LOCUS       Exported                6811 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWKS130, complete sequence.
ACCESSION   JQ283951
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  AUTHORS   Wang RF, Kushner SR.
  TITLE     Construction of versatile low-copy-number vectors for cloning, 
            sequencing and gene expression in Escherichia coli
  JOURNAL   Gene 100, 195-199 (1991)
  PUBMED    2055470
REFERENCE   2  (bases 1 to 6811)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Low copy expression vectors for use in Yersinia sp. and related 
            organisms
  JOURNAL   Plasmid 68 (1), 33-42 (2012)
  PUBMED    22445322
REFERENCE   3  (bases 1 to 6811)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2011) Microbiology and Immunology, University of 
            North Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen 
            Jones, CB# 7290, Chapel Hill, NC 27599, USA
REFERENCE   4  (bases 1 to 6811)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: DNASTAR Lasergene SeqMan Pro v. 7.2.1 (1)
                                     Intel
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6811
                     /organism="Expression vector pWKS130"
                     /mol_type="other DNA"
                     /db_xref="taxon:1166521"
     protein_bind    240..261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    243..258
                     /label=CAP binding site
                     /bound_moiety="CAP"
                     /note="CAP binding site"
     promoter        276..306
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      276..281
                     /regulatory_class="minus_35_signal"
     regulatory      300..305
                     /regulatory_class="minus_10_signal"
     protein_bind    312..332
                     /label=lac operator binding site
                     /bound_moiety="lac operator"
                     /note="lac repressor binding site"
     protein_bind    314..330
                     /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     338..354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      339..343
                     /regulatory_class="ribosome_binding_site"
     CDS             350..925
                     /codon_start=1
                     /gene="lacZa"
                     /product="beta-galactosidase alpha peptide"
                     /label=lacZa
                     /protein_id="AFF57866.1"
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     gene            350..925
                     /gene="lacZa"
                     /label=lacZa
     promoter        375..393
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    406..513
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     423..439
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(473..489)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(522..540)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(550..566)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      708..1136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(2451..3266)
                     /codon_start=1
                     /gene="aphA1"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphA1
                     /protein_id="AFF57865.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2451..3266)
                     /gene="aphA1"
                     /label=aphA1
     misc_feature    4202..6147
                     /label=region containing pSC101 origin of replication
                     /note="region containing pSC101 origin of replication"
     CDS             complement(4603..5553)
                     /codon_start=1
                     /gene="repA"
                     /product="replication initiation protein"
                     /label=repA
                     /protein_id="AFF57864.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     gene            complement(4603..5553)
                     /gene="repA"
                     /label=repA
     rep_origin      5601..5823
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"