LOCUS       Exported                3595 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCL, complete sequence.
ACCESSION   AF174493
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3595)
  AUTHORS   Uhde K, Kerschbaumer RJ, Koenig R, Hirschl S, Lemaire O, Boonham N, 
            Roake W, Himmler G.
  TITLE     Improved detection of Beet necrotic yellow vein virus in a DAS ELISA
            by means of antibody single chain fragments (scFv) which were 
            selected to protease-stable epitopes from phage display libraries
  JOURNAL   Arch. Virol. 145 (1), 179-185 (2000)
  PUBMED    10664416
REFERENCE   2  (bases 1 to 3595)
  AUTHORS   Kerschbaumer RJ, Hirschl S, Himmler G.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-AUG-1999) Strain Improvement, Institute of Applied 
            Microbiology, Muthgasse 18, Vienna, AT A-1190, Austria
REFERENCE   3  (bases 1 to 3595)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3595
                     /organism="Cloning vector pCL"
                     /mol_type="genomic DNA"
                     /note="vector designed to fuse-single chain Fv to human 
                     C-kappa domain"
                     /db_xref="taxon:102687"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance gene"
                     /protein_id="AAD53875.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2274..2711
                     /codon_start=1
                     /product="PelB-IgG kappa light chain fusion protein"
                     /label=PelB-IgG kappa light chain fusion protein
                     /note="contains PelB-leader peptide, polylinker-site, 
                     constant domain from human IgG kappa light chain, and 
                     hexahistidine tag"
                     /protein_id="AAD53876.1"
                     /translation="MKYLLPTAAAGLLLLAAQPAMAQVQLQARLAAATVAAPSVFIFPP
                     SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL
                     TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGESHHHHHH"
     CDS             2274..2339
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             2691..2708
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2727..2743)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2954..3409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"