| 质粒名称: | pLV-75-2 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 73036 |
| 存储实验室: | Pantelis Tsoulfas |
|
载体骨架基本信息 |
|
| 载体名称: | pRRLsinPPT.CMV.MCS.Wpre |
| 载体出品公司: | erNaldini |
| 空载体大小: | 6700 bp |
| 完整质粒大小: | 7600 bp |
| 载体类型: | 哺乳细胞表达、 Lentiviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
|
插入基因信息 |
|
| 插入基因名称: | p75-2 / NT-3/D15A/R113A/K114A |
| 物种来源: | Human |
| 插入大小: | 774bp |
| 基因突变: | Mature Human NT-3 gene exchanged Aspartic 15 to alanine, Arginine 113 to Alanine and Lysine 114 to Alanine |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XbaI and BamHI(未破坏) |
| 3’克隆位点: | none (destroyed during cloning) |
| 5’测序引物 | CMV_frw_primer |
|
文献引用方法 |
|
| 材料和方法部分: | pLV-75-2 was a gift from Pantelis Tsoulfas (Addgene plasmid # 73036 ; http://n2t.net/addgene:73036 ; RRID:Addgene_73036) |
| 参考文献部分: | A multifunctional neurotrophin with reduced affinity to p75NTR enhances transplanted Schwann cell survival and axon growth after spinal cord injury. Enomoto M, Bunge MB, Tsoulfas P. Exp Neurol. 2013 Oct;248:170-82. doi: 10.1016/j.expneurol.2013.06.013. Epub 2013 Jun 20. 10.1016/j.expneurol.2013.06.013 PubMed 23792206 |
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 pLV-75-2 7597 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express an NT-3-BDNF hybrid neurotrophic factor, with reduced
affinity to p75NTR, in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLV-75-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7597)
AUTHORS Enomoto M, Bunge MB, Tsoulfas P
TITLE A multifunctional neurotrophin with reduced affinity to p75NTR
enhances transplanted Schwann cell survival and axon growth after
spinal cord injury.
JOURNAL Exp Neurol. 2013 Oct;248:170-82. doi:
10.1016/j.expneurol.2013.06.013. Epub 2013 Jun 20.
PUBMED 23792206
REFERENCE 2 (bases 1 to 7597)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7597
/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"
misc_feature 5654..6242
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5707..5727)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6125..6136)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6329..6562
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6634..6755
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6671..6690)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6725..6744
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6795..6930
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6857..6876
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(6950..6969)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6951..6969)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6979..6996)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6979..6995)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6988..7010)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7137..7592
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7224..7243)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7434..7455
/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 aagacaccga ctctagagga
4741 tcctacaggt gaacaagacc atgtccatct tgttttatgt gatatttctc gcttatctcc
4801 gtggcatcca aggtaacaac atggatcaaa ggagtttgcc agaagactcg ctcaattccc
4861 tcattattaa gctgatccag gcagatattt tgaaaaacaa gctctccaag cagatggtgg
4921 acgttaagga aaattaccag agcaccctgc ccaaagctga ggctccccga gagccggagc
4981 ggggagggcc cgccaagtca gcattccagc cggtgattgc aatggacacc gaactgctgc
5041 gacaacagag acgctacaac tcaccgcggg tcctgctgag cgacagcacc cccttggagc
5101 ccccgccctt gtatctcatg gaggattacg tgggcagccc cgtggtggcg aacagaacat
5161 cacggcggaa acggtacgcg gagcataaga gtcaccgagg ggagtactcg gtatgtgcca
5221 gtgaaagctt gtgggtgacc gacaagtcat cggccatcga cattcgggga caccaggtca
5281 cggtgctggg ggagatcaaa acgggcaact ctcccgtcaa acaatatttt tatgaaacgc
5341 gatgtaagga agccaggccg gtcaaaaacg gttgcagggg tattgatgat aaacactgga
5401 actctcagtg caaaacatcc caaacctacg tccgagcact gacttcagag aacaataaac
5461 tcgtgggctg gcggtggata cggatagaca cgtcctgtgt gtgtgccttg tctgcagcaa
5521 tcggaagaac atgaattggc atctctcccc atatataaat tattacttta aattatatga
5581 tatgcatgta gcatataaat gtttatattg tttttatata ttataagtta tcaagcttat
5641 cgataccgtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
5701 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
5761 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
5821 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
5881 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
5941 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
6001 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
6061 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
6121 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
6181 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
6241 gcctggaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
6301 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
6361 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
6421 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
6481 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
6541 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
6601 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
6661 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
6721 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
6781 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
6841 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
6901 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
6961 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
7021 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
7081 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
7141 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
7201 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
7261 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
7321 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
7381 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
7441 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
7501 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
7561 gaattttaac aaaatattaa cgcttacaat ttaggtg
//
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