LOCUS       Exported                5680 bp ds-DNA    circular SYN 12-8-2013
DEFINITION  Dual-promoter mammalian vector for strong constitutive expression of
            a gene together with Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    pSF-CMV-PGK-RLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  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..5680
                     /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="Shine-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..2436
                     /codon_start=1
                     /product="Renilla luciferase"
                     /note="RLuc"
                     /note="codon-optimized synthetic gene"
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQG"
     misc_feature    2444..2454
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    2486..2607
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     terminator      2722..2768
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2798..2934
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2979..3050
                     /note="3 beta-globin insulator"
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3276..3864)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     RBS             4597..4602
                     /note="Shine-Dalgarno ribosome binding site"
     CDS             4610..5404
                     /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      5417..5553
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"