| 质粒名称: | pCAG-DsRed |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11151 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGEN |
| 载体出品公司: |
Available at Addgene (#11160)
|
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4779 bp |
| 完整质粒大小: | -- |
| 载体修饰: | |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | DsRed2 from Discosoma sp,DsRed,red fluorescent protein |
| 物种来源: | -- |
| 插入大小: | 728 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI (未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | pCAG-F |
| 3' 测序引物 | DsRed-R |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-DsRed was a gift from Connie Cepko (Addgene plasmid # 11151 ; http://n2t.net/addgene:11151 ; RRID:Addgene_11151) |
| 参考文献部分: | Electroporation and RNA interference in the rodent retina in vivo and in vitro. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22. 10.1073/pnas.2235688100 PubMed 14603031 |
买家导航
pCAG-DsRed载体 pCAGDsRed质粒基本信息
pCAG-DsRed质粒图谱 pCAGDsRed载体图谱和pCAG-DsRed载体序列 pCAGDsRed质粒序列多克隆位点信息
pCAG-DsRed质粒 pCAGDsRed载体简介
pCAG-DsRed质粒序列 pCAGDsRed载体序列
LOCUS pCAG-DsRed 5507 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression vector for expression of DsRed.
ACCESSION .
VERSION .
KEYWORDS pCAG-DsRed
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5507)
AUTHORS Matsuda T, Cepko CL
TITLE Electroporation and RNA interference in the rodent retina in vivo
and in vitro.
JOURNAL Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
PUBMED 14603031
REFERENCE 2 (bases 1 to 5507)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5507
/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 1763..1772
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1769..2446
/codon_start=1
/product="improved tetrameric variant of DsRed fluorescent
protein"
/label=DsRed2
/note="mammalian codon-optimized"
/translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
LFL"
primer_bind complement(1949..1969)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 2361..2384
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(2531..2550)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 2596..2651
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2650..2669)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3010..3026)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3010..3026)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3023..3045)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3034..3050
/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(3058..3088)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3103..3124
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3183..3378
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3229..3364
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3291..3310
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 3384..3518
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3421..3440)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3475..3494
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3586..3603)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3757..4345)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3837..3856)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4516..5376)
/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 5139..5158
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5377..5481)
/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 tcgccaccat ggcctcctcc gagaacgtca tcaccgagtt
1801 catgcgcttc aaggtgcgca tggagggcac cgtgaacggc cacgagttcg agatcgaggg
1861 cgagggcgag ggccgcccct acgagggcca caacaccgtg aagctgaagg tgaccaaggg
1921 cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt
1981 gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt
2041 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gcgaccgtga cccaggactc
2101 ctccctgcag gacggctgct tcatctacaa ggtgaagttc atcggcgtga acttcccctc
2161 cgacggcccc gtgatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
2221 cccccgcgac ggcgtgctga agggcgagac ccacaaggcc ctgaagctga aggacggcgg
2281 ccactacctg gtggagttca agtccatcta catggccaag aagcccgtgc agctgcccgg
2341 ctactactac gtggacgcca agctggacat cacctcccac aacgaggact acaccatcgt
2401 ggagcagtac gagcgcaccg agggccgcca ccacctgttc ctgtagcggc cgcactcctc
2461 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
2521 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
2581 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
2641 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
2701 atttggttta gagtttggca acatatgcca tatgctggct gccatgaaca aaggtggcta
2761 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
2821 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
2881 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
2941 ccagtcatag ctgtccctct tctcttatga agatccctcg acctgcagcc caagcttggc
3001 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
3061 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
3121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
3181 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
3241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
3301 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
3361 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
3421 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
3481 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
3541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
3601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
3661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
3721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
3781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
3841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
3901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
3961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
4021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
4081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
4141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
4201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
4261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
4321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
4381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
4441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
4501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
4561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
4621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
4681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
4741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
4801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
4861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
4921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
4981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
5041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
5101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
5161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
5221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
5281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
5341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
5401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
5461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
