LOCUS       Exported                8699 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSK519, complete sequence.
ACCESSION   JQ173098
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8699)
  AUTHORS   Keen NT, Tamaki S, Kobayashi D, Trollinger D.
  TITLE     Improved broad-host-range plasmids for DNA cloning in gram-negative 
            bacteria
  JOURNAL   Gene 70 (1), 191-197 (1988)
  PUBMED    2853689
REFERENCE   2  (bases 1 to 8699)
  AUTHORS   Korte HL, Wall JD.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-NOV-2011) Biochemistry, University of Missouri, 117 
            Schweitzer Hall, Columbia, MO 65211, USA
REFERENCE   3  (bases 1 to 8699)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8699
                     /organism="Cloning vector pDSK519"
                     /mol_type="other DNA"
                     /db_xref="taxon:1165386"
     primer_bind     9..25
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             21..512
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGECLVGEVGFGAANMFYDPADRDDLCLDPR
                     RIAQMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY
                     "
     misc_feature    38..94
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(95..111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  631..2332
                     /mobile_element_type="transposon:Tn903"
     repeat_region   631..991
                     /label=Tn903 inverted repeat
                     /note="Tn903 inverted repeat"
     CDS             1096..1911
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AFG33237.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     repeat_region   1972..2332
                     /label=Tn903 inverted repeat
                     /note="Tn903 inverted repeat"
     repeat_region   join(2716..2753,2760..2797)
                     /rpt_family="unknown"
     rep_origin      2725..3119
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     CDS             complement(3146..3430)
                     /codon_start=1
                     /gene="mobC"
                     /product="MobC"
                     /label=mobC
                     /note="mobilization protein C"
                     /protein_id="AFG33234.1"
                     /translation="MVKGSNKAADRLAKLEEQRARINAEIQRVRAREQQQERKNETRRK
                     VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
     gene            complement(3146..3430)
                     /gene="mobC"
                     /label=mobC
     oriT            3461..3548
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             3629..5758
                     /codon_start=1
                     /gene="mobA"
                     /product="MobA"
                     /label=mobA
                     /note="mobilization protein A"
                     /protein_id="AFG33236.1"
                     /translation="MAIYHLTAKTGSRSGGQSARAKADYIQREGKYARDMDEVLHAESG
                     HMPEFVERPADYWDAADLYERANGRLFKEVEFALPVELTLDQQKALASEFAQHLTGAER
                     LPYTLAIHAGGGENPHCHLMISERINDGIERPAAQWFKRYNGKTPEKGGAQKTEALKPK
                     AWLEQTREAWADHANRALERAGHDARIDHRTLEAQGIERLPGVHLGPNVVEMEGRGIRT
                     DRADVALNIDTANAQIIDLQEYREAIDHERNRQSEEIQRHQRVSGADRTAGPEHGDTGR
                     RSPAGHEPDPAGQRGAGGGVAESPAPDRGGMGGAGQRVAGGSRRGEQRRAERPERVAGV
                     ALEAMANRDAGFHDAYGGAADRIVALARPDATDNRGRLDLAALGGPMKNDRTLQAIGRQ
                     LKAMGCERFDIGVRDATTGQMMNREWSAAEVLQNTPWLKRMNAQGNDVYIRPAEQERHG
                     LVLVDDLSEFDLDDMKAEGREPALVVETSPKNYQAWVKVADAAGGELRGQIARTLASEY
                     DADPASADSRHYGRLAGFTNRKDKHTTRAGYQPWVLLRESKGKTATAGPALVQQAGQQI
                     EQAQRQQEKARRLASLELPERQLSRHRRTALDEYRSEMAGLVKRFGDDLSKCDFIAAQK
                     LASRGRSAEEIGKAMAEASPALAERKPGHEADYIERTVSKVMGLPSVQLARAELARAPA
                     PRQRGMDRGGPDFSM"
     gene            3629..5758
                     /gene="mobA"
                     /label=mobA
     CDS             4377..4790
                     /codon_start=1
                     /gene="mobB"
                     /product="MobB"
                     /label=mobB
                     /note="mobilization protein B"
                     /protein_id="AFG33235.1"
                     /translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTQQAS
                     EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
                     MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
     gene            4377..4790
                     /gene="mobB"
                     /label=mobB
     CDS             4787..5758
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             6269..7108
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     protein_bind    8610..8631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8646..8676
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8684..1
                     /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)."