| 质粒名称: | hOCT1-miRFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 188995 |
| 存储实验室: | Jeremy Baskin |
|
载体骨架基本信息 |
|
| 载体名称: | miRFP-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4966 bp |
| 完整质粒大小: | 6628 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | hOCT1、SLC22A1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1662 bp |
| GenBank编号 | JN247409 |
| 启动子: | CMV |
| 标签/融合蛋白: | miRFP (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物 | CMV-S |
| 3’测序引物 | miRFP-AS |
|
文献引用方法 |
|
| 材料和方法部分: | |
| 参考文献部分: | |
买家导航
hOCT1-miRFP载体质粒基本信息
hOCT1-miRFP质粒图谱载体图谱和 hOCT1-miRFP载体序列质粒序列多克隆位点信息
hOCT1-miRFP质粒载体简介
hOCT1-miRFP质粒序列载体序列
LOCUS hOCT1-miRFP 6628 bp DNA circular SYN 14-APR-2025
DEFINITION A mammalian expression plasmid encoding miRFP-tagged human OCT1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6628)
AUTHORS Chiu DC, Baskin JM
TITLE Organelle-Selective Membrane Labeling through Phospholipase
D-Mediated Transphosphatidylation.
JOURNAL JACS Au. 2022 Nov 28;2(12):2703-2713. doi: 10.1021/jacsau.2c00419.
eCollection 2022 Dec 26.
PUBMED 36590261
REFERENCE 2 (bases 1 to 6628)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1021/jacsau.2c00419"; journalName: "JACS Au"; date: "2022-11-28-
28"; volume: "2"; issue: "12"; pages: "2703-2713"
COMMENT Sequence Label: hOCT1-miRFP
FEATURES Location/Qualifiers
source 1..6628
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 106..409
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 410..613
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 564..584
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 2386..2395
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2392..3339
/codon_start=1
/product="bright monomeric near-infrared fluorescent
protein engineered from a bacterial phytochrome, with an
emission maximum of 670 nm (Shcherbakova et al., 2016)"
/label=miRFP670
/label=ORF frame 1
/translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
VCHHYLPRFIRFELRAICKRLAERIATRITALES"
polyA_signal 3461..3582
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3498..3517)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3552..3571
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3589..4044)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3726..3747)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3938..3957
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4071..4175
/gene="bla"
/label=AmpR promoter
promoter 4177..4534
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 4177..4368
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(4200..4220)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4385..4520
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4569..5363
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 4569..5363
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4623..4642)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5233..5252
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5551..5570)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5595..5642
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5971..6559
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6460..6479
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
61 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
421 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
481 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
541 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
601 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
661 gaggccacca tgcccaccgt ggacgatatc ctggagcaag tgggcgagag cggctggttc
721 cagaagcagg cctttctgat cctgtgcctg ctgtccgccg ccttcgcccc tatctgcgtg
781 ggcatcgtgt tcctgggctt taccccagac caccactgcc agagcccagg agtggcagag
841 ctgtctcaga gatgtggctg gagcccagcc gaggagctga actacacagt gccaggactg
901 ggacctgcag gagaggcctt tctgggccag tgccggagat atgaggtgga ctggaaccag
961 tctgccctga gctgcgtgga tcctctggcc tccctggcca ccaataggtc tcacctgcca
1021 ctgggaccat gccaggacgg atgggtgtac gataccccag gcagctccat cgtgacagag
1081 ttcaacctgg tgtgcgccga ctcttggaag ctggatctgt ttcagagctg tctgaatgcc
1141 ggcttcctgt ttggcagcct gggcgtgggc tatttcgccg atagatttgg caggaagctg
1201 tgcctgctgg gaaccgtgct ggtgaacgcc gtgtccggcg tgctgatggc cttcagcccc
1261 aattacatgt ccatgctgct gtttcgcctg ctgcagggcc tggtgtctaa gggcaattgg
1321 atggccggct ataccctgat cacagagttc gtgggatccg gatctaggag gacagtggcc
1381 atcatgtacc agatggcctt taccgtggga ctggtggccc tgacaggact ggcatatgca
1441 ctgccacact ggaggtggct gcagctggcc gtgtccctgc ccaccttcct gtttctgctg
1501 tactattggt gcgtgcctga gagcccaaga tggctgctgt cccagaagag gaacacagag
1561 gccatcaaga tcatggacca catcgcccag aagaatggca agctgccccc tgccgacctg
1621 aagatgctga gcctggagga ggatgtgacc gagaagctga gcccctcctt cgccgacctg
1681 tttcggaccc ctcggctgag aaagaggaca ttcatcctga tgtacctgtg gtttaccgat
1741 tccgtgctgt atcagggcct gatcctgcac atgggcgcca catccggcaa cctgtacctg
1801 gacttcctgt attctgccct ggtggagatc ccaggcgcct ttatcgccct gatcacaatc
1861 gatcgcgtgg gccggatcta ccccatggca atgtctaacc tgctggcagg agcagcatgt
