| 质粒名称: | pGFP-HBx |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 24931 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C2 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 4700 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | HBx |
| 物种来源: | -- |
| 插入大小: | 529 bp |
| 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(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pGFP-HBx was a gift from Xin Wang (Addgene plasmid # 24931 ; http://n2t.net/addgene:24931 ; RRID:Addgene_24931) |
| 参考文献部分: | Interaction of the hepatitis B virus X protein with the Crm1-dependent nuclear export pathway. Forgues M, Marrogi AJ, Spillare EA, Wu CG, Yang Q, Yoshida M, Wang XW. J Biol Chem. 2001 Jun 22. 276(25):22797-803. 10.1074/jbc.M101259200 PubMed 11287420 |
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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 pGFP-HBx 5227 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pGFP-HBx
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5227)
AUTHORS Forgues M, Marrogi AJ, Spillare EA, Wu CG, Yang Q, Yoshida M, Wang
XW
TITLE Interaction of the hepatitis B virus X protein with the
Crm1-dependent nuclear export pathway.
JOURNAL J Biol Chem. 2001 Jun 22. 276(25):22797-803.
PUBMED 11287420
REFERENCE 2 (bases 1 to 5227)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5227
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 16..319
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 320..523
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 474..494
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 562..571
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 568..1284
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(613..634)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(874..893)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1221..1242
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1804..1830
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 1970..2091
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2007..2026)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2061..2080
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2098..2553)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2235..2256)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2447..2466
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2580..2684
/gene="bla"
/label=AmpR promoter
promoter 2686..3043
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2709..2729)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2894..3029
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2956..2975
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3078..3872
/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(3132..3151)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3742..3761
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4060..4079)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4104..4151
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4480..5068
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4969..4988
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
541 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
601 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
661 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
721 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
781 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
841 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
901 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
961 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
1021 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
1081 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
1141 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
1201 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
1261 ctcggcatgg acgagctgta caagtccggc cggactcaga tctcgagctc aagcttggta
1321 ccgagctcgg atctgatggc tgctcgggtg tgctgccaac tggatcctgc gcgggacgtc
1381 ctttgtctac gtcccgtcgg cgctgaatcc cgcggacgac ccgtctcggg gccgtttggg
1441 cctctaccgt cccttgcttt ctctgccgtt ccagccgacc acggggcgca cctctcttta
1501 cgcggtctcc ccgtctgtgc cttctcatct gccggaccgt gtgcacttcg cttcacctct
1561 gcacgtcgca tggagaccac cgtgaacggc caccaggtct tgcccaagct cttacataag
1621 aggactcttg gactctcagc aatgtcaaca accgaccttg aggcatactt caaagactgt
1681 ttgtttaaag actgggagga gttgggggag gagattaggt taaaggtctt tgtactagga
1741 ggctgtaggc ataaattggt ctgttcacca gcaccatgca actttttcac ctctgcctct
1801 agataccctt atgatgtgcc agattatgcc tgagggcccg ggatccaccg gatctagata
1861 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
1921 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
1981 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
2041 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
2101 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
2161 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
2221 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
2281 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
2341 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
2401 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
2461 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
2521 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
2581 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
2641 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
2701 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
2761 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
2821 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
2881 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
2941 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
3001 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
3061 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
3121 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
3181 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
3241 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
3301 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
3361 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
3421 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
3481 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
3541 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
3601 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
3661 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
3721 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
3781 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
3841 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
3901 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
3961 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
4021 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
4081 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
4141 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
4201 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
4261 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
4321 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
4381 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
4441 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
4501 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
4561 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
4621 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
4681 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
4741 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
4801 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
4861 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
4921 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
4981 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
5041 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
5101 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
5161 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
5221 atagccc
//
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