| 质粒名称: | Str-KDEL_ManII-SBP-mCherry |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 65253 |
| 存储实验室: | Franck Perez |
|
载体骨架基本信息 |
|
| 载体名称: | pIRES_neo3 |
| 载体出品公司: | Clontech |
| 空载体大小: | -- |
| 完整质粒大小: | 6272 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 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: | 链亲和素KDEL与截短型甘露糖苷酶II |
| 物种来源: | M.musculus（小鼠） |
| 插入大小: | -- |
| 基因突变: | 仅含有ManII的氨基酸1-116 |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | 链亲和素结合蛋白（SBP）（插入物上的C末端）mCherry（插入件上的C端子） |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | IRES-F |
| 3’测序引物 | mCherry-R |
|
文献引用方法 |
|
| 材料和方法部分: | Str-KDEL_ManII-SBP-mCherry was a gift from Franck Perez (Addgene plasmid # 65253 ; http://n2t.net/addgene:65253 ; RRID:Addgene_65253) |
| 参考文献部分: | Synchronization of secretory protein traffic in populations of cells. Boncompain G, Divoux S, Gareil N, de Forges H, Lescure A, Latreche L, Mercanti V, Jollivet F, Raposo G, Perez F. Nat Methods. 2012 Mar 11;9(5):493-8. doi: 10.1038/nmeth.1928. 10.1038/nmeth.1928 PubMed 22406856 |
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 Str-KDEL_ManII-SBP-mCherry 6272 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synchronize trafficking of ManII from the ER (RUSH system).
ACCESSION .
VERSION .
KEYWORDS Str-KDEL_ManII-SBP-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6272)
AUTHORS Boncompain G, Divoux S, Gareil N, de Forges H, Lescure A, Latreche
L, Mercanti V, Jollivet F, Raposo G, Perez F
TITLE Synchronization of secretory protein traffic in populations of
cells.
JOURNAL Nat Methods. 2012 Mar 11;9(5):493-8. doi: 10.1038/nmeth.1928.
PUBMED 22406856
REFERENCE 2 (bases 1 to 6272)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6272
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
intron 1646..1875
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
primer_bind 1858..1877
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature 1933..2506
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2100..2117)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2327..2346
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2908..3021
/codon_start=1
/product="streptavidin-binding peptide"
/label=SBP
/note="selected from a peptide library; binds streptavidin
with nanomolar affinity (Keefe et al., 2001)"
/translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
CDS 3031..3738
/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(3176..3194)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3444..3463
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 3892..4013
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3929..3948)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4115..4131)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4115..4131)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4128..4150)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4139..4155
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4163..4193)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4208..4229
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4346..4363)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4517..5105)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4597..4616)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5276..6136)
/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 5899..5918
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6137..6241)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgacc
1081 ctagcaaaga ctcaaaagct caggtgtccg ctgccgaggc tggcattact ggaacatggt
1141 acaatcagct cgggagcacc tttattgtga ctgctggagc cgatggagcc ctcaccggaa
1201 catacgaatc tgctgtggga aacgccgaat cacggtacgt cctcactggc cgatacgata
1261 gtgcccctgc caccgacgga tctgggactg ccctgggatg gactgtcgct tggaaaaaca
1321 actaccggaa tgctcattct gccacaacat ggagtggaca gtacgtggga ggcgctgagg
1381 ctagaatcaa tacacagtgg ctgctcacat ctggcacaac cgaggcaaat gcttggaaat
1441 ccaccctggt gggacatgac acattcacca aagtgaaacc ctccgccgct tcaatcgatg
1501 ccgccaaaaa agccggagtc aacaacggca atcctctgga tgccgtccag cagctcggac
1561 cgaaagatga actgtgagcg gccgcataga taactgatcc agtgtgctgg aattaattcg
1621 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
1681 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
1741 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
1801 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
1861 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
1921 cggccaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
1981 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
2041 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
2101 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
2161 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
2221 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
2281 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
2341 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
2401 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
2461 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
2521 gggaggcgcg ccatgaagtt aagtcgccag ttcaccgtgt ttggcagcgc gatcttctgc
2581 gtcgtaatct tctcactcta cctgatgctg gacaggggtc acttggacta ccctcggggc
2641 ccgcgccagg agggctcctt tccgcagggc cagctttcaa tattgcaaga aaagattgac
2701 catttggagc gtttgctcgc tgagaacaac gagattatct caaatatcag agactcagtc
2761 atcaacctga gcgagtctgt ggaggacggc ccgcgggggt caccaggcaa cgccagccaa
2821 ggctccatcc acctccactc gccacagttg gccctgcagg ctgaccccag agactgtttg
2881 gatcccaccg gtcgccaccg gaattccatg gacgagaaga ccactggttg gcgaggtgga
2941 cacgttgttg aaggactggc tggggaactt gaacaacttc gtgcacgact ggagcatcac
3001 ccacaaggtc aacgtgaacc acctgcaggt atggtgagca agggcgagga ggataacatg
3061 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
3121 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
3181 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
3241 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
3301 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
3361 accgtgaccc aggactcctc cctacaggac ggcgagttca tctacaaggt gaagctgcgc
3421 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
3481 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
3541 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
3601 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
3661 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
3721 atggacgagc tgtacaaggg ccggccttaa ggcctcgagg gccaattaat taactctaga
3781 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
3841 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
3901 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
3961 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgtc
4021 gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
4081 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
4141 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
4201 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
4261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
4321 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
4381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
4441 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
4501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
4561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
4621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
4681 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
4741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
4801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
4861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
4921 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
4981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
5041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
5101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
5161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
5221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
5281 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
5341 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
5401 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
5461 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
5521 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
5581 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
5641 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
5701 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
5761 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
5821 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
5881 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
5941 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
6001 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
6061 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
6121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
6181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
6241 gcacatttcc ccgaaaagtg ccacctgacg tc
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：WT PBRM1- TRIPZ-neo
- 下一篇：pJRH029
