pIRES2- DsRed- Express Plasmid
PVT1229 2ug
pIRES2- DsRed- Express Plasmid Informaiton
Promoter: CMV
Replicon: pUC ori, F1 ori
Terminator: SV40 poly (A) signal
Plasmid classification: mammalian cells, fluorescent protein reporter vectors
Plasmid size: 5264bp
Prokaryotic resistance: Kan
Selection marker: Neo
Clonal strain: DH5 alpha
Culture conditions: 37 C, aerobic LB
Expression host: lactation cells
Induction mode: no need to induce, transient expression.
5'sequencing primers: CMV-F:CGCAAATGGGCGGTAGGCGTG
Primers for 3'sequencing: primers designed based on sequences
pIRES2- DsRed- Express Plasmid Description
pIRES2- DsRed- Express Plasmid contains the internal ribosome entry site (IRES; 1, 2) of the encephalomyocarditis virus (ECMV) between the MCS and a mutant of the Discosoma sp. red fluorescent protein DsRed-Express coding region (3–5). This permits both the gene of interest (cloned into the MCS) and the DsRed-Express gene to be translated from a single bicistronic mRNA. pIRES2-DsRed-Express is designed for the efficient selection by flow cytometry of transiently transfected mammalian cells expressing DsRed-Express and the protein of interest. This vector can also be used to express DsRed-Express alone or to obtain stably transfected cell lines without timeconsuming drug and clonal selection.
pIRES2- DsRed- Express Plasmid Multiple cloning site
pIRES2- DsRed- Express Plasmid Sequence
LOCUS Exported 5264 bp ds-DNA circular SYN 01-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5264)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, September 1, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..5264
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 609..665
/note="MCS"
/note="multiple cloning site"
misc_feature 667..1253
/note="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 1254..1931
/codon_start=1
/product="rapidly maturing tetrameric variant of DsRed
fluorescent protein (Bevis and Glick, 2002)"
/note="DsRed-Express"
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
LFL"
polyA_signal 2052..2173
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2180..2635)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2662..2766
/gene="bla"
/note="AmpR promoter"
promoter 2768..3125
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2976..3111
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 3160..3954
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 4186..4233
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4562..5150
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
661 gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
721 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
781 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
841 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
901 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
961 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
1021 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
1081 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
1141 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
1201 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggcct
1261 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga
1321 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
1381 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
1441 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca
1501 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
1561 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg ctccttcatc tacaaggtga
1621 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct
1681 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca
1741 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtct atctatatgg
1801 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct
1861 cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc
1921 tgttcctgta gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
1981 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
2041 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
2101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
2161 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
2221 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
2281 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
2341 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
2401 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
2461 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
2521 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
2581 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
2641 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
2701 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
2761 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
2941 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
3001 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
3061 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
3121 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
3181 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
3241 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
3301 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
3361 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
3421 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
3481 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
3541 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
3601 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
3661 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
3721 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
3781 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
3841 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
3901 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
3961 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
4021 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
4081 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
4141 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
4201 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
4261 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
4321 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
4381 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
4441 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
4501 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
4561 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
4621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
4681 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
4741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
4801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
4861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
4921 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
4981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
5041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
5101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
5161 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
5221 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
Caution:
1. This product is FOR RESEARCH USE ONLY!
2. The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.