| 质粒名称: | mAzurite-8xbiRhoBAST-24xMS2v6 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 196352 |
| 存储实验室: | Murat Sunbul |
|
载体骨架基本信息 |
|
| 载体名称: | mAzurite-C1 |
| 完整质粒大小: | 7790 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | mAzurite-8xbiRhoBAST-24xMS2 |
| 物种来源: | Synthetic |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知 (未破坏) |
|
文献引用方法 |
|
| 材料和方法部分: | mAzurite-8xbiRhoBAST-24xMS2v6 was a gift from Murat Sunbul (Addgene plasmid # 196352 ; http://n2t.net/addgene:196352 ; RRID:Addgene_196352) |
| 参考文献部分: | Bright, fluorogenic and photostable avidity probes for RNA imaging. Bühler B, Benderoth A, Englert D, Grün F, Schokolowski J, Jäschke A, Sunbul M. bioRxiv 2021 10.1101/2021.11.02.466936 |
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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 mAzurite-8xbiRhoBAST-24xMS2v6 7790 bp ds-DNA circular SYN 15-FEB-2023
DEFINITION Expression of mAzurite mRNA with 8xbiRhoBAST-24xMS2v6 in 3'UTR.
ACCESSION .
VERSION .
KEYWORDS mAzurite-8xbiRhoBAST-24xMS2v6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7790)
AUTHORS Buhler B, Benderoth A, Englert D, Grun F, Schokolowski J, Jaschke A,
Sunbul M
TITLE Bright, fluorogenic and photostable avidity probes for RNA imaging
JOURNAL bioRxiv 2021
REFERENCE 2 (bases 1 to 7790)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 15, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7790
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 133..436
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 437..640
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 591..611
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 679..688
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 685..1401
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNIYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFRTAAGITHGMDELYK"
primer_bind complement(730..751)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(991..1010)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
polyA_signal 4650..4771
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4687..4706)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4741..4760
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(4778..5233)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4915..4936)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5127..5146
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5260..5364
/gene="bla"
/label=AmpR promoter
promoter 5366..5723
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(5389..5409)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 5574..5709
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5636..5655
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5758..6552
/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(5812..5831)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6422..6441
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(6740..6759)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 6784..6831
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 7160..7748
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7649..7668
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
61 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
121 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
601 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
661 ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
841 aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt gcagtgcttc
901 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
1081 gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca caacatctat
1141 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
1201 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
1261 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
1321 aacgagaagc gcgatcacat ggtcctgctg gagttccgca ccgccgccgg gatcactcac
1381 ggcatggacg agctgtacaa gtccggactc agatctcgac gggtctagag gtatgtggga
1441 ggaacctccg cgaaagcggt gaaggagagg cgcaaggtta accgcctcag gttccaggag
1501 gattcgtcct ccaaaggcct ccgcgaaagc ggtgaaggag cgccacaagg ttaactggcg
1561 caggccttac ccacataccg ctagaggagc ccggaaggaa cctccgcgaa agcggtgaag
1621 gagaggcgca aggttaaccg cctcaggttc cagcggagtt cgctccgcaa aggcctccgc
1681 gaaagcggtg aaggagcgcc acaaggttaa ctggcgcagg ccttatccgg gctccgctag
1741 aggtatgtgg gaggaacctc cgcgaaagcg gtgaaggaga ggcgcaaggt taaccgcctc
1801 aggttccagg aggattcgtc ctccaaaggc ctccgcgaaa gcggtgaagg agcgccacaa
1861 ggttaactgg cgcaggcctt acccacatac cgctagagga gcccggaagg aacctccgcg
1921 aaagcggtga aggagaggcg caaggttaac cgcctcaggt tccagcggag ttcgctccgc
1981 aaaggcctcc gcgaaagcgg tgaaggagcg ccacaaggtt aactggcgca ggccttatcc
2041 gggctccgct agaggtatgt gggaggaacc tccgcgaaag cggtgaagga gaggcgcaag
2101 gttaaccgcc tcaggttcca ggaggattcg tcctccaaag gcctccgcga aagcggtgaa
2161 ggagcgccac aaggttaact ggcgcaggcc ttacccacat accgctagag gagcccggaa
2221 ggaacctccg cgaaagcggt gaaggagagg cgcaaggtta accgcctcag gttccagcgg
2281 agttcgctcc gcaaaggcct ccgcgaaagc ggtgaaggag cgccacaagg ttaactggcg
2341 caggccttat ccgggctccg ctagaggtat gtgggaggaa cctccgcgaa agcggtgaag
2401 gagaggcgca aggttaaccg cctcaggttc caggaggatt cgtcctccaa aggcctccgc
2461 gaaagcggtg aaggagcgcc acaaggttaa ctggcgcagg ccttacccac ataccgctag
2521 aggagcccgg aaggaacctc cgcgaaagcg gtgaaggaga ggcgcaaggt taaccgcctc
2581 aggttccagc ggagttcgct ccgcaaaggc ctccgcgaaa gcggtgaagg agcgccacaa
2641 ggttaactgg cgcaggcctt atccgggctc cgctagagcg gacttcggtc cgctttttac
