pFUSE-HEAVY vector (V006153)

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LOCUS       40924_20721        6510 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pFUSE-HEAVY, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6510)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-NOV-2012) Structural Genomics Consortium, University of Toronto,
            Mars Centre, South Tower, 101 College St, Room 700, Toronto, Ontario
            M5G 1L7, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     sig_peptide     573..632
                     /label=IL-2 signal sequence
                     /note="human interleukin-2 signal sequence"
     misc_feature    633..642
                     /note="partial coding sequence human immunoglobulin heavy
                     chain variable region"
     protein_bind    complement(657..690)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        705..1150
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1151..2569
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     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"
     polyA_signal    complement(3675..3796)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    complement(3900..4294)
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(4311..4682)
                     /label=BleoR
                     /note="antibiotic-binding protein"
     regulatory      complement(4732..4787)
                     /label=EM2KC bacterial promoter
                     /note="EM2KC bacterial promoter"
                     /regulatory_class="promoter"
     regulatory      complement(4905..5167)
                     /label=human ferritin light chain mammalian cell promoter
                     /note="human ferritin light chain mammalian cell promoter"
                     /regulatory_class="promoter"
     enhancer        complement(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"
     polyA_signal    6354..6402
                     /label=poly(A) signal
                     /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"