Induction mode: no need to induce, transient expression.
pIRES2-EGFP contains the internal ribosome entry site (IRES; 1, 2) of the encephalomyocarditis virus (ECMV) between the MCS and the enhanced green fluorescent protein (EGFP) coding region.This permits both the gene of interest (cloned into the MCS) and the EGFP gene to be translated from a single bicistronic mRNA. pIRES2-EGFP is designed for the efficient selection (by flow cytometry or other methods) of transiently transfected mammalian cells expressing EGFP and the protein of interest. This vector can also be used to express EGFP alone or to obtain stably transfected cell lines without time-consuming drug and clonal selection.
LOCUS Exported 5308 bp ds-DNA circular SYN 31-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pIRES2-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5308)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-10-22
REFERENCE 2 (bases 1 to 5308)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5308
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/note="CMV enhancer"
/note="CMV enhancer;human cytomegalovirus immediate early
enhancer"
promoter 365..568
/note="CMV promoter"
/note="CMV promoter;human cytomegalovirus (CMV) immediate
early promoter"
misc_feature 609..665
/note="MCS"
/note="MCS;multiple cloning site"
misc_feature 667..1253
/note="IRES2"
/note="IRES2;internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 1254..1973
/codon_start=1
/product="enhanced GFP"
/note="enhanced GFP"
/note="EGFP;mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 2096..2217
/note="SV40 poly(A) signal"
/note="SV40 poly(A) signal;SV40 polyadenylation signal"
rep_origin complement(2224..2679)
/direction=LEFT
/note="f1 ori"
/note="f1 ori;f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2706..2810
/gene="bla"
/note="bla AmpR promoter"
/note="AmpR promoter"
promoter 2812..3169
/note="SV40 promoter"
/note="SV40 promoter;SV40 enhancer and early promoter"
rep_origin 3020..3155
/note="SV40 ori"
/note="SV40 ori;SV40 origin of replication"
CDS 3204..3998
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="aph(3')-II (or nptII)"
/note="NeoR/KanR;confers resistance to neomycin, kanamycin,
and G418 (Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 4230..4277
/note="HSV TK poly(A) signal"
/note="HSV TK poly(A) signal;herpesvirus thymidine kinase
polyadenylation signal"
rep_origin 4606..5194
/direction=RIGHT
/note="ori"
/note="ori;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 accatggtga
1261 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
1321 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
1381 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
1441 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
1501 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
1561 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
1621 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
1681 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
1741 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
1801 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
1861 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
1921 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
1981 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
2041 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
2101 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
2161 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
2221 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
2281 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
2341 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
2401 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
2461 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
2521 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
2581 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
2641 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
2701 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
2761 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
2821 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
2881 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
3001 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
3061 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
3121 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
3181 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
3241 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
3301 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
3361 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
3421 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
3481 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
3541 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
3601 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
3661 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
3721 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
3781 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
3841 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
3901 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
3961 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
4021 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
4081 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
4141 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
4201 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
4261 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
4321 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
4381 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
4441 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
4501 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
4561 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
4621 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
4681 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
4741 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
4801 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
4861 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
4921 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
4981 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
5041 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
5101 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
5161 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
5221 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
5281 tgtggataac cgtattaccg ccatgcat
//
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