2701 taggacctgc aggcatgcaa gcttgacgtc ggttaccgat atccatatgg cggccgcatc
2761 gatctcgagc tcaagcttcg aattctgcag tcgacgggcc cgggatccga tccaatcatg
2821 gatcccagag ccccctggca atcgcgggga gacgcaggac taccgcgtct tcactctcgc
2881 ttgcgcggta tacgcaggga gcaacgtcac ccagggcgac acagaggaat accctgtgtg
2941 gccccacggt gctacaaaga acattccgta ttgctcacca gtgtcacatg tggaggacta
3001 ccccacaagc gagtcaggaa accttcgggg catcgcacca ttatcccgaa caatcgacac
3061 gcaggattac cgcgtgggac actctgttcc ccgtcaaaat tggaccatac cggagtcggc
3121 ttacgtcatg caggattacc gcatgcatgg tgcagaatat cggcatgtct actcgtacag
3181 tcaaatctac tcgtgtggtc aggactaccg accacttcta ttctattcat cttttcggtt
3241 gtgcaggtat tctgccgatg tacgagaaga cgattacgct tctcgactac cttcatcata
3301 catggtgtgc agatggccgc caagtttttt gccaatggag gaatacccca ttctctgtca
3361 accaacccat gcaaagttta cactctgcta tggcagcact gtcgcagagg aataccctgc
3421 gagccaaaac ggcccccggt gcgtgtattg catctgcctt gcgagcattc acagggacga
3481 atacgccctg ccgttgcatt acttcaatat gggtgctctg tcgtcgtcat caggaccatt
3541 tgcgcaggac taccgcgcat atatcatcag cactcgtgcg gatacttctg ggattcctat
3601 tgttacgcga gctcaggaat accgagctct ggcgacagag accctcacac ggaagatccc
3661 agagccccct ggcaatcgcg gggagacgca ggactaccgc gtcttcactc tcgcttgcgc
3721 ggtatacgca gggagcaacg tcacccaggg cgacacagag gaataccctg tgtggcccca
3781 cggtgctaca aagaacattc cgtattgctc accagtgtca catgtggagg actaccccac
3841 aagcgagtca ggaaaccttc ggggcatcgc accattatcc cgaacaatcg acacgcagga
3901 ttaccgcgtg ggacactctg ttccccgtca aaattggacc ataccggagt cggcttacgt
3961 catgcaggat taccgcatgc atggtgcaga atatcggcat gtctactcgt acagtcaaat
4021 ctactcgtgt ggtcaggact accgaccact tctattctat tcatcttttc ggttgtgcag
4081 gtattctgcc gatgtacgag aagacgatta cgcttctcga ctaccttcat catacatggt
4141 gtgcagatgg ccgccaagtt ttttgccaat ggaggaatac cccattctct gtcaaccaac
4201 ccatgcaaag tttacactct gctatggcag cactgtcgca gaggaatacc ctgcgagcca
4261 aaacggcccc cggtgcgtgt attgcatctg ccttgcgagc attcacaggg acgaatacgc
4321 cctgccgttg cattacttca atatgggtgc tctgtcgtcg tcatcaggac catttgcgca
4381 ggactaccgc gcatatatca tcagcactcg tgcggatact tctgggattc ctattgttac
4441 gcgagctcag gaataccgag ctctggcgac agagaccctc acacggaaga tctatcgatc
4501 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
4561 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
4621 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
4681 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
4741 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
4801 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
4861 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
4921 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
4981 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
5041 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
5101 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
5161 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
5221 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
5281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
5341 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
5401 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
5461 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
5521 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
5581 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
5641 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
5701 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
5761 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
5821 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
5881 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
5941 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
6001 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
6061 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
6121 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
6181 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
6241 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
6301 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
6361 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
6421 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
6481 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
6541 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
6601 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
6661 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
6721 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
6781 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
6841 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
6901 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
6961 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
7021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
7081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
7141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
7201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
7261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
7321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
7381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
7441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
7501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
7561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
7621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
7681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
7741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
//
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