LOCUS       Exported                6064 bp ds-DNA    circular SYN 26-1-2014
DEFINITION  IRES-containing bicistronic vector for expressing a gene together 
            with the AcGFP1 fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    pEF1alpha-IRES-AcGFP1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  AUTHORS   Clontech
  TITLE     Direct Submission
COMMENT     The gene inserted into the MCS should contain start and stop codons.
FEATURES             Location/Qualifiers
     source          1..6064
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     promoter        165..1346
                     /note="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          395..1337
                     /note="EF-1-alpha intron A"
                     /note="intron upstream of the start codon"
     misc_feature    1366..1422
                     /note="MCS"
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     misc_feature    1424..2010
                     /note="IRES2"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1999..2001
                     /codon_start=1
                     /product="start codon for translation from IRES2"
                     /note="ATG"
                     /translation="M"
     CDS             2011..2730
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /note="AcGFP1"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2852..2973
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2980..3435)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3462..3566
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        3568..3925
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      3776..3911
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3960..4754
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4986..5033
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      5362..5950
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"