| 质粒名称: | pmTurquoise2-Golgi |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 36205 |
| 存储实验室: | Dorus Gadella |
|
载体骨架基本信息 |
|
| 载体名称: | pmTurquoise2-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4904bp |
| 完整质粒大小: | 4904bp |
| 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: | Golgi targeting sequence / B4GALT1(1-61) /beta4Gal-T1(1-61)/ beta-1,4-galactosyltransferase 1(1-61) |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 183bp |
| GenBank编号 | NM_001497 |
| 启动子: | CMV |
| 标签/融合蛋白: | mTurquoise2 (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII (not destroyed) |
| 3’克隆位点: | AgeI (not destroyed) |
| 5’测序引物 | AGGTCTATATAAGCAGAGC |
| 3’测序引物 | TCTACAAATGTGGTATGGC |
|
文献引用方法 |
|
| 材料和方法部分: | pmTurquoise2-Golgi was a gift from Dorus Gadella (Addgene plasmid # 36205 ; http://n2t.net/addgene:36205 ; RRID:Addgene_36205) |
| 参考文献部分: | Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen L, Hink MA, van Weeren L, Gadella TW Jr, Royant A. Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738. 10.1038/ncomms1738 PubMed 22434194 |
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 pmTurquoise2-Golgi 4904 bp ds-DNA circular SYN 17-JUN-2022
DEFINITION In vivo visualization of golgi apparatus (can be used for
colocalization studies).
ACCESSION .
VERSION .
KEYWORDS pmTurquoise2-Golgi
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4904)
AUTHORS Goedhart J, von Stetten D, Noirclerc-Savoye M, Lelimousin M, Joosen
L, Hink MA, van Weeren L, Gadella TW Jr, Royant A
TITLE Structure-guided evolution of cyan fluorescent proteins towards a
quantum yield of 93%.
JOURNAL Nat Commun. 2012 Mar 20;3:751. doi: 10.1038/ncomms1738.
PUBMED 22434194
REFERENCE 2 (bases 1 to 4904)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 17, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4904
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 9..312
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 313..516
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 467..487
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 792..801
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 798..1517
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2012)"
/label=mTurquoise2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(843..864)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1104..1123)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1451..1472
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1640..1761
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1677..1696)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1731..1750
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1768..2223)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1905..1926)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2117..2136
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2250..2354
/gene="bla"
/label=AmpR promoter
promoter 2356..2713
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2379..2399)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2564..2699
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2626..2645
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2748..3542
/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(2802..2821)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3412..3431
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3730..3749)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3774..3821
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4150..4738
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4639..4658
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
481 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
541 tagcgctacc ggactcagat ctcgagctcg ccaccatgag gcttcgggag ccgctcctga
601 gcggcagcgc cgcgatgcca ggcgcgtccc tacagcgggc ctgccgcctg ctcgtggccg
661 tctgcgctct gcaccttggc gtcaccctcg tttactacct ggctggccgc gacctgagcc
721 gcctgcccca actggtcgga gtctccacac cgctgcagtc gacggtaccg cgggcccggg
781 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
841 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
901 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
961 ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct
1021 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
1081 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
1141 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
1201 gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc tatatcaccg
1261 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
1321 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
1381 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
1441 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
1501 acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
1561 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
1621 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
1681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
1741 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
1801 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
1861 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
1921 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
1981 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
2041 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
2101 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
2161 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
2221 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
2281 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
2341 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
2401 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
2461 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
2521 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
2581 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
2641 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
2701 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
2761 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
2821 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
2881 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
2941 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
3001 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
3061 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
3121 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
3181 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
3241 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
3301 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
3361 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
3421 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
3481 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
3541 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
3601 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
3661 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
3721 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
3781 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
3841 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
3901 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
3961 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
4021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
4081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
4141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
4201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
4261 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
4321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
4381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
4441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
4501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
4561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
4621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
4681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
4741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
4801 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
4861 aatagtaatc aattacgggg tcattagttc atagcccata tatg
//
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