pEYFP- Actin Plasmid

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  • Model: PVT1224
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pEYFP-Actin

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pEYFP-Actin,Plasmid pEYFP-Actin,pEYFP-Actin vector

 

pEYFP-Actin information

Promoter: 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:CGCAAATGGGCGGTAGGCGTG
3'sequencing primers: primers designed according to sequence

 

pEYFP-Actin Description

pEYFP-Actin encodes a fusion of the enhanced yellow fluorescent protein (EYFP) and human cytoplasmic β-actin. EYFP is an enhanced yellow-green variant of the Aequorea victoria green fluorescent protein (GFP). The EYFP gene contains the four amino acid substitutions previously published as GFP-10C: Ser-65 to Gly; Val-68 to Leu; Ser-72 to Ala; and Thr-203 to Tyr. It also contains the Phe-64 to Leu mutation of GFPmut1. The fluorescence excitation maximum of EYFP is 513 nm; the emission spectrum has a peak at 527 nm (in the yellow-green region). When excited at 513 nm, the Em of EYFP is 36,500 cm–1M–1 and the fluorescence quantum yield is 0.63, resulting in a bright fluorescent signal. The fluorescence observed is roughly equivalent to that from EGFP

pEYFP-Actin-polyclonal-site

 

pEYFP-Actin Sequence

LOCUS       Exported                5820 bp ds-DNA    circular SYN 19-10-2015
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported 2015-10-19   from MLCC
         
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 YFP"
                     /note="EYFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     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 caccttcggc tacggcctgc agtgcttcgc 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 ctaccagtcc 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 ggcgtaaatt gtaagcgtta atattttgtt
     2761 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2821 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
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