| 质粒名称: | H2B-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11680 |
| 存储实验室: | Geoff Wahl |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4733bp |
| 完整质粒大小: | 5107 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin |
| 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: | Human histone H2B |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 380bp |
| GenBank编号 | X00088 |
| 启动子: | CMV |
| 标签/融合蛋白: | EGFP (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | KpnI (not destroyed) |
| 3’克隆位点: | BamHI (not destroyed) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | H2B-GFP was a gift from Geoff Wahl (Addgene plasmid # 11680 ; http://n2t.net/addgene:11680 ; RRID:Addgene_11680) |
| 参考文献部分: |
Histone-GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells. Kanda T, Sullivan KF, Wahl GM. Curr Biol. 1998 Mar 26. 8(7):377-85. 10.1016/S0960-9822(98)70156-3 PubMed 9545195
|
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载体基本信息
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载体简介
载体序列
LOCUS H2B-GFP 5107 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5107)
AUTHORS Kanda T, Sullivan KF, Wahl GM
TITLE Histone-GFP fusion protein enables sensitive analysis of chromosome
dynamics in living mammalian cells.
JOURNAL Curr Biol. 1998 Mar 26. 8(7):377-85.
PUBMED 9545195
REFERENCE 2 (bases 1 to 5107)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Curr Biol.
1998 Mar 26. 8(7):377-85."
FEATURES Location/Qualifiers
source 1..5107
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 146..337
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind 294..314
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(339..443)
/gene="bla"
/label=AmpR promoter
rep_origin 470..925
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(557..576)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 767..788
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 932..1053
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(943..962)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 997..1016
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(1176..1895)
/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 complement(1179..1895)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1221..1242)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1570..1589
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1829..1850
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 1892..1901
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS complement(1914..2291)
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAITKYTSAK"
promoter complement(2380..2583)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2409..2429)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 2584..2887
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(3062..3650)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3142..3161)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 3979..4026
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4051..4070
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(4258..5052)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS complement(4258..5052)
/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(4369..4388)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4979..4998
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(join(5087..5107,1..337))
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin join(5101..5107,1..129)
/label=SV40 ori
/note="SV40 origin of replication"
ORIGIN
1 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
61 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
121 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
181 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
241 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
301 ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat
361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
481 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
541 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
601 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
661 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
721 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
781 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
841 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
901 aacaaaatat taacgcttac aatttacgcc ttaagataca ttgatgagtt tggacaaacc
961 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
1021 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
1081 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
1141 ggtatggctg attatgatct agagtcgcgg ccgctttact tgtacagctc gtccatgccg
1201 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
1261 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
1321 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
1381 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
1441 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
1501 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
1561 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
1621 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
1681 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg
1741 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
1801 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
1861 accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtgg atccttagcg
1921 ctggtgtact tggtgatggc cttagtaccc tcggacacgg cgtgcttggc caactcccca
1981 ggcagcagca ggcgcacggc cgtctggatc tccctggagg tgatggtcga gcgcttgttg
2041 taatgcgcca ggcgggaagc ctcacctgcg atgcgctcga aaatgtcgtt cacaaacgaa
2101 ttcatgatgc ccatggcctt ggacgaaatg ccggtgtcag ggtggacctg cttcagaacc
2161 ttgtacacat agatggaata gctctccttg cggctgcgct tgcgcttctt gccgcctttc
2221 ttctgcgcct tagtcaccgc cttcttggag ccctttttcg gggcgggagc agacttcgct
2281 ggctctggca tggtggcggt accgtcgact gcagaattcg aagcttgagc tcgagatctg
2341 agtccggtag cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc
2401 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
2461 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
2521 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
2581 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
2641 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
2701 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
2761 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
2821 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
2881 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
2941 ataactaatg catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
3001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
3061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
3121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
3181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
3241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
3301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
3361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
3421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
3481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
3541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
3601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
3661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
3721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
3781 aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt
3841 tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa
3901 cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
3961 accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta
4021 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
4081 ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
4141 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
4201 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
4261 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
4321 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
4381 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
4441 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
4501 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
4561 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
4621 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
4681 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
4741 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
4801 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
4861 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
4921 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
4981 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
5041 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta
5101 ggcctcc
//
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