| 质粒名称: | pLV-mitoDsRed |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 44386 |
| 存储实验室: | Pantelis Tsoulfas |
|
载体骨架基本信息 |
|
| 载体名称: | pLV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6727 bp |
| 完整质粒大小: | 7600 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,慢病毒 |
| 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: | mitoDsRed |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 880 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: |
P1亚型F1F0-ATP合酶的61个aa靶向序列（插入物上的N末端）
DsRed（插入件上的C端子
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SpeI（克隆过程中销毁） |
| 3' 克隆位点: | EcoRV（克隆过程中销毁） |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | WPRE-R |
|
文献引用方法 |
|
| 材料和方法部分: | pLV-mitoDsRed was a gift from Pantelis Tsoulfas (Addgene plasmid # 44386 ; http://n2t.net/addgene:44386 ; RRID:Addgene_44386) |
| 参考文献部分: | Mislocalization of neuronal mitochondria reveals regulation of Wallerian degeneration and NMNAT/WLDS-mediated axon protection independent of axonal mitochondria. Kitay BM, McCormack R, Wang Y, Tsoulfas P, Zhai RG. Hum Mol Genet. 2013 Jan 24. 10.1093/hmg/ddt009 PubMed 23314018 |
买家导航
pLVmitoDsRed载体基本信息
pLVmitoDsRed载体图谱质粒图谱和多克隆位点信息
pLVmitoDsRed载体简介
pLVmitoDsRed载体序列
LOCUS pLV-mitoDsRed 7824 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION 3rd generation lentiviral vector.
ACCESSION .
VERSION .
KEYWORDS pLV-mitoDsRed
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7824)
AUTHORS Kitay BM, McCormack R, Wang Y, Tsoulfas P, Zhai RG
TITLE Mislocalization of neuronal mitochondria reveals regulation of
Wallerian degeneration and NMNAT/WLDS-mediated axon protection
independent of axonal mitochondria.
JOURNAL Hum Mol Genet. 2013 Jan 24.
PUBMED 23314018
REFERENCE 2 (bases 1 to 7824)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7824
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 21..125
/gene="bla"
/label=AmpR promoter
CDS 126..986
/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 complement(344..363)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1157..1745
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1646..1665
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1899..1916
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2033..2054
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2069..2099
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2107..2123
/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 2112..2134
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2131..2147
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2131..2147
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2166..2186
/label=T3
/note="T3 promoter, forward primer"
promoter 2168..2186
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2214..2440
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 2441..2621
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2668..2793
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3286..3519
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 3704..3748
/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 4015..4131
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 4154..4457
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4458..4661
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4612..4632
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 4658..4682
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 5069..5078
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 5075..5755
/codon_start=1
/product="wild-type DsRed"
/label=DsRed1
/note="mammalian codon-optimized"
/translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
HLFL"
primer_bind complement(5258..5278)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 5476..5495
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 5670..5693
/label=DsRed1-C
/note="DsRed1, forward primer"
misc_feature 5881..6469
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5934..5954)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6352..6363)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6556..6789
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6861..6982
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6898..6917)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6952..6971
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 7022..7157
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7084..7103
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(7177..7196)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(7178..7196)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(7206..7223)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7206..7222)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7215..7237)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7364..7819
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7451..7470)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7661..7682
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
2161 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
2221 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
2281 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
2341 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
2401 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
2461 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
2521 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
2581 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
2641 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
2701 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
2761 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
2821 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
2881 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
2941 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
3001 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
3061 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
3121 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
3181 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
3241 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
3301 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
3361 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
3421 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
3481 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
3541 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
3601 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
3661 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
3721 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
3781 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
3841 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
3901 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
3961 agacagagac agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa
4021 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
4081 agacatacaa actaaagaat tacaaaaaca aattacaaaa ttcaaaattt tatcgataag
4141 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
4201 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
4261 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
4321 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
4381 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
4441 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
4501 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
4561 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
4621 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
4681 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgc ccgggcgtac
4741 gggatcccac cggtgctcga gatgcatact agcgctaccg gactcagatc tcgagctcaa
4801 gcttgccacc atgcagaccg ccggggcatt attcatttct ccagctctga tccgctgttg
4861 taccaggggt ctaatcaggc ctgtgtctgc ctccttcttg aatagcccag tgaattcatc
4921 taaacagcct tcctacagca acttcccact ccaggtggcc agacgggagt tccagaccag
4981 tgttgtctcc cgggacattg acacagcagc catgttgaag cttcgaattc tgcagtcgac
5041 ggtaccgcgg gcccgggatc caccggtcgc caccatggtg cgctcctcca agaacgtcat
5101 caaggagttc atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc acgagttcga
5161 gatcgagggc gagggcgagg gccgccccta cgagggccac aacaccgtga agctgaaggt
5221 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg
5281 ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc
5341 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
5401 ccaggactcc tccctgcagg acggctgctt catctacaag gtgaagttca tcggcgtgaa
5461 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
5521 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc cacaaggccc tgaagctgaa
5581 ggacggcggc cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca
5641 gctgcccggc tactactacg tggactccaa gctggacatc acctcccaca acgaggacta
5701 caccatcgtg gagcagtacg agcgcaccga gggccgccac cacctgttcc tgtagcggcc
5761 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
5821 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgtttcgac
5881 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
5941 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
6001 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
6061 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
6121 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
6181 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
6241 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
6301 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
6361 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
6421 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
6481 gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
6541 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
6601 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
6661 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
6721 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
6781 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
6841 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
6901 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
6961 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
7021 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
7081 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
7141 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
7201 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
7261 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
7321 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
7381 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
7441 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
7501 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
7561 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
7621 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
7681 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
7741 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
7801 atattaacgc ttacaattta ggtg
//
