| 质粒名称: | pEF.myc.ER-E2-Crimson |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 38770 |
| 存储实验室: | Benjamin Glick |
|
载体骨架基本信息 |
|
| 载体名称: | pEF/myc/ER |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 5603 bp |
| 完整质粒大小: | 6263 bp |
| 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: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | E2-Crimson,Modified DsRed-Express2 |
| 物种来源: | Synthetic |
| 插入大小: | 675 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | EF-1a |
| 标签/融合蛋白: |
ER信号序列（MGWSCIILFLVATATGAHS）（主干上的N端）
myc（主干上的C终端）
ER保持信号（KDEL）（主干上的C终端）
|
| 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 | EF1a_fwd |
| 3’测序引物 | BGH_rev |
|
文献引用方法 |
|
| 材料和方法部分: | pEF.myc.ER-E2-Crimson was a gift from Benjamin Glick (Addgene plasmid # 38770 ; http://n2t.net/addgene:38770 ; RRID:Addgene_38770) |
| 参考文献部分: | A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling. Strack RL, Hein B, Bhattacharyya D, Hell SW, Keenan RJ, Glick BS. Biochemistry. 2009 Sep 8;48(35):8279-81. 10.1021/bi900870u PubMed 19658435 |
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 pEF.myc.ER-E2-Crimson 6269 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION In vivo visualization of the endoplasmic reticulum (can be used for
colocalization studies).
ACCESSION .
VERSION .
KEYWORDS pEF.myc.ER-E2-Crimson
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6269)
AUTHORS Strack RL, Hein B, Bhattacharyya D, Hell SW, Keenan RJ, Glick BS
TITLE A rapidly maturing far-red derivative of DsRed-Express2 for
whole-cell labeling.
JOURNAL Biochemistry. 2009 Sep 8;48(35):8279-81.
PUBMED 19658435
REFERENCE 2 (bases 1 to 6269)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6269
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 96..231
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 158..177
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 271..1065
/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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(325..344)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 935..954
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 1239..1360
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1276..1295)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1330..1349
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1393..1410)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1564..2152)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1644..1663)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2323..3183)
/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 2946..2965
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3184..3288)
/gene="bla"
/label=AmpR promoter
promoter 3364..4526
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 3595..4517
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4474..4494
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 4534..4543
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
sig_peptide join(4540..4584,4667..4678)
/product="modified signal sequence from the mouse
immunoglobulin heavy chain variable region"
/label=IgH signal sequence
CDS 4700..5374
/codon_start=1
/product="far-red noncytotoxic tetrameric variant of DsRed
fluorescent protein (Strack et al., 2009)"
/label=E2-Crimson
/note="mammalian codon-optimized"
/translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
LFQ"
primer_bind 5098..5117
/label=mCherry-F
/note="mCherry, forward primer"
CDS 5384..5413
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(5467..5484)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5473..5697
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5743..6171
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5830..5849)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6040..6061
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6180..6200)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6185..245
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
ORIGIN
1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
121 ccgcccattc tccgccccta ggctgactaa ttttttttat ttatgcagag gccgaggccg
181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
241 gcaaaaagct ccccccccgg gaggtccaca atggttgaac aagatggatt gcacgcaggt
301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
421 accgacctgt ccggtgccct gaatgaactc caggacgagg cagcgcggct atcgtggctg
481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
781 ttcgccaggc tcaaggcgcg tatgcccgac ggcgaggatc tcgtcgtgac tcatggcgat
841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
1021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
1081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
1141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
1201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
1261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
1321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
1381 tctttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
1801 ctccaagctg ggctgtatgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
1981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
3061 tcgggcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
3301 aaaagtgcca cctgacgtca gatcgacgga tcgggagatc ggtaccgaat tcaagcttcg
3361 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
3421 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
3481 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
3541 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
3601 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
3661 attacttcca cctggctcca gtacgtgatt cttgatcccg agctggagcc aggggcgggc
3721 cttgcgcttt aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
3781 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
3841 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
3901 taaatgcggg ccaggatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
3961 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
4021 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
4081 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
4141 aaagatggcc gcttcccggc cctgctccag ggggctcaaa atggaggacg cggcgctcgg
4201 gagagcgggc gggtgagtca cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg
4261 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctggagct
4321 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
4381 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
4441 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
4501 tttttcttcc atttcaggtg tcgtgaacac gtggccacca tgggatggag ctgtatcatc
4561 ctcttcttgg tagcaacagc tacaggtaag gggttaacag tagcaggctt gaggtctgga
4621 catatatatg ggtgacaatg acatccactt tgcctttctc tccacaggcg cgcactccca
4681 ggtccaactg caggtcgaca tggatagcac tgagaacgtc atcaagccct tcatgcgctt
4741 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgtgggcga
4801 gggcaagccc tacgagggca cccagaccgc caagctgcaa gtgaccaagg gcggccccct
4861 gcccttcgcc tgggacatcc tgtcccccca gttcttctac ggctccaagg cgtacatcaa
4921 gcaccccgcc gacatccccg actacctcaa gcagtccttc cccgagggct tcaagtggga
4981 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
5041 ggacggcacc ctcatctacc acgtgaagtt catcggcgtg aacttcccct ccgacggccc
5101 cgtaatgcag aagaagactc tgggctggga gccctccact gagcgcaact acccccgcga
5161 cggcgtgctg aagggcgaga accacatggc gctgaagctg aagggcggcg gccactacct
5221 gtgtgagttc aagtccatct acatggccaa gaagcccgtg aagctgcccg gctaccacta
5281 cgtggactac aagctcgaca tcacctccca caacgaggac tacaccgtgg tggagcagta
5341 cgagcgcgcc gaggcccgcc accacctgtt ccaggcggcc gcagaacaaa aactcatctc
5401 agaagaggat ctgaatgggg ccgcaagcga gaaggacgag ctgtagtcta gaagctcgct
5461 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
5521 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
5581 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
5641 agggggagga ttgggaagac aatagcaggc atgctgggga tggcccgggc tctatggctt
5701 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
5761 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
5821 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
5881 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctct
5941 cccccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
6001 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
6061 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
6121 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
6181 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
6241 aagcatgcat ctcaattagt cagcaacca
//
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