| 质粒名称: | pCAG-GBP6-10gly-VPminx4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50796 |
| 存储实验室: | Connie Cepko |
|
载体骨架基本信息 |
|
| 载体名称: | pCAG-GFP |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4823 bp |
| 完整质粒大小: | 5420 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: | GBP6-VPminx4 activation domain fusion protein,ADG,GBP6-10gly-VPminx4 |
| 物种来源: | Synthetic |
| 插入大小: | 597 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-GBP6-10gly-VPminx4 was a gift from Connie Cepko (Addgene plasmid # 50796 ; http://n2t.net/addgene:50796 ; RRID:Addgene_50796) |
| 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-GBP6-10gly-VPminx4 5419 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of a GFP-dependent transcription factor component in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pCAG-GBP6-10gly-VPminx4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5419)
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 5419)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5419
/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"
gap 1768..2150
/estimated_length=383
primer_bind complement(2443..2462)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2508..2563
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2562..2581)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2922..2938)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2922..2938)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2935..2957)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2946..2962
/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(2970..3000)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3015..3036
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3095..3290
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3141..3276
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3203..3222
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3296..3430
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3333..3352)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3387..3406
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3498..3515)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3669..4257)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3749..3768)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4428..5288)
/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 5051..5070
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5289..5393)
/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 tgccaccnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
1801 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
1861 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
1921 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
1981 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
2041 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
2101 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn gctagcggag
2161 gaggtggtgg aggtggaggt ggaggtcaat tgcctaccga cgccctggac gacttcgacc
2221 tggacatgct gcccgccgat gccctggatg attttgatct ggatatgctc cctgctgacg
2281 ctctcgatga cttcgatctc gatatgctgc cagccgacgc gctagacgat ttcgatctgg
2341 acatgttgcc aggctaagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt
2401 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
2461 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
2521 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
2581 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
2641 catatgctgg ctgccatgaa caaaggtggc tataaagagg tcatcagtat atgaaacagc
2701 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
2761 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
2821 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
2881 gaagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg
2941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
3001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
3061 ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag
3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt
3301 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
3361 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
3421 tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
3481 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
3541 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
3601 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
3661 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
3721 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
3781 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
3841 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
3901 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
3961 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
4021 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
4081 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
4141 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
4201 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
4261 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
4321 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
4381 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
4441 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
4501 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
4561 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
4621 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
4681 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
4741 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
4801 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
4861 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
4921 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
4981 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
5041 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
5101 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
5161 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
5221 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
5281 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
5341 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
5401 ccccgaaaag tgccacctg
//
