LOCUS       Exported                5983 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pFUSE-LIGHT, complete sequence.
ACCESSION   KC176267
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5983)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     A pipeline to generate recombinant antibodies to human target 
            proteins involved in epigenetic mechanisms
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5983)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-NOV-2012) Structural Genomics Consortium, University 
            of Toronto, Mars Centre, South Tower, 101 College St, Room 700, 
            Toronto, Ontario M5G 1L7, Canada
REFERENCE   3  (bases 1 to 5983)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5983
                     /organism="Expression vector pFUSE-LIGHT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1294005"
     regulatory      12..256
                     /regulatory_class="promoter"
                     /note="elongation factor-1 alpha mammalian cell promoter"
     promoter        31..242
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             255..523
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     5UTR           257..544
                     /note="human T-cell leukemia virus type 1 long terminal 
                     repeat"
     misc_feature    556..615
                     /label=human interleukin-2 secretion signal
                     /note="human interleukin-2 secretion signal"
     CDS             556..615
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     misc_feature    616..753
                     /note="partial coding sequence human Immunoglobulin light 
                     chain, kappa type"
     misc_feature    694..708
                     /label=5-In-Fusion cloning site
                     /note="5-In-Fusion cloning site"
     protein_bind    765..798
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        813..1258
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1259..2680
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /note="Bacillus subtilis negative selection gene; converts 
                     sucrose to a toxic product; allows selection of recombinant
                     plasmids on media containing 5% sucrose"
                     /protein_id="AGH39945.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            1259..2680
                     /gene="sacB"
                     /label=sacB
     misc_feature    2737..2757
                     /note="partial coding sequence human Immunoglobulin light 
                     chain, kappa type"
     misc_feature    2742..2757
                     /label=3-In-Fusion cloning site
                     /note="3-In-Fusion cloning site"
     misc_feature    2758..3081
                     /note="coding sequence of human Immunoglobulin light 
                     constant chain"
     regulatory      3100..3333
                     /regulatory_class="polyA_signal_sequence"
                     /note="simian virus 40 late polyadenylation signal"
     polyA_signal    3118..3239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3342..3744)
                     /regulatory_class="polyA_signal_sequence"
                     /note="human beta globin polyadenylation signal"
     polyA_site      3343..3737
                     /label=-beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(3746..4168)
                     /codon_start=1
                     /gene="bsrS2"
                     /product="blasticidin S-deaminase"
                     /label=bsrS2
                     /note="Bacillus cereus blasticidin resistance gene"
                     /protein_id="AGH39944.1"
                     /translation="MKTFNISQQDLELVEVATEKITMLYEDNKHHVGAAIRTKTGEIIS
                     AVHIEAYIGRVTVCAEAIAIGSAVSNGQKDFDTIVAVRHPYSDEVDRSIRVVSPCGMCR
                     ELISDYAPDCFVLIEMNGKLVKTTIEELIPLKYTRN"
     gene            complement(3746..4168)
                     /gene="bsrS2"
                     /label=bsrS2
     regulatory      complement(4218..4260)
                     /regulatory_class="promoter"
                     /note="EM2KC bacterial promoter"
     regulatory      complement(4378..4640)
                     /regulatory_class="promoter"
                     /note="human ferritin light chain mammalian cell promoter"
     regulatory      complement(4647..5051)
                     /regulatory_class="enhancer"
                     /note="human cytomegalovirus immediate-early gene 1 
                     enhancer"
     enhancer        4748..5051
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5131..5719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(5132)
                     /direction=LEFT
                     /label=colE1 from pBR322
                     /note="colE1 from pBR322"
     polyA_signal    5827..5875
                     /note="synthetic polyadenylation signal"
     misc_feature    5886..5977
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"