LOCUS       Exported                3518 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDM276, complete sequence.
ACCESSION   EU912548
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3518)
  AUTHORS   Veltman DM, Akar G, Bosgraaf L, Van Haastert PJ.
  TITLE     A new set of small, extrachromosomal expression vectors for 
            Dictyostelium discoideum
  JOURNAL   Plasmid 61 (2), 110-118 (2009)
  PUBMED    19063918
REFERENCE   2  (bases 1 to 3518)
  AUTHORS   Veltman DM, Akar G, Bosgraaf L, van Haastert PJM.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JUL-2008) Cell Biochemistry, Rijks Universiteit 
            Groningen, Kerklaan 30, Haren 9751 NN, The Netherlands
REFERENCE   3  (bases 1 to 3518)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3518
                     /organism="Cloning vector pDM276"
                     /mol_type="other DNA"
                     /label=TAP-tag for C-terminal fusion
                     /note="TAP-tag for C-terminal fusion"
                     /db_xref="taxon:555769"
     misc_feature    10..87
                     /label=calmodulin binding peptide
                     /note="calmodulin binding peptide"
     CDS             10..87
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     misc_feature    115..135
                     /label=Tobacco Etch Virus protease recognition sequence
                     /note="Tobacco Etch Virus protease recognition sequence"
     CDS             115..135
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    175..534
                     /note="tandem repeat of Staphylococcus aureus protein A 
                     fragment"
     CDS             175..348
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             352..522
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     primer_bind     complement(579..595)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(632..650)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(671..687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    695..711
                     /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)."
     promoter        complement(719..749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    764..785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1017..1665)
                     /direction=LEFT
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      complement(1073..1661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1832..2692)
                     /codon_start=1
                     /product="penicillin beta-lactamase"
                     /label=penicillin beta-lactamase
                     /protein_id="ACH81575.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2693..2797)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2824..3279
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3420..3436
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3446..3464
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3490..3506
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"