LOCUS       Exported                4513 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMESy4, complete sequence.
ACCESSION   KF415192
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4513)
  AUTHORS   Pardon E, Steyaert J.
  TITLE     A general protocol for the generation of Nanobodies for structural 
            biology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4513)
  AUTHORS   Pardon E, Steyaert J.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JUL-2013) Structural Biology Brussels, Vrije 
            Universiteit Brussel, Pleinlaan 2, Brussels 1050, Belgium
REFERENCE   3  (bases 1 to 4513)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4513
                     /organism="Cloning vector pMESy4"
                     /mol_type="other DNA"
                     /label=derived from Escherichia coli strain TG1
                     /PCR_primers="fwd_name: mp57, fwd_seq:
                     ttatgcttccggctcgtatg, rev_name: giii, rev_seq: 
                     ccacagacagccctcatag"
                     /note="derived from Escherichia coli strain TG1"
                     /db_xref="taxon:1365167"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      143..198
                     /regulatory_class="promoter"
                     /note="pLac"
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     regulatory      211..215
                     /regulatory_class="ribosome_binding_site"
     misc_feature    225..290
                     /label=encodes pelB signal peptide
                     /note="encodes pelB signal peptide"
     CDS             225..290
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     misc_feature    340..357
                     /label=His-tag
                     /note="His-tag"
     CDS             340..357
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    358..369
                     /label=CaptureSelect C-tag
                     /note="CaptureSelect C-tag"
     misc_feature    373..390
                     /label=HA-tag
                     /note="HA-tag"
     gene            403..1636
                     /gene="M13 geneIII"
                     /label=M13 geneIII
     primer_bind     complement(1642..1658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1871..2326
                     /label=f1 ori
                     /note="f1 ori"
     promoter        2608..2712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2713..3573
                     /codon_start=1
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=beta-lactamase
                     /note="ampicillin resistance"
                     /protein_id="AGT40491.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3733..4347
                     /label=pMB1
                     /note="pMB1"
     rep_origin      3744..4332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"