| 质粒名称: | phage-ubc-nls-ha-tdMCP-gfp |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 40649 |
| 存储实验室: | Robert Singer |
|
载体骨架基本信息 |
|
| 载体名称: | pHAGE-UBC-RIG (modified) |
| 载体出品公司: | see associated publication |
| 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: | 哺乳动物表达，慢病毒 |
| 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: | tdMCP-gfp,tandem dimer MS2 coat protein (MCP) |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | human ubiquitin C (UBC) |
| 标签/融合蛋白: |
NLS（主干上的N终端）
HA（主干上的N终端）
EGFP（主干上的C端）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | FUGW (5'-ATTACAGGGACAGCAGAGATCC-3') |
| 3' 测序引物 | WPRE-R (5'CATAGCGTAAAAGGAGCAACA-3') |
|
文献引用方法 |
|
| 材料和方法部分: | phage-ubc-nls-ha-tdMCP-gfp was a gift from Robert Singer (Addgene plasmid # 40649 ; http://n2t.net/addgene:40649 ; RRID:Addgene_40649) |
| 参考文献部分: | Fluorescence fluctuation spectroscopy enables quantitative imaging of single mRNAs in living cells. Wu B, Chao JA, Singer RH. Biophys J. 2012 Jun 20;102(12):2936-44. Epub 2012 Jun 19. 10.1016/j.bpj.2012.05.017 PubMed 22735544 |
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 phage-ubc-nls-ha-tdMCP-gfp 7772 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS phage-ubc-nls-ha-tdMCP-gfp
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7772)
AUTHORS Wu B, Chao JA, Singer RH
TITLE Fluorescence fluctuation spectroscopy enables quantitative imaging
of single mRNAs in living cells.
JOURNAL Biophys J. 2012 Jun 20;102(12):2936-44. Epub 2012 Jun 19.
PUBMED 22735544
REFERENCE 2 (bases 1 to 7772)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7772
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2058..2175
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2218..2616
/label=UbC promoter
/note="human ubiquitin C promoter"
CDS 2631..2651
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2655..2681
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2682..2693
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 3429..4148
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3474..3495)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3735..3754)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4082..4103
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 4162..4750
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4215..4235)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4633..4644)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4825..5058
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5177..5195)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5263..5367
/gene="bla"
/label=AmpR promoter
CDS 5368..6228
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5586..5605)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6399..6987
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6888..6907
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7141..7158
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 7250..7269
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 7398..7419
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7434..7464
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7472..7488
/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)."
primer_bind 7477..7499
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7496..7512
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7496..7512
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7562..7581
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtggc
2221 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
2281 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
2341 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
2401 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
2461 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgcaatgggc ccaaaaaaga
2641 aaagaaaagt tggctacccc tacgacgtgc ccgactacgc catcgaaggc cgccatatgc
2701 tagccgttaa aatggcttct aactttactc agttcgttct cgtcgacaat ggcggaactg
2761 gcgacgtgac tgtcgcccca agcaacttcg ctaacgggat cgctgaatgg atcagctcta
2821 actcgcgttc acaggcttac aaagtaacct gtagcgttcg tcagagctct gcgcagaatc
2881 gcaaatacac catcaaagtc gaggtgccta aaggcgcctg gcgttcgtac ttaaatatgg
2941 aactaaccat tccaattttc gccacgaatt ccgactgcga gcttattgtt aaggcaatgc
3001 aaggtctcct aaaagatgga aacccgattc cctcagcaat cgcagcaaac tccggcatct
3061 acgccatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga actggcgacg
3121 tgactgtcgc cccaagcaac ttcgctaacg ggatcgctga atggatcagc tctaactcgc
3181 gttcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag aatcgcaaat
3241 acaccatcaa agtcgaggtg cctaaaggcg cctggcgttc gtacttaaat atggaactaa
3301 ccattccaat tttcgccacg aattccgact gcgagcttat tgttaaggca atgcaaggtc
3361 tcctaaaaga tggaaacccg attccctcag caatcgcagc aaactccggc atctacgcgg
3421 attctagaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
3481 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
3541 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
3601 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
3661 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
3721 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
3781 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
3841 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
3901 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
3961 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
4021 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
4081 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
4141 acaagtaaat cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact
4201 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
4261 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
4321 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
4381 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
4441 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
4501 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
4561 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
4621 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
4681 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
4741 gcctccccgc ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
4801 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
4861 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
4921 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
4981 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
5041 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
5101 caaagaaatg aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat
5161 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
5221 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
5281 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
5341 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
5401 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
5461 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
5521 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
5581 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
5641 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
5701 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
5761 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
5821 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
5881 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
5941 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
6001 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
6061 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
6121 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
6181 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
6241 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
6301 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
6361 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
6421 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
6481 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
6541 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
6601 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
6661 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
6721 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
6781 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
6841 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
6901 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
6961 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
7021 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
7081 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
7141 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
7201 cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag
7261 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
7321 gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc
7381 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
7441 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
7501 aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt
7561 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
7621 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
7681 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
7741 atttgttaaa tatgtactac aaacttagta gt
//
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