcttca
2821 gctgctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggggatc
2881 cccgggtacc gagctccagg aacatccaaa ctgagcagcc ggggtccccc ccacccccca
2941 ccccgcccct cccggcaact ttgagcctgt gctgggacag agcctctagt tcctaaatta
3001 gtccatgagg tcagaggcag cactgccatt gtaacgcgat tggagaggat cacgtcaccg
3061 gacacgcccc caggcatctc cctgggtctc ctaaacttgg cggggagaag ttttagccct
3121 taagttttag cctttaaccc ccatattcag aactgtgcga gttggcgaaa ccccacaaat
3181 cacaacaaac tgtacacaac accgagctag aggtgatctt tcttgtccat tccacacagg
3241 ccttagtaat gcgtcgccat agcaacagtg tcactagtag caccagcact tccccacacc
3301 ctccccctca ggaatccgta ctctccagtg aaccccagaa acctctggag agttctggac
3361 aagggcggaa cccacaactc cgattactca agggaggcgg ggaagctcca ccagacgcga
3421 aactgctgga agattcctgg ccccaaggcc tcctccggct cgctgattgg cccagcggag
3481 agtgggcggg gccggtgaag actccttaaa ggcgcagggc ggcgagcagg tcaccagacg
3541 ctgacagcta ctcagaacca aatctggttc catccagaga caagcgaaga caagagaagc
3601 agagcgagcg gcgcgttccc gatcctcggc caggaccagc cttccccaga gcatccctgc
3661 cgcggagcgc aaccttccca ggagcatccc tgccgcggag cgcaactttc cccggagcat
3721 ccacgccgcg gagcgcagcc ttccagaagc acagctagcg ctaccggact cagatctcga
3781 gctcaagctt cgaattcgcc accatgccag agccagcgaa gtctgctccc gccccgaaaa
3841 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
3901 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
3961 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc
4021 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca
4081 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
4141 ccgagggtac taaggccgtc accaagtaca ccagcgctaa ggatccaccg gtcgccacca
4201 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
4261 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
4321 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
4381 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
4441 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
4501 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
4561 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
4621 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
4681 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
4741 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
4801 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
4861 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
4921 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
4981 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
5041 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
5101 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
5161 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
5221 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
5281 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
5341 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
5401 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
5461 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
5521 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
5581 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
5641 ggggaaatgt gcgcggaacc cctatttgtt tattttt
//
