LOCUS       Exported                6510 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pFUSE-HEAVY, complete sequence.
ACCESSION   KC176268
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6510)
  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  JOURNAL   Unpublished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 6510)
  AUTHORS   Loppnau PA, Graslund S, Arrowsmith CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-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 6510)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6510
                     /organism="Expression vector pFUSE-HEAVY"
                     /mol_type="other DNA"
                     /db_xref="taxon:1294006"
     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    573..632
                     /label=human interleukin-2 secretion signal
                     /note="human interleukin-2 secretion signal"
     CDS             573..632
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     misc_feature    628..642
                     /label=5-In-Fusion cloning site
                     /note="5-In-Fusion cloning site"
     misc_feature    633..642
                     /note="partial coding sequence human immunoglobulin heavy 
                     chain variable region"
     protein_bind    657..690
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        705..1150
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1151..2572
                     /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="AGH39947.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            1151..2572
                     /gene="sacB"
                     /label=sacB
     misc_feature    2634..2652
                     /note="partial coding sequence human immunoglobulin heavy 
                     chain variable region"
     misc_feature    2634..2648
                     /label=3-In-Fusion cloning site
                     /note="3-In-Fusion cloning site"
     misc_feature    2653..2949
                     /note="human immunoglobulin gamma 1 heavy chain constant 1 
                     region"
     misc_feature    2950..2994
                     /label=human immunoglobulin gamma 1 hinge region
                     /note="human immunoglobulin gamma 1 hinge region"
     misc_feature    2995..3324
                     /note="mouse immunoglobulin gamma 1 heavy chain constant 2 
                     region"
     misc_feature    3325..3645
                     /note="mouse immunoglobulin gamma 1 heavy chain constant 3 
                     region"
     regulatory      3669..3886
                     /regulatory_class="polyA_signal_sequence"
                     /note="simian virus 40 late polyadenylation signal"
     polyA_signal    3675..3796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3900..4300)
                     /regulatory_class="polyA_signal_sequence"
                     /note="human beta globin polyadenylation signal"
     polyA_site      3900..4294
                     /label=-beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4308..4682)
                     /codon_start=1
                     /gene="ble"
                     /product="phleomycin/zeocin binding protein"
                     /label=ble
                     /note="phleomycin/zeocin resistance"
                     /protein_id="AGH39946.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     gene            complement(4308..4682)
                     /gene="ble"
                     /label=ble
                     /note="Sh ble"
     regulatory      complement(4732..4787)
                     /regulatory_class="promoter"
                     /note="EM2KC bacterial promoter"
     regulatory      complement(4905..5167)
                     /regulatory_class="promoter"
                     /note="human ferritin light chain mammalian cell promoter"
     regulatory      complement(5174..5578)
                     /regulatory_class="enhancer"
                     /note="human cytomegalovirus immediate-early gene 1 
                     enhancer"
     enhancer        5275..5578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5658..6246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(5659)
                     /direction=LEFT
                     /label=colE1 from pBR322
                     /note="colE1 from pBR322"
     polyA_signal    6354..6402
                     /note="synthetic polyadenylation signal"
     misc_feature    6413..6504
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"