LOCUS       Exported                5960 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Phagemid vector pMID21, complete sequence.
ACCESSION   AY754024
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5960)
  AUTHORS   Hoet RM, Coehn EH, Kent RB, Rookey K, Schoonbroodt S, Hogan S, Rem 
            L, Frans N, Daukandt M, Pieters H, van Hegelsom R, Coolen-van Neer 
            N, Nastri HG, Rondon IJ, Leeds J, Hufton SE, Huang L, Kashin I, 
            Devlin M, Kuang G, Steukers M, Viswanathan M, Nixon AE, Sexton DJ, 
            Hoogenboom HR, Ladner RC.
  TITLE     Generation of high-affinity human antibodies by combining 
            donor-derived and synthetic complementarity-determining-region 
            diversity
  JOURNAL   Nat. Biotechnol. 23 (3), 344-348 (2005)
  PUBMED    15723048
REFERENCE   2  (bases 1 to 5960)
  AUTHORS   Ladner RC, Hoogenboom HR, Hoet RM, Cohen EH, Kashin I, Rondon IJ, 
            Rem L, Frans N, Schoonbroodt S, Kent RB, Rookey K, Hogan S.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-SEP-2004) Research, Dyax Corp, 300 Technology Square, 
            Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 5960)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5960
                     /organism="Phagemid vector pMID21"
                     /mol_type="other DNA"
                     /note="derived from M13mp18 phage cloning vector in GenBank
                     Accession Number M77815; has high-affinity synthetic and 
                     donor-derived diversity"
                     /db_xref="taxon:296377"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ApR"
                     /protein_id="AAV54523.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     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"
     misc_feature    2269..2325
                     /note="encodes light chain signal sequence; antibody 
                     stuffer"
     misc_feature    2326..2352
                     /label=encodes light chain antibody stuffer
                     /note="encodes light chain antibody stuffer"
     misc_feature    2726..2788
                     /note="encodes heavy chain signal sequence; antibody 
                     stuffer"
     misc_feature    2867..3766
                     /label=encodes heavy chain antibody stuffer
                     /note="encodes heavy chain antibody stuffer"
     misc_feature    4516..4533
                     /note="encodes hexahistidine tag; hexa-His tag"
     CDS             4516..4533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4534..4584
                     /label=encodes c-Myc tag
                     /note="encodes c-Myc tag"
     CDS             4543..4572
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    4612..4983
                     /note="encodes domain 3 of protein III; antibody stuffer"
     primer_bind     complement(5092..5108)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(5304..5759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"