LOCUS       Exported                4945 bp ds-DNA    circular SYN 12-8-2013
DEFINITION  Vector encoding a minimal promoter and Renilla luciferase, for 
            measuring the activity of transcriptional regulatory elements.
ACCESSION   .
VERSION     .
KEYWORDS    pSF-pA-MinProm-RLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4945)
  AUTHORS   Oxford Genetics
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4945
                     /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)"
     polyA_signal    339..387
                     /note="synthetic polyadenylation signal"
     misc_feature    401..492
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     misc_feature    493..530
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        535..597
                     /note="Mini-TK promoter"
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     RBS             689..694
                     /note="Shine-Dalgarno ribosome binding site"
     misc_signal     699..705
                     /note="Kozak sequence"
     CDS             703..1641
                     /codon_start=1
                     /product="Renilla luciferase"
                     /note="RLuc"
                     /note="codon-optimized synthetic gene"
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQG"
     misc_feature    1661..1671
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     misc_feature    1709..1719
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    1751..1872
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     terminator      1987..2033
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2063..2199
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2244..2315
                     /note="3 beta-globin insulator"
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2541..3129)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     RBS             3862..3867
                     /note="Shine-Dalgarno ribosome binding site"
     CDS             3875..4669
                     /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      4682..4818
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"