| 质粒名称: | HBx |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 65463 |
| 存储实验室: | Wang-Shick Ryu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 完整质粒大小: | 6763bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 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: | HBV X protein / HBx |
| 物种来源: | Hepatitis B virus |
| 插入大小: | 462 bp |
| 标签/融合蛋白: | Flag (C terminal on insert) |
|
文献引用方法 |
|
| 材料和方法部分: | HBx was a gift from Wang-Shick Ryu (Addgene plasmid # 65463 ; http://n2t.net/addgene:65463 ; RRID:Addgene_65463) |
| 参考文献部分: | Hepatitis B virus X protein is essential for the activation of Wnt/beta-catenin signaling in hepatoma cells. Cha MY, Kim CM, Park YM, Ryu WS. Hepatology. 2004 Jun;39(6):1683-93. 10.1002/hep.20245 PubMed 15185310 |
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 HBx 6763 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION expresses HBx in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS HBx
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6763)
AUTHORS Cha MY, Kim CM, Park YM, Ryu WS
TITLE Hepatitis B virus X protein is essential for the activation of
Wnt/beta-catenin signaling in hepatoma cells.
JOURNAL Hepatology. 2004 Jun;39(6):1683-93.
PUBMED 15185310
REFERENCE 2 (bases 1 to 6763)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6763
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 481..546
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
polyA_signal 557..1179
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(756..775)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(1203..1223)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1221..1518
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind 1450..1469
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 1572..2366
/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(1626..1645)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2236..2255
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3018..3152
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3055..3074)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3109..3128
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3157..3176)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3158..3176)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3190..3206)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3190..3206)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3203..3225)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3214..3230
/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(3238..3268)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3283..3304
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3421..3438)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3592..4180)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3672..3691)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4351..5211)
/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 4974..4993
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5212..5316)
/gene="bla"
/label=AmpR promoter
rep_origin 5343..5798
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5430..5449)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5640..5661
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5924..5946
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5938..5955
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5939..5955
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 6116..6495
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 6496..6699
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 6650..6670
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
ORIGIN
1 atggctgcta ggctgtgctg ccaactggat cctgcgcggg acgtcctttg tttacgtccc
61 gtcggcgctg aatcctgcgg acgacccttc tcggggtcgc ttgggactct ctcgtcccct
121 tctccgtctg ccgttccgac cgaccacggg gcgcacctct ctttacgcgg actccccgtc
181 tgtgccttct catctgccgg accgtgtgca cttcgcttca cctctgcacg tcgcatggag
241 accaccgtga acgcccacca aatattgccc aaggtcttac ataagaggac tcttggactc
301 tcagcaatgt caacgaccga ccttgaggca tacttcaaag actgtttgtt taaagactgg
361 gaggagttgg gggaggagat taggttaaag gtctttgtac taggaggctg taggcataaa
421 ttggtctgcg caccagcacc atgcaacttt ttcacctctg cctctagagg atcccgggct
481 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
541 gacaagtagt gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
601 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
661 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
721 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
781 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
841 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
901 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
961 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
1021 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
1081 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
1141 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
1201 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
1261 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
1321 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
1381 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
1441 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
1501 gtgaggaggc ttttttggag gaattgatca gcttgggatc tgatcaagag acaggatgag
1561 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
1621 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
1681 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
1741 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
1801 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
1861 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
1921 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
1981 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
2041 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
2101 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
2161 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
2221 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
2281 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
2341 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
2401 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
2461 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
2521 ggagttcttc gcccaccccg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag
2581 gctggacgac ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag
2641 cagcggctat ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc
2701 tttggtcgac ccggacggga cgctcctgcg cctgatacag aacgaattgc ttgcaggcat
2761 ctcatgagtg tgtcttcccg ttttccgcct gaggtcactg cgtggatgga gcgctggcgc
2821 ctgctgcgcg acggcgagct gctcaccacc cactcgccaa gctggaaccg taaaaaggcc
2881 gcgttgctgg cgtttttcca taggctccgc cgatcataat cagccatacc acatttgtag
2941 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
3001 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
3061 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
3121 aactcatcaa tgtatcttat catgtctgga tcaattccct atagtgagtc gtattaaatt
3181 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
3241 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
3301 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
3361 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
3421 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
3481 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
3541 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
3601 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
3661 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
3721 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
3781 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
3841 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
3901 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
3961 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
4021 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
4081 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
4141 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
4201 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
4261 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
4321 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
4381 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
4441 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
4501 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
4561 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
4621 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
4681 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
4741 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
4801 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
4861 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
4921 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
4981 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
5041 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
5101 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
5161 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
5221 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
5281 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
5341 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
5401 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
5461 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
5521 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
5581 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
5641 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
5701 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
5761 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc
5821 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
5881 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
5941 aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca attagccata
6001 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
6061 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
6121 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
6181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
6241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
6301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
6361 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
6421 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
6481 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
6541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
6601 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
6661 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagaat
6721 taagcttctc tgttgtccta tcccgcaaat atacatcgtt tcc
//
