| 质粒名称: | pcDNA3.1-mCherry |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 128744 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 5300 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 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: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1-mCherry was a gift from David Bartel (Addgene plasmid # 128744 ; http://n2t.net/addgene:128744 ; RRID:Addgene_128744) |
| 参考文献部分: | A Network of Noncoding Regulatory RNAs Acts in the Mammalian Brain. Kleaveland B, Shi CY, Stefano J, Bartel DP. Cell. 2018 Jul 12;174(2):350-362.e17. doi: 10.1016/j.cell.2018.05.022. Epub 2018 Jun 7. 10.1016/j.cell.2018.05.022 PubMed 29887379 |
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 pcDNA3.1-mCherry 6462 bp ds-DNA circular SYN 14-AUG-2019
DEFINITION Expresses mCherry in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6462)
AUTHORS Kleaveland B, Shi CY, Stefano J, Bartel DP
TITLE A Network of Noncoding Regulatory RNAs Acts in the Mammalian Brain.
JOURNAL Cell. 2018 Jul 12;174(2):350-362.e17. doi:
10.1016/j.cell.2018.05.022. Epub 2018 Jun 7.
PUBMED 29887379
REFERENCE 2 (bases 1 to 6462)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 14, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6462
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 724..733
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 730..1440
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(875..893)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1143..1162
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1819..1836)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1825..2049
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2095..2523
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2182..2201)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2392..2413
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2532..2552)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2537..2866
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2717..2852
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2779..2798
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2933..3727
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2987..3006)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3597..3616
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3901..4022
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3938..3957)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3992..4011
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4071..4087)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4071..4087)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4084..4106)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4095..4111
/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(4119..4149)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4164..4185
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4302..4319)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4473..5061)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4553..4572)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5232..6092)
/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 5855..5874
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6093..6197)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6272..6291)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
721 ttcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
781 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
841 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
901 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
961 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
1021 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
1081 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
1141 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
1201 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
1261 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
1321 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
1381 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtga
1441 gcggccgctc gagtctagag ggccgggtaa gctcgctttc ttgctgtcca atttctatta
1501 aaggttcctt tgttccctaa gtccaactac taaactgggg attcctgggc cctgaagaag
1561 ggcccctcga ctaagtccaa ctactaaact gggccctgaa gaagggccca tatagggccc
1621 tgaagaaggg ccctatcgag gatattatct cgactaagtc caactactaa actgggccct
1681 gaagaagggc ccatataggg ccctgaagaa gggccctatc gaggatatta tctcgaggat
1741 attatgaagg gccttgagca tctggattct attctatagt gtcacctgct agagggcccg
1801 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
1861 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
1921 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
1981 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
2041 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
2101 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
2161 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
2221 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
2281 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
2341 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
2401 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
2461 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
2521 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
2581 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
2641 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
2701 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
2761 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
2821 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
2881 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
2941 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
3001 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
3061 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
3121 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
3181 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
3241 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
3301 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
3361 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
3421 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
3481 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
3541 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
3601 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
3661 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
3721 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
3781 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
3841 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
3901 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
3961 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
4021 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
4081 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
4141 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
4201 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
4261 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
4321 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
4381 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
4441 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
4501 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
4561 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
4621 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
4681 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
4741 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
4801 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
4861 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
4921 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
4981 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
5041 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
5101 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
5161 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
5221 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
5281 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
5341 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
5401 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
5461 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
5521 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
5581 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
5641 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
5701 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
5761 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
5821 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
5881 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
5941 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
6001 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
6061 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
6121 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
6181 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
6241 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
6301 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
6361 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
6421 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
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