| 质粒名称: | pCAG-GBP1-10gly-lexADBD |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50795 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAG-GFP |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4823 bp |
| 完整质粒大小: | 5831 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | GBP1-lexA DNA binding domain fusion protein,GBP1-10gly-lexADBD,DBDG |
| 物种来源: | Synthetic |
| 插入大小: | 1008 bp |
| 启动子 | CAG |
| 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 | GGACTTCCTTTGTCCCAAATCTG |
| 3' 测序引物 | TAGCCAGAAGTCAGATGCTC |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-GBP1-10gly-lexADBD was a gift from Connie Cepko (Addgene plasmid # 50795 ; http://n2t.net/addgene:50795 ; RRID:Addgene_50795) |
| 参考文献部分: | A nanobody-based system using fluorescent proteins as scaffolds for cell-specific gene manipulation. Tang JC, Szikra T, Kozorovitskiy Y, Teixiera M, Sabatini BL, Roska B, Cepko CL. Cell. 2013 Aug 15;154(4):928-39. doi: 10.1016/j.cell.2013.07.021. 10.1016/j.cell.2013.07.021 PubMed 23953120 |
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 pCAG-GBP1-10gly-lexADBD 5831 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of a GFP-dependent transcription factor component in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pCAG-GBP1-10gly-lexADBD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5831)
AUTHORS Tang JC, Szikra T, Kozorovitskiy Y, Teixiera M, Sabatini BL, Roska
B, Cepko CL
TITLE A nanobody-based system using fluorescent proteins as scaffolds for
cell-specific gene manipulation.
JOURNAL Cell. 2013 Aug 15;154(4):928-39. doi: 10.1016/j.cell.2013.07.021.
PUBMED 23953120
REFERENCE 2 (bases 1 to 5831)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5831
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1594..1616
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1678..1697
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1762..1771
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1777..2118
/codon_start=1
/product="GFP-binding fragment of a single-chain camelid
antibody (Rothbauer et al., 2008)"
/label=GFP nanobody
/translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYSNVNVGFEYW
GQGTQVTVSS"
CDS 2161..2769
/codon_start=1
/gene="lexA"
/product="E. coli DNA-binding protein that represses genes
involved in the SOS response to DNA damage"
/label=LexA
/note="The N-terminal DNA-binding domain recognizes the
16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
/translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
primer_bind 2704..2725
/label=LexA
/note="LexA DNA binding domain, forward primer"
primer_bind complement(2855..2874)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2920..2975
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2974..2993)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3334..3350)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3334..3350)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3347..3369)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3358..3374
/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(3382..3412)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3427..3448
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3507..3702
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3553..3688
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3615..3634
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3708..3842
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3745..3764)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3799..3818
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3910..3927)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4081..4669)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4161..4180)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4840..5700)
/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 5463..5482
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5701..5805)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
1741 gcgggcccgg gatccaccgg tgccaccatg gccgacgtgc agctcgtgga atccggagga
1801 gcactggtcc agcctggggg atcactgaga ctgtcttgtg ccgcttctgg ctttcctgtg
1861 aatcggtact ctatgagatg gtacaggcag gcacctggaa aagagcgaga atgggtcgcc
1921 ggaatgtcat ctgctggcga taggagttcc tacgaggata gcgtgaaagg acggttcacc
1981 attagtaggg acgatgctag aaacaccgtg tacctccaga tgaactccct gaaacctgag
2041 gacactgctg tgtactactc caacgtcaat gtgggctttg aatactgggg acagggcaca
2101 caagtgactg ttagttctgc tagcggagga ggtggtggag gtggaggtgg aggtactagt
2161 atgaaagcgt taacggccag gcaacaagag gtgtttgatc tcatccgtga tcacatcagc
2221 cagacaggta tgccgccgac gcgtgcggaa atcgcgcagc gtttggggtt ccgttcccca
2281 aacgcggctg aagaacatct gaaggcgctg gcacgcaaag gcgttattga aattgtttcc
2341 ggcgcatcac gcgggattcg tctgttgcag gaagaggaag aagggttgcc gctggtaggt
2401 cgtgtggctg ccggtgaacc acttctggcg caacagcata ttgaaggtca ttatcaggtc
2461 gatccttcct tattcaagcc gaatgctgat ttcctgctgc gcgtcagcgg gatgtcgatg
2521 aaagatatcg gcattatgga tggtgacttg ctggcagtgc ataaaactca ggatgtacgt
2581 aacggtcagg tcgttgtcgc acgtattgat gacgaagtta ccgttaagcg cctgaaaaaa
2641 cagggcaata aagtcgaact gttgccagaa aatagcgagt ttaaaccaat tgtcgttgac
2701 cttcgtcagc agagcttcac cattgaaggt ctggcggttg gggttattcg caacggcgac
2761 tggctgtaag cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
2821 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
2881 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
2941 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
3001 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gccatatgct
3061 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
3121 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
3181 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
3241 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
3301 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
3361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
3421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
3481 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
3541 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
3601 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
3661 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
3721 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
3781 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
3841 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
3901 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
3961 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
4021 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
4081 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
4141 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
4201 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
4261 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
4321 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
4381 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
4441 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
4501 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
4561 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
4621 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
4681 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
4741 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
4801 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
4861 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
4921 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
4981 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
5041 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
5101 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
5161 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
5221 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
5281 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
5341 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
5401 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
5461 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
5521 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
5581 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
5641 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
5701 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
5761 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
5821 agtgccacct g
//
