LOCUS       40924_15590        8699 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDSK519, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8699)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1988"; volume: "70"; issue: "1"; pages: "191-197"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-NOV-2011) Biochemistry, University of Missouri, 117 Schweitzer 
            Hall, Columbia, MO 65211, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..25
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(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"
     repeat_region   631..991
                     /label=Tn903 inverted repeat
                     /note="Tn903 inverted repeat"
     CDS             1096..1908
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     repeat_region   1972..2332
                     /label=Tn903 inverted repeat
                     /note="Tn903 inverted repeat"
     rep_origin      complement(2725..3119)
                     /direction=LEFT
                     /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             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..5755
                     /codon_start=1
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             6269..7105
                     /codon_start=1
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             7095..7943
                     /codon_start=1
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     protein_bind    8610..8631
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8646..8676
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga
       61 ctctagagga tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg
      121 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      181 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      241 atggcgaatg tctggtcggt gaagtgggct ttggcgccgc caatatgttc tacgatccgg
      301 ctgacagaga cgacctttgt ctcgatccta gacgcattgc acagatggcg gacgcattct
      361 ctcgtgcgct ggacgtcgat ccgcgtcgcc tgctcgacca ggcgtacgct tatgggtgcc
      421 tttccgcagc ttggaacgcg gatggagaag aggagcaacg cgatctagct atcgcggccg
      481 cgatcaagca ggtgcgacag acgtcatact agatatcaag cgacttctcc tatcccctgg
      541 gaacacatca atctcaccgg agaatatcgc tggccaaagc cttagcgtag gattccgccc
      601 cttcccgcaa acgaccccaa acaggaaacg cagctgtaat ccgggcagcg caacggaaca
      661 ttcatcagtg taaaaatgga atcaataaag ccctgcgcag cgcgcagggt cagcctgaat
      721 acgcgtttaa tgaccagcac agtcgtgatg gcaaggtcag aatagcgctg aggtctgcct
      781 cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc cagccagaaa
      841 gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac
      901 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
      961 tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac
     1021 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1081 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctcgag gccgcgatta
     1141 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1201 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1261 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1321 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1381 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
     1441 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1501 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1561 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1621 gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact
     1681 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     1741 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     1801 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     1861 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     1921 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt
     1981 gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag
     2041 accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc
     2101 tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg
     2161 agtcagcaac accttcttca cgaggcagac ctcagcgcta ttctgacctt gccatcacga
