LOCUS       Exported                5475 bp ds-DNA    circular SYN 12-8-2013
DEFINITION  Bacterial vector for expressing firefly luciferase from a strong 
            constitutive recA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pSF-RecA-FLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5475)
  AUTHORS   Oxford Genetics
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5475
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /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)"
     promoter        359..427
                     /note="recA (Delta-LexA) promoter"
                     /note="constitutively active E. coli recA promoter lacking 
                     the LexA binding site (Brent and Ptashne, 1981)"
     RBS             505..510
                     /note="Shine-Dalgarno ribosome binding site"
     misc_signal     515..521
                     /note="Kozak sequence"
     CDS             519..2171
                     /codon_start=1
                     /product="firefly luciferase"
                     /note="luciferase"
                     /note="synthetic luciferase gene"
                     /protein_id="
                     "
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    2191..2201
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     misc_feature    2239..2249
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    2281..2402
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     terminator      2517..2563
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2593..2729
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2774..2845
                     /note="3 beta-globin insulator"
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3071..3659)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     RBS             4392..4397
                     /note="Shine-Dalgarno ribosome binding site"
     CDS             4405..5199
                     /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      5212..5348
                     /note="rrnG terminator"
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"