pFB-LIC-Bse vector (V010743)

LOCUS       pFB-LIC-Bse.        6816 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Baculovirus vector encoding an N-terminal 6xHis-TEV cassette and 
            ampicillin resistance plus a SacB negative selection marker, for 
            purification of recombinant proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pFB-LIC-Bse
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  AUTHORS   Savitsky P, Bray J, Cooper CD, Marsden BD, Mahajan P, Burgess-Brown 
            NA, Gileadi O.
  TITLE     High-throughput production of human proteins for crystallization: 
            the SGC experience.
  JOURNAL   J. Struct. Biol. 2010;172:3-13.
  PUBMED    20541610
REFERENCE   2  (bases 1 to 6816)
  AUTHORS   Structural Genomics Consortium
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6816)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Struct. 
            Biol."; date: "2010"; volume: "172"; pages: "3-13"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     For ligation-independent cloning (LIC), linearize with BseRI to 
            remove the SacB cassette, and treat with T4 DNA polymerase plus 
            dGTP.
FEATURES             Location/Qualifiers
     source          1..6816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             148..150
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             154..171
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             196..216
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     protein_bind    complement(246..279)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        294..739
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             740..2158
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     polyA_signal    2403..2537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  complement(2566..2731)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2915..3370
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3397..3501
                     /label=AmpR promoter
     CDS             3502..4359
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4533..5121
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  complement(5424..5648)
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5718..6248)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(6437..6465)
                     /label=Pc promoter
                     /note="class 1 integron promoter"