| 质粒名称: | pRSET-B mCherry |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 108857 |
| 存储实验室: | Kalina Hristova |
|
载体骨架基本信息 |
|
| 载体名称: | pRSET-B |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mCherry |
| 物种来源: | Discosoma (coral) |
| 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: | pRSET-B mCherry was a gift from Kalina Hristova (Addgene plasmid # 108857 ; http://n2t.net/addgene:108857 ; RRID:Addgene_108857) |
| 参考文献部分: | Uninduced high-yield bacterial expression of fluorescent proteins. Sarabipour S, King C, Hristova K. Anal Biochem. 2014 Mar 15;449:155-7. doi: 10.1016/j.ab.2013.12.027. Epub 2013 Dec 28. 10.1016/j.ab.2013.12.027 PubMed 24378720 |
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 pRSET-B mCherry 3577 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Encodes for the fluorescent protein mCherry.
ACCESSION .
VERSION .
KEYWORDS pRSET-B mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3577)
AUTHORS Sarabipour S, King C, Hristova K
TITLE Uninduced high-yield bacterial expression of fluorescent proteins.
JOURNAL Anal Biochem. 2014 Mar 15;449:155-7. doi: 10.1016/j.ab.2013.12.027.
Epub 2013 Dec 28.
PUBMED 24378720
REFERENCE 2 (bases 1 to 3577)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3577
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 9..597
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 498..517
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 751..768
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 853..872
/label=T7
/note="T7 promoter, forward primer"
promoter 853..871
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 903..925
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 945..962
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 966..998
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 1002..1025
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
regulatory 1032..1041
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1038..1748
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(1183..1201)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1451..1470
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1811..1829)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1825..1872
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1969..2424
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2056..2075)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2266..2287
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 2450..2554
/gene="bla"
/label=AmpR promoter
CDS 2555..3415
/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 complement(2773..2792)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
781 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggatctcg
841 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
901 attttgttta actttaagaa ggagatatac atatgcgggg ttctcatcat catcatcatc
961 atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg
1021 ataaggatcc cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
1081 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
1141 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
1201 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
1261 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
1321 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
1381 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
1441 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
1501 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
1561 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
1621 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
1681 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
1741 acaagtaaga attcgaagct tgatccggct gctaacaaag cccgaaagga agctgagttg
1801 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
1861 aggggttttt tgctgaaagg aggaactata tccggatctg gcgtaatagc gaagaggccc
1921 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
1981 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
2041 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
2101 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
2161 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
2221 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
2281 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
2341 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
2401 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
2461 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
2521 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
2581 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
2641 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
2701 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
2761 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
2821 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
2881 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
2941 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
3001 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
3061 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
3121 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
3181 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
3241 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
3301 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
3361 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
3421 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
3481 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
3541 gttccactga gcgtcagacc ccgtagaaaa gatcaaa
//
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