LOCUS       Exported                6394 bp ds-DNA    circular SYN 12-8-2013
DEFINITION  Dual-promoter mammalian vector for strong constitutive expression of
            a gene together with firefly luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    pSF-CMV-PGK-FLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6394)
  AUTHORS   Oxford Genetics
  TITLE     Direct Submission
COMMENT     The PGK promoter is typically about 10-fold weaker than the CMV 
            promoter.
FEATURES             Location/Qualifiers
     source          1..6394
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     misc_feature    18..89
                     /note="5 beta-globin insulator"
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     enhancer        287..590
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        591..794
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    857..942
                     /note="MCS"
                     /note="multiple cloning site"
     RBS             893..898
                     /note="Shinbe-Dalgarno ribosome binding site"
     misc_signal     903..909
                     /note="Kozak sequence"
     CDS             907..909
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     misc_feature    961..971
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     promoter        988..1487
                     /note="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1498..3150
                     /codon_start=1
                     /product="firefly luciferase"
                     /note="luciferase"
                     /note="synthetic luc2 version of the luciferase gene"
                     /protein_id="
                     "
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    3158..3168
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    3200..3321
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     terminator      3436..3482
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3512..3648
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3693..3764
                     /note="3 beta-globin insulator"
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3990..4578)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     RBS             5311..5316
                     /note="Shine-Dalgarno ribosome binding site"
     CDS             5324..6118
                     /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"
     terminator      6131..6267
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"