| 质粒名称: | mCherry-ER-3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 55041 |
| 存储实验室: | Michael Davidson |
|
载体骨架基本信息 |
|
| 载体名称: | mCherry |
| 载体出品公司: | -- |
| 空载体大小: | 4750 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 卡那霉素 |
| 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: | ER,Calreticulin + KDEL,NM_004343.3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | ER Signal Peptide |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | mCherry（主干上的C终端） |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | mCherry-ER-3 was a gift from Michael Davidson (Addgene plasmid # 55041 ; http://n2t.net/addgene:55041 ; RRID:Addgene_55041) |
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 mCherry-ER-3 4752 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Localization: Endoplasmic Reticulum, Excitation: 587, Emission: 610.
ACCESSION .
VERSION .
KEYWORDS mCherry-ER-3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4752)
TITLE Davidson lab - mCherry constructs
REFERENCE 2 (bases 1 to 4752)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4752
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 293..1000
/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(438..456)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 706..725
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 1176..1297
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1213..1232)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1267..1286
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1304..1759)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1441..1462)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1653..1672
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 1786..1890
/gene="bla"
/label=AmpR promoter
promoter 1892..2249
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(1915..1935)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2100..2235
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2162..2181
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2284..3078
/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(2338..2357)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2948..2967
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3266..3285)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3310..3357
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 3686..4274
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4175..4194
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 4449..4752
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatgctgct
241 atccgtgccg ttgctgctcg gcctcctcgg cctggccgtc gccgaccggt cgatggtgag
301 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
361 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
421 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
481 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
541 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
601 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
661 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
721 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
781 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
841 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
901 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
961 gggccgccac tccaccggcg gcatggacga gctgtacaag aaggacgagc tgtaagaatt
1021 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
1081 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
1141 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
1201 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
1261 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
1321 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
1381 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
1441 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
1501 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
1561 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
1621 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
1681 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
1741 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
1801 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
1861 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
1921 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
1981 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
2041 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
2101 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
2161 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
2221 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
2281 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
2341 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
2401 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
2461 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
2521 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
2581 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
2641 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
2701 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
2761 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
2821 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
2881 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
2941 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
3001 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
3061 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
3121 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
3181 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
3241 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
3301 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
3361 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
3421 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
3481 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
3541 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
3601 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
3661 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
3721 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
3781 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
3841 tgtagccgta gttaggccac cacttcaaga actctgtatc accgcctaca tacctcgctc
3901 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
3961 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
4021 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
4081 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
4141 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
4201 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
4261 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
4321 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
4381 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
4441 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
4501 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
4561 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
4621 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
4681 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
4741 gctattacca tg
//