1921 ctggtcatga tcttcatcag ccccgacctg cactggctga atatcatcat catgtgcgtg
1981 ggccgcatgg gcatcaccat cgccatccag atgatctgtc tggtgaacgc cgagctgtat
2041 cctacatttg tgcggaatct gggcgtgatg gtgtgctcta gcctgtgcga tatcggcggc
2101 atcatcaccc ctttcatcgt gtttcgcctg agggaagtgt ggcaggccct gccactgatc
2161 ctgttcgccg tgctgggact gctggcagca ggagtgaccc tgctgctgcc tgagacaaag
2221 ggcgtggccc tgccagagac catgaaggat gccgagaacc tgggcagaaa ggccaagccc
2281 aaggagaata caatctacct gaaggtgcag acctccgagc cttctggcac aggaggctcc
2341 ggaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac catggtagca
2401 ggtcatgcct ctggcagccc cgcattcggg accgcctctc attcgaattg cgaacatgaa
2461 gagatccacc tcgccggctc gatccagccg catggcgcgc ttctggtcgt cagcgaacat
2521 gatcatcgcg tcatccaggc cagcgccaac gccgcggaat ttctgaatct cggaagcgta
2581 ctcggcgttc cgctcgccga gatcgacggc gatctgttga tcaagatcct gccgcatctc
2641 gatcccaccg ccgaaggcat gccggtcgcg gtgcgctgcc ggatcggcaa tccctctacg
2701 gagtactgcg gtctgatgca tcggcctccg gaaggcgggc tgatcatcga actcgaacgt
2761 gccggcccgt cgatcgatct gtcaggcacg ctggcgccgg cgctggagcg gatccgcacg
2821 gcgggttcac tgcgcgcgct gtgcgatgac accgtgctgc tgtttcagca gtgcaccggc
2881 tacgaccggg tgatggtgta tcgtttcgat gagcaaggcc acggcctggt attctccgag
2941 tgccatgtgc ctgggctcga atcctatttc ggcaaccgct atccgtcgtc gactgtcccg
3001 cagatggcgc ggcagctgta cgtgcggcag cgcgtccgcg tgctggtcga cgtcacctat
3061 cagccggtgc cgctggagcc gcggctgtcg ccgctgaccg ggcgcgatct cgacatgtcg
3121 ggctgcttcc tgcgctcgat gtcgccgtgc catctgcagt tcctgaagga catgggcgtg
3181 cgcgccaccc tggcggtgtc gctggtggtc ggcggcaagc tgtggggcct ggttgtctgt
3241 caccattatc tgccgcgctt catccgtttc gagctgcggg cgatctgcaa acggctcgcc
3301 gaaaggatcg cgacgcggat caccgcgctt gagagctaag cggccgcgac tctagatcat
3361 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
3421 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
3481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
3541 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
3601 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
3661 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
3721 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
3781 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
3841 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
3901 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
3961 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
4021 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
4081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
4141 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
4201 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
4261 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
4321 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
4381 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
4441 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
4501 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
4561 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
4621 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
4681 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
4741 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
4801 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
4861 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
4921 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
4981 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
5041 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
5101 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
5161 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
5221 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
5281 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
5341 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
5401 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
5461 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
5521 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
5581 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
5641 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
5701 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
5761 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
5821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
5881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
5941 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
6001 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
6061 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
6121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
6181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
6241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
6301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
6361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
6421 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
6481 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
6541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
6601 ctggcctttt gctcacatgt tctttcct
//
