| 质粒名称: | SPLICS Mt-ER- Po Long P2A |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 164117 |
| 存储实验室: | Marisa Brini, Tito Calì |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Split GFP and YFP Mt-ER- Po Long P2A |
| 物种来源: | H. sapiens (human) |
| 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: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | SPLICS Mt-ER- Po Long P2A was a gift from Marisa Brini & Tito Calì (Addgene plasmid # 164117 ; http://n2t.net/addgene:164117 ; RRID:Addgene_164117) |
| 参考文献部分: | An expanded palette of improved SPLICS reporters detects multiple organelle contacts in vitro and in vivo. Vallese F, Catoni C, Cieri D, Barazzuol L, Ramirez O, Calore V, Bonora M, Giamogante F, Pinton P, Brini M, Cali T. Nat Commun. 2020 Nov 27;11(1):6069. doi: 10.1038/s41467-020-19892-6. 10.1038/s41467-020-19892-6 PubMed 33247103 |
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 SPLICS Mt-ER- Po Long P2A 7733 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Detect the long Peroxisomes-mitochondria-Endoplasmic reticulum
contact .
ACCESSION .
VERSION .
KEYWORDS SPLICS Mt-ER- Po Long P2A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7733)
AUTHORS Vallese F, Catoni C, Cieri D, Barazzuol L, Ramirez O, Calore V,
Bonora M, Giamogante F, Pinton P, Brini M, Cali T
TITLE An expanded palette of improved SPLICS reporters detects multiple
organelle contacts in vitro and in vivo.
JOURNAL Nat Commun. 2020 Nov 27;11(1):6069. doi: 10.1038/s41467-020-19892-6.
PUBMED 33247103
REFERENCE 2 (bases 1 to 7733)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7733
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(49..68)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 240..619
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 620..823
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 774..794
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 868..887
/label=T7
/note="T7 promoter, forward primer"
promoter 868..886
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 925..972
/codon_start=1
/product="11th beta-strand of superfolder GFP"
/label=GFP11
/note="associates with GFP(1-10), consisting of the first
10 beta-strands, to reconstitute fluorescent GFP (Cabantous
et al., 2005; Kamiyama et al., 2016)"
/translation="RDHMVLHEYVNAAGIT"
CDS 1489..1545
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 2335..2391
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
primer_bind complement(3329..3346)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3335..3559
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3605..4033
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3692..3711)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3902..3923
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4042..4062)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4047..4376
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4227..4362
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4289..4308
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4443..5237
/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(4497..4516)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5107..5126
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5411..5532
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5448..5467)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5502..5521
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5581..5597)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5581..5597)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5594..5616)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5605..5621
/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(5629..5659)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5674..5695
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5812..5829)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5983..6571)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6063..6082)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6742..7602)
/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 7365..7384
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7603..7707)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
901 ctagcgttta aacttaagct tatgcgggac cacatggtgc tgcacgagta cgtgaacgcc
961 gccggcatca ccggaggcga tggaggctcc ggaggaggct ctaagctgaa caattctggc
1021 gcccctcaca gggagaagag gggaggagag accgacgagt tctccaatgt gcggagaggc
1081 cggggccaca gaatgcagca cctgtctgag ggcacaaagg gaaggcaagt gggctctgga
1141 ggcgacggag agagatgggg cagcgatagg ggctctcgcg gcagcctgaa cgagcagatc
1201 gccctggtgc tgatgaggct gcaggaggat atgcagaatg tgctgcagcg cctgcagaag
1261 ctggagaccc tgacagccct gcaggccaag agctccacca gcacactgca gaccgcacca
1321 cagcctacaa gccagaggcc atcctggtgg ccctttgaga tgtcccctgg cgtgctgaca
1381 ttcgccatca tctggccctt catcgcccag tggctggtgt atctgtacta tcagaggcgc
1441 cggagaaagc tgaacggatc cctgaaggag ctggcatgcg gcagcggagc aaccaacttc
1501 tccctgctga agcaggcagg cgacgtggag gagaatcccg gccctcggtc tgaattcatg
1561 gtgggccgca attccgcaat cgcagcagga gtgtgcggcg ccctgttcat cggctactgt
1621 atctattttg accggaagcg gagaagcgat ccaaattcca agggcgagga gctgttcaca
1681 ggcgtggtgc ccatcctggt ggagctggac ggcgatgtga acggccacaa gttttctgtg
1741 cggggagagg gagagggcga cgcaaccatc ggcaagctga cactgaagtt catctgcacc
1801 acaggcaagc tgcccgtgcc ttggccaacc ctggtgacca cactgacata cggcgtgcag
1861 tgttttagca ggtatcctga ccacatgaag cgccacgatt tctttaagtc cgccatgcca
1921 gagggctacg tgcaggagcg gaccatctct ttcaaggacg atggcaagta taagaccaga
1981 gccgtggtga agtttgaggg cgacacactg gtgaacagga tcgagctgaa gggcaccgac
2041 ttcaaggagg atggcaatat cctgggccac aagctggagt acaactttaa tagccacaac
2101 gtgtatatca cagccgataa gcagaagaac ggcatcaagg ccaatttcac cgtgagacac
