| 质粒名称: | pcDNA3.1-Flag HBx |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 42596 |
| 存储实验室: | Ding Xue |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 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: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Hepatitis B virus protein X,HBx |
| 物种来源: | Hepatitis B virus |
| 插入大小: | 492 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | Flag（插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | EcoRV (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1-Flag HBx was a gift from Ding Xue (Addgene plasmid # 42596 ; http://n2t.net/addgene:42596 ; RRID:Addgene_42596) |
| 参考文献部分: | Hepatitis B virus X protein targets Bcl-2 proteins to increase intracellular calcium, required for virus replication and cell death induction. Geng X, Huang C, Qin Y, McCombs JE, Yuan Q, Harry BL, Palmer AE, Xia NS, Xue D. Proc Natl Acad Sci U S A. 2012 Nov 6;109(45):18471-6. doi: 10.1073/pnas.1204668109. Epub 2012 Oct 22. 10.1073/pnas.1204668109 PubMed 23091012 |
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 pcDNA3.1-Flag HBx 5859 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1-Flag HBx
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5859)
AUTHORS Geng X, Huang C, Qin Y, McCombs JE, Yuan Q, Harry BL, Palmer AE, Xia
NS, Xue D
TITLE Hepatitis B virus X protein targets Bcl-2 proteins to increase
intracellular calcium, required for virus replication and cell death
induction.
JOURNAL Proc Natl Acad Sci U S A. 2012 Nov 6;109(45):18471-6. doi:
10.1073/pnas.1204668109. Epub 2012 Oct 22.
PUBMED 23091012
REFERENCE 2 (bases 1 to 5859)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5859
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 904..927
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(1453..1470)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1459..1683
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1729..2157
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1816..1835)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2026..2047
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2166..2186)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2171..2500
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2351..2486
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2413..2432
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2567..3361
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2621..2640)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3231..3250
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3535..3656
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3572..3591)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3626..3645
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3705..3721)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3705..3721)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3718..3740)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3729..3745
/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)."
promoter complement(3753..3783)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3798..3819
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3936..3953)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4107..4692)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4187..4206)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4863..5723)
/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 5486..5505
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5724..5828)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 atggactaca aggatgacga tgacaagatg gctgctaggc tgtactgcca actggatcct
961 tcgcgggacg tcctttgttt acgtcccgtc ggcgctgaat cccgcggacg acccctctcg
1021 gggccgcttg ggactctctc gtccccttct ccgtctgccg ttccagccga ccacggggcg
1081 cacctctctt tacgcggtct ccccgtctgt gccttctcat ctgccggtcc gtgtgcactt
1141 cgcttcacct ctgcacgttg catggagacc accgtgaacg cccatcagat cctgcccaag
1201 gtcttacata agaggactct tggactccca gcaatgtcaa cgaccgacct tgaggcctac
1261 ttcaaggact gtgtgtttaa ggactgggag gagctggggg aggagattag gttaaaggtc
1321 tttgtattag gaggctgtag gcataaattg gtctgcgcac cagcaccatg caactttttc
1381 acctctgcct aagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa
1441 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
1501 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
1561 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
1621 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
1681 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
1741 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
1801 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
1861 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
1921 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
1981 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
2041 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
2101 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
2161 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
2221 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
2281 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
2341 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
2401 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
2461 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
2521 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
2581 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
2641 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
2701 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
2761 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
2821 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
2881 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
2941 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
3001 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
3061 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
3121 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
3181 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
3241 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
3301 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
3361 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
3421 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
3481 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
3541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
3601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
3661 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
3721 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
3781 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
3841 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
3901 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
3961 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
4021 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
4081 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
4141 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
4201 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
4261 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
4321 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
4381 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
4441 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
4501 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
4561 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
4621 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
4681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
4741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
4801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
4921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
4981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
5041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
5101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
5161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
5221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
5281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
5341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
5401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
5461 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
5521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
5581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
5641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
5701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
5761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
5821 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
