LOCUS       Exported                4575 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSCCA5-E8d DNA, complete sequence.
ACCESSION   AB969670
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4575)
  AUTHORS   Arimitsu H, Sasaki K, Iba Y, Kurosawa Y, Shimizu T, Tsuji T.
  TITLE     Isolation of B subunit-specific monoclonal antibody clones that 
            strongly neutralize the toxicity of Shiga toxin 2
  JOURNAL   Microbiol. Immunol. 59 (2), 71-81 (2015)
  PUBMED    25521016
REFERENCE   2  (bases 1 to 4575)
  AUTHORS   Arimitsu H, Iba Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2014) Contact:Hideyuki Arimitsu Fujita Health 
            University, Department of Microbiology; 1-98, Dengakugakubo, 
            Kutsukake-cho, Toyoake, Aichi 470-1192, Japan URL 
            :https://fujita-hu.ac.jp/
REFERENCE   3  (bases 1 to 4575)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4575
                     /organism="Expression vector pSCCA5-E8d"
                     /mol_type="other DNA"
                     /db_xref="taxon:1510416"
     misc_feature    1..1769
                     /label=single-chain Fv cloning region
                     /note="single-chain Fv cloning region"
     misc_feature    40..87
                     /label=PelB leader sequence
                     /note="PelB leader sequence"
     misc_feature    197..241
                     /label=Single chain linker
                     /note="Single chain linker"
     misc_feature    711..1340
                     /label=cp3
                     /note="cp3"
     misc_feature    1375..1755
                     /label=IgG binding domain of protein A
                     /note="IgG binding domain of protein A"
     CDS             1396..1566
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             1570..1740
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     misc_feature    1770..4575
                     /label=derived from pTZ19R DNA
                     /note="derived from pTZ19R DNA"
     primer_bind     complement(1770..1786)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1928..2383)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2410..2514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2515..3375
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3546..4134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4422..4443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4458..4488
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4496..4512
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4520..4536
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4555..4573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"