pMESD2 vector (V016039)

LOCUS       62056_16745        5754 bp DNA     circular SYN 07-JAN-2021
DEFINITION  Cloning vector pMESD2, complete sequence.
ACCESSION   MT328400
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5754)
  AUTHORS   Uchanski T, Masiulis S, Fischer B, Kalichuk V, Lopez-Sanchez U, 
            Zarkadas E, Weckener M, Sente A, Ward P, Wohlkoenig A, Zoegg T, 
            Remaut H, Naismith JH, Nury H, Vranken W, Aricescu AR, Pardon E, 
            Steyaert J.
  TITLE     Megabodies expand the nanobody toolkit for protein structure 
            determination by single-particle cryo-EM
  JOURNAL   Nat Methods 18 (1), 60-68 (2021)
REFERENCE   2  (bases 1 to 5754)
  AUTHORS   Steyaert J, Uchanski T, Pardon E.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-APR-2020) VIB-VUB Center for Structural Biology, VIB, 
            Pleinlaan 2, Brussels 1050, Belgium
REFERENCE   3  (bases 1 to 5754)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods"; date: "2021"; volume: "18"; issue: "1"; pages: "60-68"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-APR-2020) VIB-VUB Center for Structural Biology, VIB, Pleinlaan 
            2, Brussels 1050, Belgium"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /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)."
     misc_feature    225..281
                     /label=DsbA signal peptide
                     /note="DsbA signal peptide"
     misc_feature    282..320
                     /label=nanobody beta-strain A (residues 1-14)
                     /note="nanobody beta-strain A (residues 1-14)"
     misc_feature    321..1544
                     /label=cHopQ scaffold protein
                     /note="cHopQ scaffold protein"
     misc_feature    1545..1547
                     /label=nanobody residue 17
                     /note="nanobody residue 17"
     misc_feature    1548..1580
                     /label=multi cloning site
                     /note="multi cloning site"
     CDS             1581..1598
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    1599..1610
                     /label=EPEA-tag
                     /note="EPEA-tag"
     misc_feature    1614..1643
                     /label=HA-tag
                     /note="HA-tag"
     gene            1644..2877
                     /gene="M13 geneIII"
                     /label=M13 geneIII
     primer_bind     complement(2883..2899)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3112..3567
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3849..3953
                     /label=AmpR promoter
     CDS             3954..4811
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4985..5573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"