LOCUS       Exported                4067 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Phagemid vector pComb3X-KD247scFv-VI, complete sequence.
ACCESSION   KP119606
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4067)
  AUTHORS   Ong YT, Kirby KA, Sarafianos SG.
  TITLE     Genetic modification of KD-247 scFv to improve affinity for the V3 
            loop of HIV-1 from multiple clades
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4067)
  AUTHORS   Ong YT, Kirby KA, Sarafianos SG.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-NOV-2014) Molecular Microbiology and Immunology, 
            University of Missouri, 401 Bond LSC, 1201 E. Rollins St., Columbia,
            MO 65211, USA
REFERENCE   3  (bases 1 to 4067)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4067
                     /organism="Phagemid vector pComb3X-KD247scFv-VI"
                     /mol_type="other DNA"
                     /db_xref="taxon:1676626"
     misc_feature    1..282
                     /label=derived from pComb3X (INSDC Accession AF268281)
                     /note="derived from pComb3X (INSDC Accession AF268281)"
     protein_bind    109..130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     gene            145..1112
                     /gene="KD-247 scFv"
                     /label=KD-247 scFv
     regulatory      145..175
                     /regulatory_class="promoter"
                     /gene="KD-247 scFv"
                     /note="lac"
     promoter        145..175
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    183..199
                     /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)."
     CDS             222..1112
                     /codon_start=1
                     /gene="KD-247 scFv"
                     /product="KD-247 single-chain variable fragment"
                     /label=KD-247 scFv
                     /note="single-chain variable fragment containing variable 
                     regions of KD-247 heavy chain and light chain"
                     /protein_id="AKQ09576.1"
                     /translation="MKKTAIAIAVALAGFATVAQAAQVQLVQSGAEVKKPGASVKVSCK
                     ASGYTFTNSWIGWFRQAPGQGLEWIGDIYPGGGYTNYNEIFKGKATMTADTSTNTAYME
                     LSSLRSEDTAVYYCSRGIPGYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQ
                     MTQRPDSLSASVGDRVTMSCKSSQSLLNSGDQKNYLTWYQQKPGQPPKLLIYWASTGES
                     GVPDRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPWTFGQGTKVEIKGQAGQHHH
                     HHHGAYPYDVPDYAS"
     sig_peptide     222..284
                     /gene="KD-247 scFv"
                     /note="ompA signal peptide"
     misc_feature    1049..4067
                     /label=derived from pComb3X (INSDC Accession AF268281)
                     /note="derived from pComb3X (INSDC Accession AF268281)"
     CDS             1056..1073
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1080..1106
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      complement(1682..2137)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2164..2268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2269..3129
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin-resistance"
                     /protein_id="AKQ09577.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3300..3888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"