     2221 ctgtgctggt cattaaacgc gtattcaggc tgaccctgcg cgctgcgcag ggctttattg
     2281 attccatttt tacactgatg aatgttccgt tgcgctgccc ggattacagc tgaaagcgac
     2341 caggtgctcg gcgtggcaag actcgcagcg aacccgtaga aagccatgct ccagccgccc
     2401 gcattggaga aattcttcaa attcccgttg cacatagccc ggcaattcct ttccctgctc
     2461 tgccataagc gcagcgaatg ccgggtaata ctcgtcaacg atctgataga gaagggtttg
     2521 ctcgggtcgg tggctctggt aacgaccagt atcccgatcc cggctggccg tcctggccgc
     2581 cacatgaggc atgttccgcg tccttgcaat actgtgttta catacagtct atcgcttagc
     2641 ggaaagttct tttaccctca gccgaaatgc ctgccgttgc tagacattgc cagccagtgc
     2701 ccgtcactcc cgtactaact gtcacgaacc cctgcaataa ctgtcacgcc cccctgcaat
     2761 aactgtcacg aacccctgca ataactgtca cgcccccaaa cctgcaaacc cagcaggggc
     2821 gggggctggc ggggtgttgg aaaaatccat ccatgattat ctaagaataa tccactaggc
     2881 gcggttatca gcgcccttgt ggggcgctgc tgcccttgcc caatatgccc ggccagaggc
     2941 cggatagctg gtctattcgc tgcgctaggc tacacaccgc cccaccgctg cgcggcaggg
     3001 ggaaaggcgg gcaaagcccg ctaaacccca caccaaaccc cgcagaaata cgctggagcg
     3061 cttttagccg ctttagcggc ctttccccct acccgaaggg tgggggcgcg tgtgcagccc
     3121 cgcagggcct gtctcggtcg atcattcagc ccggctcatc cttctggcgt ggcggcagac
     3181 cgaacaaggc gcggtcgtgg tcgcgttcaa ggtacgcatc cattgccgcc atgagccgat
     3241 cctccggcca ctcgctgctg ttcaccttgg ccaaaatcat ggcccccacc agcaccttgc
     3301 gccttgtttc gttcttgcgc tcttgctgct gttcccttgc ccgcacccgc tgaatttcgg
     3361 cattgattcg cgctcgttgt tcttcgagct tggccagccg atccgccgcc ttgttgctcc
     3421 ccttaaccat cttgacaccc cattgttaat gtgctgtctc gtaggctatc atggaggcac
     3481 agcggcggca atcccgaccc tactttgtag gggagggcgc acttaccggt ttctcttcga
     3541 gaaactggcc taacggccac ccttcgggcg gtgcgctctc cgagggccat tgcatggagc
     3601 cgaaaagcaa aagcaacagc gaggcagcat ggcgatttat caccttacgg cgaaaaccgg
     3661 cagcaggtcg ggcggccaat cggccagggc caaggccgac tacatccagc gcgaaggcaa
     3721 gtatgcccgc gacatggatg aagtcttgca cgccgaatcc gggcacatgc cggagttcgt
     3781 cgagcggccc gccgactact gggatgctgc cgacctgtat gaacgcgcca atgggcggct
     3841 gttcaaggag gtcgaatttg ccctgccggt cgagctgacc ctcgaccagc agaaggcgct
     3901 ggcgtccgag ttcgcccagc acctgaccgg tgccgagcgc ctgccgtata cgctggccat
     3961 ccatgccggt ggcggcgaga acccgcactg ccacctgatg atctccgagc ggatcaatga
     4021 cggcatcgag cggcccgccg ctcagtggtt caagcggtac aacggcaaga ccccggagaa
     4081 gggcggggca cagaagaccg aagcgctcaa gcccaaggca tggcttgagc agacccgcga
     4141 ggcatgggcc gaccatgcca accgggcatt agagcgggct ggccacgacg cccgcattga
     4201 ccacagaaca cttgaggcgc agggcatcga gcgcctgccc ggtgttcacc tggggccgaa
     4261 cgtggtggag atggaaggcc ggggcatccg caccgaccgg gcagacgtgg ccctgaacat
     4321 cgacaccgcc aacgcccaga tcatcgactt acaggaatac cgggaggcaa tagaccatga
     4381 acgcaatcga cagagtgaag aaatccagag gcatcaacga gttagcggag cagatcgaac
     4441 cgctggccca gagcatggcg acactggccg acgaagcccg gcaggtcatg agccagaccc
     4501 agcaggccag cgaggcgcag gcggcggagt ggctgaaagc ccagcgccag acaggggcgg
     4561 catgggtgga gctggccaaa gagttgcggg aggtagccgc cgaggtgagc agcgccgcgc
     4621 agagcgcccg gagcgcgtcg cgggggtggc actggaagct atggctaacc gtgatgctgg
     4681 cttccatgat gcctacggtg gtgctgctga tcgcatcgtt gctcttgctc gacctgacgc
     4741 cactgacaac cgaggacggc tcgatctggc tgcgcttggt ggcccgatga agaacgacag
     4801 gactttgcag gccataggcc gacagctcaa ggccatgggc tgtgagcgct tcgatatcgg
     4861 cgtcagggac gccaccaccg gccagatgat gaaccgggaa tggtcagccg ccgaagtgct
     4921 ccagaacacg ccatggctca agcggatgaa tgcccagggc aatgacgtgt atatcaggcc
     4981 cgccgagcag gagcggcatg gtctggtgct ggtggacgac ctcagcgagt ttgacctgga
     5041 tgacatgaaa gccgagggcc gggagcctgc cctggtagtg gaaaccagcc cgaagaacta
     5101 tcaggcatgg gtcaaggtgg ccgacgccgc aggcggtgaa cttcgggggc agattgcccg
     5161 gacgctggcc agcgagtacg acgccgaccc ggccagcgcc gacagccgcc actatggccg
     5221 cttggcgggc ttcaccaacc gcaaggacaa gcacaccacc cgcgccggtt atcagccgtg
     5281 ggtgctgctg cgtgaatcca agggcaagac cgccaccgct ggcccggcgc tggtgcagca
     5341 ggctggccag cagatcgagc aggcccagcg gcagcaggag aaggcccgca ggctggccag
