pNMB1m_C_Nb207 vector (V015647)

LOCUS       62056_18105        7105 bp DNA     circular SYN 07-JAN-2021
DEFINITION  Yeast display vector pNMB1m_C_Nb207, complete sequence.
ACCESSION   MT543226
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  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 7105)
  AUTHORS   Steyaert J, Uchanski T, Pardon E.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAY-2020) VIB-VUB Center for Structural Biology, VIB, 
            Pleinlaan 2, Brussels 1050, Belgium
REFERENCE   3  (bases 1 to 7105)
  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 
            (31-MAY-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..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(70..573)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        610..714
                     /label=AmpR promoter
     CDS             715..1572
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1746..2334
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2658..2688
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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)."
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2940..3592
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     misc_feature    3632..3898
                     /label=appS4 leader sequence
                     /note="appS4 leader sequence"
     misc_feature    3899..3934
                     /label=Nanobody beta-strain A (residues 1-12)
                     /note="Nanobody beta-strain A (residues 1-12)"
     misc_feature    3932..3964
                     /label=multi cloning site
                     /note="multi cloning site"
     misc_feature    3962..4291
                     /label=Nanobody Nb207 C-terminal part (residues 18-128)
                     /note="Nanobody Nb207 C-terminal part (residues 18-128)"
     misc_feature    4295..4396
                     /label=linker
                     /note="linker"
     misc_feature    4397..4603
                     /label=Aga2p anchor protein
                     /note="Aga2p anchor protein"
     CDS             4625..4858
                     /label=ACP
                     /note="acyl carrier protein (ACP) from E. coli"
     misc_feature    4862..4891
                     /label=Myc-tag
                     /note="Myc-tag"
     regulatory      4905..5211
                     /label=MFa1
                     /note="MFa1"
                     /regulatory_class="terminator"
     promoter        complement(5226..5244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5251..5267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5412..5867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(5967..6638)
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
     promoter        complement(6639..6919)
                     /label=TRP1 promoter