pEGFP- Actin Plasmid

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  • Model: PVT1212
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pEGFP-Actin

PVT1212    2ug
 

pEGFP-Actin Information

romoter: CMV

Replicator: pUC ori, F1 ori

Terminator: SV40 poly (A) signal

Plasmid classification: mammalian cells, fluorescent protein reporting carriers

Plasmid size: 5820bp

Prokaryotic resistance: Kan

Screening markers: Neo

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic LB

Expression host: mammalian cells

Induction mode: no induction, instantaneous expression

5'sequencing primers: CMV-F:CGCAAATGGGCGG

 

pEGFP-Actin Description

pEGFP-Actin encodes a fusion protein consisting of the red-shifted, human codon-optimized variant of green fluorescent protein (EGFP; 1–3) and the gene encoding human cytoplasmic b-actin (4). EGFP, a derivative of the GFPmut1 variant (5), has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) This variant contains the double-amino-acid substitution of Phe-64 to Leu and Ser-65 to Thr, and also contains more than 190 silent base changes which correspond to human codon-usage preferences (6). SV40 polyadenylation signals downstream of the EGFP-Actin fusion direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in any mammalian cell line that expresses 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 drives expression of the gene encoding kanamycin resistance in E. coli. The pEGFPActin backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production

pEGFP-Actin-polyclonal-site

 

pEGFP-Actin Sequence

LOCUS       Exported                5820 bp ds-DNA    circular SYN 18-10-2015

DEFINITION  .

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 3

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 5820)

  AUTHORS   admin

  TITLE     Direct Submission

  JOURNAL   Exported 2015-10-18  

FEATURES             Location/Qualifiers

     source          1..5820

                     /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"

     CDS             613..1329

                     /codon_start=1

                     /product="enhanced GFP"

                     /note="EGFP"

                     /note="mammalian codon-optimized"

                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL

                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD

                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK

                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL

                     EFVTAAGITLGMDELYK"

     polyA_signal    2608..2729

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(2736..3191)

                     /direction=LEFT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        3218..3322

                     /gene="bla"

                     /note="AmpR promoter"

     promoter        3324..3681

                     /note="SV40 promoter"

                     /note="SV40 enhancer and early promoter"

     rep_origin      3532..3667

                     /note="SV40 ori"

                     /note="SV40 origin of replication"

     CDS             3716..4510

                     /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    4742..4789

                     /note="HSV TK poly(A) signal"

                     /note="herpesvirus thymidine kinase polyadenylation signal"

     rep_origin      5118..5706

                     /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 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 ccggactcag atctcgagct atggatgatg atatcgccgc gctcgtcgtc

     1381 gacaacggct ccggcatgtg caaggccggc ttcgcgggcg acgatgcccc ccgggccgtc

     1441 ttcccctcca tcgtggggcg ccccaggcac cagggcgtga tggtgggcat gggtcagaag

     1501 gattcctatg tgggcgacga ggcccagagc aagagaggca tcctcaccct gaagtacccc

     1561 atcgagcacg gcatcgtcac caactgggac gacatggaga aaatctggca ccacaccttc

     1621 tacaatgagc tgcgtgtggc tcccgaggag caccccgtgc tgctgaccga ggcccccctg

     1681 aaccccaagg ccaaccgcga gaagatgacc cagatcatgt ttgagacctt caacacccca

     1741 gccatgtacg ttgctatcca ggctgtgcta tccctgtacg cctctggccg taccactggc

     1801 atcgtgatgg actccggtga cggggtcacc cacactgtgc ccatctacga ggggtatgcc

     1861 ctcccccatg ccatcctgcg tctggacctg gctggccggg acctgactga ctacctcatg

     1921 aagatcctca ccgagcgcgg ctacagcttc accaccacgg ccgagcggga aatcgtgcgt

     1981 gacattaagg agaagctgtg ctacgtcgcc ctggacttcg agcaagagat ggccacggct

     2041 gcttccagct cctccctgga gaagagctac gagctgcctg acggccaggt catcaccatt

     2101 ggcaatgagc ggttccgctg ccctgaggca ctcttccagc cttccttcct gggcatggag

     2161 tcctgtggca tccacgaaac taccttcaac tccatcatga agtgtgacgt ggacatccgc

     2221 aaagacctgt acgccaacac agtgctgtct ggcggcacca ccatgtaccc tggcattgcc

     2281 gacaggatgc agaaggagat cactgccctg gcacccagca caatgaagat caagatcatt

     2341 gctcctcctg agcgcaagta ctccgtgtgg atcggcggct ccatcctggc ctcgctgtcc

     2401 accttccagc agatgtggat cagcaagcag gagtatgacg agtccggccc ctccatcgtc

     2461 caccgcaaat gcttctaggg atccaccgga tctagataac tgatcataat cagccatacc

     2521 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa

     2581 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa

     2641 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt

     2701 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt

     2761 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg

     2821 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg

     2881 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta

     2941 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg

     3001 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa

     3061 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct

     3121 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct

     3181 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt

     3241 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca

     3301 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag

     3361 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc

     3421 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa

     3481 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc

     3541 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat

     3601 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt

     3661 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat

     3721 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta

     3781 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca

     3841 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga

     3901 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga

     3961 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct

     4021 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg

     4081 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga

     4141 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca

     4201 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga

     4261 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg

     4321 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc

     4381 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt

     4441 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga

     4501 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca

     4561 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc

     4621 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac

     4681 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg

     4741 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg

     4801 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac

     4861 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc

     4921 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag

     4981 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat

     5041 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa

     5101 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca

     5161 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt

     5221 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg

     5281 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc

     5341 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga

     5401 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc

     5461 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc

     5521 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca

     5581 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg

     5641 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta

     5701 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct

     5761 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat

//

 

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-Actin,Plasmid pEGFP-Actin,pEGFP-Actin vector

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