     5401 cctcgaactg cccgagcggc agcttagccg ccaccggcgc acggcgctgg acgagtaccg
     5461 cagcgagatg gccgggctgg tcaagcgctt cggtgatgac ctcagcaagt gcgactttat
     5521 cgccgcgcag aagctggcca gccggggccg cagtgccgag gaaatcggca aggccatggc
     5581 cgaggccagc ccagcgctgg cagagcgcaa gcccggccac gaagcggatt acatcgagcg
     5641 caccgtcagc aaggtcatgg gtctgcccag cgtccagctt gcgcgggccg agctggcacg
     5701 ggcaccggca ccccgccagc gaggcatgga caggggcggg ccagatttca gcatgtagtg
     5761 cttgcgttgg tactcacgcc tgttatacta tgagtactca cgcacagaag ggggttttat
     5821 ggaatacgaa aaaagcgctt cagggtcggt ctacctgatc aaaagtgaca agggctattg
     5881 gttgcccggt ggctttggtt atacgtcaaa caaggccgag gctggccgct tttcagtcgc
     5941 tgatatggcc agccttaacc ttgacggctg caccttgtcc ttgttccgcg aagacaagcc
     6001 tttcggcccc ggcaagtttc tcggtgactg atatgaaaga ccaaaaggac aagcagaccg
     6061 gcgacctgct ggccagccct gacgctgtac gccaagcgcg atatgccgag cgcatgaagg
     6121 ccaaagggat gcgtcagcgc aagttctggc tgaccgacga cgaatacgag gcgctgcgcg
     6181 agtgcctgga agaactcaga gcggcgcagg gcgggggtag tgaccccgcc agcgcctaac
     6241 caccaactgc ctgcaaagga ggcaatcaat ggctacccat aagcctatca atattctgga
     6301 ggcgttcgca gcagcgccgc caccgctgga ctacgttttg cccaacatgg tggccggtac
     6361 ggtcggggcg ctggtgtcgc ccggtggtgc cggtaaatcc atgctggccc tgcaactggc
     6421 cgcacagatt gcaggcgggc cggatctgct ggaggtgggc gaactgccca ccggcccggt
     6481 gatctacctg cccgccgaag acccgcccac cgccattcat caccgcctgc acgcccttgg
     6541 ggcgcacctc agcgccgagg aacggcaagc cgtggctgac ggcctgctga tccagccgct
     6601 gatcggcagc ctgcccaaca tcatggcccc ggagtggttc gacggcctca agcgcgccgc
     6661 cgagggccgc cgcctgatgg tgctggacac gctgcgccgg ttccacatcg aggaagaaaa
     6721 cgccagcggc cccatggccc aggtcatcgg tcgcatggag gccatcgccg ccgataccgg
     6781 gtgctctatc gtgttcctgc accatgccag caagggcgcg gccatgatgg gcgcaggcga
     6841 ccagcagcag gccagccggg gcagctcggt actggtcgat aacatccgct ggcagtccta
     6901 cctgtcgagc atgaccagcg ccgaggccga ggaatggggt gtggacgacg accagcgccg
     6961 gttcttcgtc cgcttcggtg tgagcaaggc caactatggc gcaccgttcg ctgatcggtg
     7021 gttcaggcgg catgacggcg gggtgctcaa gcccgccgtg ctggagaggc agcgcaagag
     7081 caagggggtg ccccgtggtg aagcctaaga acaagcacag cctcagccac gtccggcacg
     7141 acccggcgca ctgtctggcc cccggcctgt tccgtgccct caagcggggc gagcgcaagc
     7201 gcagcaagct ggacgtgacg tatgactacg gcgacggcaa gcggatcgag ttcagcggcc
     7261 cggagccgct gggcgctgat gatctgcgca tcctgcaagg gctggtggcc atggctgggc
     7321 ctaatggcct agtgcttggc ccggaaccca agaccgaagg cggacggcag ctccggctgt
     7381 tcctggaacc caagtgggag gccgtcaccg ctgatgccat ggtggtcaaa ggtagctatc
     7441 gggcgctggc aaaggaaatc ggggcagagg tcgatagtgg tggggcgctc aagcacatac
     7501 aggactgcat cgagcgcctt tggaaggtat ccatcatcgc ccagaatggc cgcaagcggc
     7561 aggggtttcg gctgctgtcg gagtacgcca gcgacgaggc ggacgggcgc ctgtacgtgg
     7621 ccctgaaccc cttgatcgcg caggccgtca tgggtggcgg ccagcatgtg cgcatcagca
     7681 tggacgaggt gcgggcgctg gacagcgaaa ccgcccgcct gctgcaccag cggctgtgtg
     7741 gctggatcga ccccggcaaa accggcaagg cttccataga taccttgtgc ggctatgtct
     7801 ggccgtcaga ggccagtggt tcgaccatgc gcaagcgccg ccagcgggtg cgcgaggcgt
     7861 tgccggagct ggtcgcgctg ggctggacgg taaccgagtt cgcggcgggc aagtacgaca
     7921 tcacccggcc caaggcggca ggctgacccc ccccactcta ttgtaaacaa gacattttta
     7981 tcttttatat tcaatggctt attttcctgc taattggtaa taccatgaaa aataccatgc
     8041 tcagaaaagg cttaacaata ttttgaaaaa ttgcctactg agcgctgccg cacagctcca
     8101 taggccgctt tcctggcttt gcttccagat gtatgctctt ctttccgctt cctcgctcac
     8161 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     8221 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     8281 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     8341 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     8401 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8461 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggccc aatacgcaaa
     8521 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     8581 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8641 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataaca
//
