Plasmid classification: lactation serial plasmids; lactating fluorescent plasmid; lactation green plasmid
pEGFP-C2 Plasmid encodes a red-shifted variant of wild-type GFP which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) pEGFP-C2 encodes the GFPmut1 variant which contains the double-amino-acid substitution of Phe-64 to Leu and Ser-65 to Thr. The coding sequence of the EGFP gene contains more than 190 silent base changes which correspond to human codon-usage preferences . Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site to further increase the translation efficiency in eukaryotic cells. The MCS in pEGFP-C2 is between the EGFP coding sequences and the SV40 poly A. Genes cloned into the MCS will be expressed as fusions to the C terminus of EGFP if they are in the same reading frame as EGFP and there are no intervening stop codons. SV40 polyadenylation signals downstream of the EGFP gene direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen. A neomycin-resistance cassette (Neor), consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene, allows stably transfected eukaryotic cells to be selected using G418. A bacterial promoter upstream of this cassette expresses kanamycin resistance in E. coli. The pEGFP-C2 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production. Fusions to the C terminus of EGFP retain the fluorescent properties of the native protein allowing the localization of the fusion protein in vivo. The target gene should be cloned into pEGFP-C2 so that it is in frame with the EGFP coding sequences, with no intervening in-frame stop codons. The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418. pEGFP-C2 can also be used simply to express EGFP in a cell line of interest (e.g., as a transfection marker). Suitable host strains: DH5α, HB101, and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM109 or XL1-Blue. Selectable marker: plasmid confers resistance to kanamycin (30 µg/ml) to E. coli hosts.
LOCUS Exported 4735 bp ds-DNA circular SYN 31-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pEGFP-C2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4735)
TITLE Direct Submission
JOURNAL Exported 2015-5-31
FEATURES Location/Qualifiers
source 1..4735
/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"
CDS 613..1329
/codon_start=1
/product="enhanced GFP"
/note="enhanced GFP"
/note="EGFP;mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 1343..1399
/note="MCS"
/note="MCS;multiple cloning site"
polyA_signal 1523..1644
/note="SV40 poly(A) signal"
/note="SV40 poly(A) signal;SV40 polyadenylation signal"
rep_origin complement(1651..2106)
/direction=LEFT
/note="f1 ori"
/note="f1 ori;f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2133..2237
/gene="bla"
/note="bla AmpR promoter"
/note="AmpR promoter"
promoter 2239..2596
/note="SV40 promoter"
/note="SV40 promoter;SV40 enhancer and early promoter"
rep_origin 2447..2582
/note="SV40 ori"
/note="SV40 ori;SV40 origin of replication"
CDS 2631..3425
/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 3657..3704
/note="HSV TK poly(A) signal"
/note="HSV TK poly(A) signal;herpesvirus thymidine kinase
polyadenylation signal"
rep_origin 4033..4621
/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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1321 ctgtacaagt ccggccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg
1381 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
1441 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
1501 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
1561 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
1621 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
1681 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
1741 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
1801 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
1861 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
1921 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
1981 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
2041 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
2101 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
2161 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
2221 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
2281 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
2341 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
2401 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
2461 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
2521 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
2581 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
2641 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
2701 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
2761 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
2821 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
2881 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
2941 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
3001 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
3061 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
3121 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
3181 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
3241 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
3301 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
3361 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
3421 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
3481 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
3541 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
3601 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
3661 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
3721 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
3781 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
3841 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
3901 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
3961 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
4021 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
4081 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
4141 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
4201 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
4261 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
4321 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
4381 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
4441 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
4501 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
4561 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
4621 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
4681 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
Publication for PVT1206 pEGFP-C2
Article: Ubiquitin specific peptidase 1 promotes hepatic fibrosis through positive regulation of CXCL1 by deubiquitinating SNAIL
Author: ZhiyongDu a TianchongWu a LinsenLiu a BiweiLuo a CuifengWei b
Digestive and Liver Disease Available online 27 April 2021
Received 19 November 2020, Accepted 26 February 2021, Available online 27 April 2021.
DOI: https://doi.org/10.1016/j.dld.2021.02.025
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.
Search name
pEGFP-C2,Plasmid pEGFP-C2,pEGFP-C2 vector
English