| 质粒名称: | PSD-95-pTagRFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 52671 |
| 存储实验室: | Johannes Hell |
|
载体骨架基本信息 |
|
| 载体名称: | pTagRFP |
| 载体出品公司: | Evrogen |
| 空载体大小: | 4733 bp |
| 完整质粒大小: | 6904 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin |
| 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: | PSD-95 / Postsynaptic density protein 95 / Disks large homolog 4 (DLG4) / Synapse-associated protein 90 (SAP-90) |
| 物种来源: | R. norvegicus (rat) |
| GenBank编号 | P78352 |
| 启动子: | CMV |
| 标签/融合蛋白: | tagRFP (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HindIII (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | mKate2-TurboRFP-R |
|
文献引用方法 |
|
| 材料和方法部分: | PSD-95-pTagRFP was a gift from Johannes Hell (Addgene plasmid # 52671 ; http://n2t.net/addgene:52671 ; RRID:Addgene_52671) |
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 PSD-95-pTagRFP 6904 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION expression of TagRFP-tagged PSD-95.
ACCESSION .
VERSION .
KEYWORDS PSD-95-pTagRFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6904)
TITLE Hell lab plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6904)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6904
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 54..357
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 358..561
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 512..532
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 621..630
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 2844..2853
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2850..3563
/codon_start=1
/product="monomeric derivative of red fluorescent protein
from Entacmaea quadricolor (Merzlyak et al., 2007)"
/label=TagRFP
/note="mammalian codon-optimized"
/translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
ARYCDLPSKLGHKLN"
polyA_signal 3685..3806
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3722..3741)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3776..3795
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3813..4268)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3950..3971)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4162..4181
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4295..4399
/gene="bla"
/label=AmpR promoter
promoter 4401..4758
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4424..4444)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4609..4744
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4671..4690
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4793..5587
/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(4847..4866)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5457..5476
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5775..5794)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5819..5866
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6195..6783
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6684..6703
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
601 cagatctcga gctcaagctt gccaccatgg actgtctctg tatagtgaca accaagaaat
661 accgctacca agatgaagac acgccccctc tggaacacag cccggcccac ctccccaacc
721 aggccaattc tccccctgtg attgtcaaca cggacaccct agaagcccca ggatatgagt
781 tgcaggtgaa tggaacagag ggggagatgg agtatgagga gatcacattg gaaaggggta
841 actcaggtct gggcttcagc atcgcaggtg gcactgacaa cccgcacatc ggtgacgacc
901 cgtccatttt tatcaccaag atcattcctg gtggggctgc agcccaggat ggccgcctca
961 gggtcaatga cagcatcctg tttgtaaatg aagtggatgt tcgggaggtg acccattcag
1021 ctgcggtgga ggccctcaaa gaggcaggtt ccatcgttcg cctctatgtc atgcgccgga
1081 aacccccagc cgaaaaggtc atggagatca aactcatcaa agggcctaaa ggacttggct
1141 tcagcattgc ggggggcgtt gggaaccagc acatccctgg agataacagc atctatgtaa
1201 cgaagatcat cgaaggaggt gctgcccaca aggatggcag gttgcagatt ggagacaaga
1261 tcctggcggt caacagtgtg gggctggagg acgtcatgca cgaggatgcc gtggcagccc
1321 tgaagaacac atatgacgtt gtgtacctaa aggtggccaa gcccagcaat gcctacctga
1381 gtgacagcta tgctccccca gacatcacaa cctcgtattc tcagcacctg gacaatgaga
1441 tcagtcatag cagctacttg ggcactgact accccacagc catgaccccc acttcccctc
1501 ggcgctactc ccctgtggcc aaggacctgc tgggggagga agacattccc cgggaaccaa
1561 ggcggatcgt gatccatcgg ggctccaccg gcctgggctt caacatcgtg ggcggcgagg
1621 atggtgaagg catcttcatc tccttcatcc ttgctggggg tccagccgac ctcagtgggg
1681 agctacggaa gggggaccag atcctgtcgg tcaatggtgt tgacctccgc aatgccagtc
1741 acgaacaggc tgccattgcc ctgaagaatg cgggtcagac ggtcacgatc atcgctcagt
1801 ataaaccaga agagtatagt cgattcgagg ccaagatcca tgatcttcgg gaacagctca
1861 tgaatagtag cctaggctca gggactgcat ccttgcgaag caaccccaag aggggcttct
1921 acattagggc cctgtttgat tacgacaaga ccaaggactg cggtttcttg agccaggccc
1981 tgagcttccg cttcggggat gtgcttcatg tcattgacgc tggtgacgaa gagtggtggc
2041 aagcacggcg ggtccactcc gacagtgaga ccgacgacat tggcttcatt cccagcaaac
2101 ggcgggtcga gcgacgagag tggtcaaggt taaaggccaa ggactggggc tccagctctg
2161 gatcacaggg tcgagaagac tcggttctga gctatgagac ggtgacccag atggaagtgc
2221 actatgctcg tcccatcatc atccttggac ccaccaaaga ccgtgccaac gatgatcttc
2281 tctccgagtt ccccgacaag tttggatcct gtgtccctca tacgacacgt cctaagcggg
2341 aatatgagat agacggccgg gattaccact ttgtctcctc ccgggagaaa atggagaagg
2401 acatccaggc acacaagttc attgaggctg gccagtacaa cagccacctc tatgggacca
2461 gcgtccagtc tgtgcgagag gtagcagagc aggggaagca ctgcatcctc gatgtctcgg
2521 ccaatgccgt gcggcggctg caggcggccc acctgcaccc catcgccatc ttcatccgtc
2581 cccgctccct ggagaatgtg ctagagatca ataagcggat cacagaggag caagcccgga
2641 aagccttcga cagagccacg aagctggagc aggagttcac agagtgcttc tcagccatcg
2701 tagagggcga cagctttgaa gagatctatc acaaagtgaa acgtgtcatt gaagacctct
2761 caggccccta catctgggtc ccagcccgag agagactcgg aattctgcag tcgacggtac
2821 cgcgggcccg ggatccaccg gtcgccacca tggtgtctaa gggcgaagag ctgattaagg
2881 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
2941 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
3001 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
3061 gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg
3121 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
3181 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
3241 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
3301 tgtaccccgc tgacggcggc ctggaaggca gaagcgacat ggccctgaag ctcgtgggcg
3361 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
3421 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
3481 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
3541 aactggggca caaacttaat tgagcggccg cgactctaga tcataatcag ccataccaca
3601 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
3661 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
3721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
3781 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
3841 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
3901 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
3961 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
4021 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
4081 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
4141 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
4201 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
4261 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
4321 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
4381 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
4441 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
4501 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
4561 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
4621 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
4681 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
4741 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
4801 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
4861 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
4921 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
4981 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
5041 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
5101 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
5161 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
5221 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
5281 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
5341 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
5401 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
5461 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
5521 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
5581 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
5641 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
5701 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
5761 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
5821 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
5881 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
5941 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
6001 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
6061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
6841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
6901 ttat
//
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