2161 aatgtggagg acggctccgt gcagctggcc gatcactacc agcagaacac accaatcggc
2221 gacggccctg tgctgctgcc tgataatcac tatctgtcta cccagacagt gctgagcaag
2281 gaccccaacg agaagggcac cctcgagctg aaggagctgg catgcggcag cggagcaacc
2341 aacttctccc tgctgaagca ggcaggcgac gtggaggaga atcccggccc tcggtctggg
2401 cccatgagca agggagagga gctgttcacc ggagtggtgc ctatcctggt ggagctggac
2461 ggcgatgtga atggccacaa gttttccgtg cgcggagagg gagagggcga cgcaacaatc
2521 ggcaagctga ccctgaagtt catctgcacc acaggcaagc tgcccgtgcc ttggccaacc
2581 ctggtgacca cactgggcta cggcctgcag tgttttgccc ggtatccaga ccacatgaag
2641 agacacgatt tctttaagtc cgccatgccc gagggctacg tgcaggagag gacaatctct
2701 ttcaaggacg atggcaagta taagacacgc gccgtggtga agtttgaggg cgacaccctg
2761 gtgaaccgga tcgagctgaa gggcacagac ttcaaggagg atggcaatat cctgggccac
2821 aagctggagt acaactttaa ttcccacaac gtgtatatca ccgccgataa gcagaagaac
2881 ggcatcaagg ccaatttcac agtgcggcac aatgtggagg acggctctgt gcagctggcc
2941 gatcactacc agcagaacac ccctatcggc gacggccctg tgctgctgcc tgataatcac
3001 tacctgtctt atcagaccgt gctgagcaag gacccaaacg agaagggcac aggaggcgat
3061 ggaggcagcg gaggaggctc caagctgaga gtgttcctgg ccctgcccat catcatggtg
3121 gtggcctttt ctatgtgcat catctgtctg ctgatggcag gcgacacctg gacagagacc
3181 ctggcctacg tgctgttttg gggcgtggcc agcatcggca cattctttat catcctgtat
3241 aacggcaagg acttcgtgga tgcccccagg ctggtgcaga aggagaagat cgattgatct
3301 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
3361 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
3421 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
3481 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
3541 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
3601 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
3661 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
3721 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
3781 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
3841 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
3901 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
3961 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
4021 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
4141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
4201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
4261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
4321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
4381 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
4441 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
4501 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
4561 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
4621 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
4681 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
4741 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
4801 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
4861 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
4921 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
4981 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
5041 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
5101 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
5161 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
5221 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
5281 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
5341 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
5401 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
5461 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
5521 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
5581 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
5641 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
5701 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
5761 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
5821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
5881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
5941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
6001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
6061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
6121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
6181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
6241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
6301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
6361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
6421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
6481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
6541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
6601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
6661 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
6721 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
6781 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
6841 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
6901 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
6961 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
7021 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
7081 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
7141 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
7201 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
7261 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
7321 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
7381 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
7441 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
7501 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
7561 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
7621 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
7681 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
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