| 质粒名称: | phage UbiC G3BP1-SNAP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 119949 |
| 存储实验室: | Jeffrey Chao |
|
载体骨架基本信息 |
|
| 载体名称: | pHAGE-UbiC |
| 空载体大小: | 7000 bp |
| 完整质粒大小: | 8219 bp |
| 载体类型: | 哺乳动物表达、细菌表达、慢病毒 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | G3BP1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1398 bp |
| 启动子: | UbiC |
| 标签/融合蛋白: | SNAP（插入件上的C端子） |
| 5’测序引物 | GTGTGGCACAGCTAGTTCC |
|
文献引用方法 |
|
| 材料和方法部分: | phage UbiC G3BP1-SNAP was a gift from Jeffrey Chao (Addgene plasmid # 119949 ; http://n2t.net/addgene:119949 ; RRID:Addgene_119949) |
| 参考文献部分: | Single-Molecule Imaging of mRNA Localization and Regulation during the Integrated Stress Response. Wilbertz JH, Voigt F, Horvathova I, Roth G, Zhan Y, Chao JA. Mol Cell. 2019 Jan 4. pii: S1097-2765(18)31036-0. doi: 10.1016/j.molcel.2018.12.006. 10.1016/j.molcel.2018.12.006 PubMed 30661979 |
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 UbiC G3BP1-SNAP 8219 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Lentiviral vector expressing G3BP1-SNAP fusion protein.
ACCESSION .
VERSION .
KEYWORDS phage UbiC G3BP1-SNAP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8219)
AUTHORS Wilbertz JH, Voigt F, Horvathova I, Roth G, Zhan Y, Chao JA
TITLE Single-Molecule Imaging of mRNA Localization and Regulation during
the Integrated Stress Response.
JOURNAL Mol Cell. 2019 Jan 4. pii: S1097-2765(18)31036-0. doi:
10.1016/j.molcel.2018.12.006.
PUBMED 30661979
REFERENCE 2 (bases 1 to 8219)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8219
/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."
CDS 1721..1765
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
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"
regulatory 2625..2634
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4047..4589
/codon_start=1
/product="human O6-alkylguanine-DNA-alkyltransferase
variant that forms covalent bonds with benzylguanine
derivatives"
/label=SNAP-tag(R)
/note="fast-reacting variant, optimized for mammalian
expression"
/translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
KEWLLAHEGHRLGKPGLG"
misc_feature 4609..5197
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4662..4682)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5080..5091)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5272..5505
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5624..5642)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5710..5814
/gene="bla"
/label=AmpR promoter
CDS 5815..6675
/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(6033..6052)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6846..7434
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7335..7354
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7588..7605
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 7697..7716
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 7845..7866
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7881..7911
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7919..7935
/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 7924..7946
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7943..7959
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7943..7959
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 8009..8028
/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 cgccgccacc atggtgatgg
2641 agaagcctag tcccctgctg gtcgggcggg aatttgtgag acagtattac acactgctga
2701 accaggcccc agacatgctg catagatttt atggaaagaa ctcttcttat gtccatgggg
2761 gattggattc aaatggaaag ccagcagatg cagtctacgg acagaaagaa atccacagga
2821 aagtgatgtc acaaaacttc accaactgcc acaccaagat tcgccatgtt gatgctcatg
2881 ccacgctaaa tgatggtgtg gtagtccagg tgatggggct tctctctaac aacaaccagg
2941 ctttgaggag attcatgcaa acgtttgtcc ttgctcctga ggggtctgtt gcaaataaat
3001 tctatgttca caatgatatc ttcagatacc aagatgaggt ctttggtggg tttgtcactg
3061 agcctcagga ggagtctgaa gaagaagtag aggaacctga agaaagacag caaacacctg
3121 aggtggtacc tgatgattct ggaactttct atgatcaggc agttgtcagt aatgacatgg
3181 aagaacattt agaggagcct gttgctgaac cagagcctga tcctgaacca gaaccagaac
3241 aagaacctgt atctgaaatc caagaggaaa agcctgagcc agtattagaa gaaactgccc
3301 ctgaggatgc tcagaagagt tcttctccag cacctgcaga catagctcag acagtacagg
3361 aagacttgag gacattttct tgggcatctg tgaccagtaa gaatcttcca cccagtggag
3421 ctgttccagt tactgggata ccacctcatg ttgttaaagt accagcttca cagccccgtc
3481 cagagtctaa gcctgaatct cagattccac cacaaagacc tcagcgggat caaagagtgc
3541 gagaacaacg aataaatatt cctccccaaa ggggacccag accaatccgt gaggctggtg
3601 agcaaggtga cattgaaccc cgaagaatgg tgagacaccc tgacagtcac caactcttca
3661 ttggcaacct gcctcatgaa gtggacaaat cagagcttaa agatttcttt caaagttatg
3721 gaaacgtggt ggagttgcgc attaacagtg gtgggaaatt acccaatttt ggttttgttg
3781 tgtttgatga ttctgagcct gttcagaaag tccttagcaa caggcccatc atgttcagag
3841 gtgaggtccg tctgaatgtc gaagagaaga agactcgagc tgccagggaa ggcgaccgac
3901 gagataatcg ccttcgggga cctggaggcc ctcgaggtgg gctgggtggt ggaatgagag
3961 gccctccccg tggaggcatg gtgcagaaac caggatttgg agtgggaagg gggcttgcgc
4021 cacggcaggg gggggggggg tctagagaca aagactgcga aatgaagcgc accaccctgg
4081 atagccctct gggcaagctg gaactgtctg ggtgcgaaca gggcctgcac cgtatcatct
4141 tcctgggcaa aggaacatct gccgccgacg ccgtggaagt gcctgcccca gccgccgtgc
4201 tgggcggacc agagccactg atgcaggcca ccgcctggct caacgcctac tttcaccagc
4261 ctgaggccat cgaggagttc cctgtgccag ccctgcacca cccagtgttc cagcaggaga
4321 gctttacccg ccaggtgctg tggaaactgc tgaaagtggt gaagttcgga gaggtcatca
4381 gctacagcca cctggccgcc ctggccggca atcccgccgc caccgccgcc gtgaaaaccg
4441 ccctgagcgg aaatcccgtg cccattctga tcccctgcca ccgggtggtg cagggcgacc
4501 tggacgtggg gggctacgag ggcgggctcg ccgtgaaaga gtggctgctg gcccacgagg
4561 gccacagact gggcaagcct gggctgggtt aatgaatcga tagatcctaa tcaacctctg
4621 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
4681 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
4741 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
4801 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
4861 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
4921 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
4981 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
5041 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
5101 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
5161 cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta agaccaatga
5221 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
5281 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
5341 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
5401 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
5461 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
5521 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcccg
5581 ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt gatacgccta
5641 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
5701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
5761 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
5821 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
5881 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
5941 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
6001 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
6061 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
6121 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
6181 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
6241 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
6301 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
6361 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
6421 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
6481 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
6541 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
6601 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
6661 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
6721 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
6781 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
6841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
6901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
6961 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
7021 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
7081 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
7141 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
7201 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
7261 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
7321 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
7381 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
7441 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
7501 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
7561 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
7621 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag caagctcatg
7681 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
7741 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgtggcac
7801 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
7861 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
7921 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttcacaaat
7981 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
8041 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
8101 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
8161 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
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