pK18msr24A vector (V005267)

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LOCUS       40924_26466       12861 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Ti-curing vector pK18msr24A DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12861)
  AUTHORS   Schafer A, Tauch A, Jager W, Kalinowski J, Thierbach G, Puhler A.
  TITLE     Small mobilizable multi-purpose cloning vectors derived from the 
            Escherichia coli plasmids pK18 and pK19: selection of defined 
            deletions in the chromosome of Corynebacterium glutamicum
  JOURNAL   Gene 145 (1), 69-73 (1994)
  PUBMED    8045426
REFERENCE   2  (bases 1 to 12861)
  AUTHORS   Yamamoto S, Uraji M, Tanaka K, Moriguchi K, Suzuki K.
  TITLE     Identification of pTi-SAKURA DNA region conferring enhancement of 
            plasmid incompatibility and stability
  JOURNAL   Genes Genet. Syst. 82 (3), 197-206 (2007)
  PUBMED    17660690
REFERENCE   3  (bases 1 to 12861)
  AUTHORS   Yamamoto S, Kiyokawa K, Tanaka K, Moriguchi K, Suzuki K.
  TITLE     Novel toxin-antitoxin system composed of serine protease and 
            AAA-ATPase homologues determines the high level of stability and 
            incompatibility of the tumor-inducing plasmid pTiC58
  JOURNAL   J. Bacteriol. 191 (14), 4656-4666 (2009)
  PUBMED    19447904
REFERENCE   4  (bases 1 to 12861)
  AUTHORS   Yamamoto S, Suzuki K.
  TITLE     Development of a reinforced Ti-eviction plasmid useful for 
            construction of Ti plasmid-free Agrobacterium strains
  JOURNAL   J. Microbiol. Methods 89 (1), 53-56 (2012)
  PUBMED    22326751
REFERENCE   5  (bases 1 to 12861)
  AUTHORS   Yamamoto S, Suzuki K.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2012) Contact:Shinji Yamamoto Hiroshima 
            University, Graduate School of Science; Kagamiyama 1-3-1, 
            Higashi-Hiroshima, Hiroshima 739-8526, Japan
REFERENCE   6  (bases 1 to 12861)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 12861)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1994"; volume: "145"; issue: "1"; pages: "69-73"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Genes 
            Genet. Syst."; date: "2007"; volume: "82"; issue: "3"; pages: 
            "197-206"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "2009"; volume: "191"; issue: "14"; pages: 
            "4656-4666"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2012"; volume: "89"; issue: "1"; pages: 
            "53-56"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (20-JAN-2012) Contact:Shinji Yamamoto Hiroshima University, Graduate
            School of Science; Kagamiyama 1-3-1, Higashi-Hiroshima, Hiroshima 
            739-8526, Japan"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             346..1137
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        1186..1631
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1632..3050
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     oriT            complement(3407..3516)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        complement(3751..3841)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     promoter        complement(4035..4125)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      4485..5073
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5361..5382
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5397..5427
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5435..5451
                     /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     5459..5475
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5651..6205
                     /codon_start=1
                     /gene="tiorf24"
                     /product="putative antitoxin of toxin-antitoxin system"
                     /function="involved in plasmid stabilization"
                     /label=tiorf24
                     /protein_id="BAL46490.1"
                     /translation="MGAPLDIFKKVVERAPIKPQLVKRGGRSIRQFGQAELVFLHAYDE
                     LKLALTPKSQSEFYEALQTSLKRGLAKEVVFGKQRYDIGQHLVFVERKLKELEKLTDQV
                     DLSGKEPVIRGTHIEAHRIAALLDAGATVEEVLRDYPSLKEQQVVAARVYAEAHPKAGR
                     PYPKQTAKAAMRAADLSALDD"
     gene            5651..6205
                     /gene="tiorf24"
                     /label=tiorf24
     protein_bind    6473..6486
                     /label=TraR binding site
                     /bound_moiety="TraR"
                     /note="Tra box"
     protein_bind    6564..6577
                     /label=TraR binding site
                     /bound_moiety="TraR"
                     /note="Tra box"
     CDS             6776..7993
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A"
                     /function="involved in vegetative DNA replication"
                     /label=repA
                     /protein_id="BAL46491.1"
                     /translation="MSKAAAISRNDRPSVDVTIGEHAEQLSSQLQAMSEALFPPTSHKS
                     LRKFTSGEAARLMKISDSTLRKMTLAGEGPQPELASNGRRFYTLGQINEIRQMLAGSTR
                     GRESIDFVPHRRGSEHLQVVAVTNFKGGSGKTTTSAHLAQYLALQGYRVLAVDLDPQAS
                     LSALLGVLPETDVGANETLYAAIRYDDTRRPLRDVIRPTYFDGLHLVPGNLELMEFEHT
                     TPKALTDKGTRDGLFFTRVAQAFDEVADDYDVVVIDCPPQLGFLTLSGLCAATSMVITV
                     HPQMLDIASMSQFLLMTRDLLGVVKEAGGNLQYDFIRYLLTRYEPQDAPQTKVTALLRN
                     MFEDHVLTNPMVKSAAVSDAGLTKQTLYEIGRENLTRSTYDRAMESLDAVNSEIEALIK
                     MAWGRV"
     gene            6776..7993
                     /gene="repA"
                     /label=repA
     CDS             8223..9233
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /function="involved in vegetative DNA replication"
                     /label=repB
                     /protein_id="BAL46492.1"
                     /translation="MSRKDAIDTLFLKKQPATDRAAVDKSTARVRTGAISAMGSSLQEM
                     AEGAKAAARLQDQLATGEAVVSLDPSMIDGSPIADRLPSDVDPKFEQLEASISQEGQQV
                     PVLVRPHPEAAGRYQIVYGRRRLRAAVNLRREVSAIVRNLTDCELVVAQGRENLNRADL
                     SFIEKALFALRLEDAGFDRATIIAALSTDKADLSRYITVARGIPLNLATQIGPASKAGR
                     SRWVALAEGLGKPKATDAIEAMLGSEQFKQSDSDTRFNLIFNAVSRPPAKTPKKVRAWS
                     TPKGKKAATIRQETGRTALVFDERLVPTFGEYVADQLDSLYAQFIETNGGGKLDQ"
     gene            8223..9233
                     /gene="repB"
                     /label=repB
     rep_origin      9288..9446
                     /label=pTi-SAKURA oriV
                     /note="pTi-SAKURA oriV"
     CDS             9448..10767
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C"
                     /function="involved in vegetative DNA replication"
                     /label=repC
                     /protein_id="BAL46493.1"
                     /translation="MQTHISTTSFGRRPMTLGHIASQMAAKAVASDTVAHKWQVFQHIR
                     ESRGLIGATDRSLSILNALLTFYPETALTGGAELVVWPSNEQLMARANGMPATTLRRHL
                     AILVDCGLIIRRDSPNGKRFARKGRGGEIEQAYGFDLSPIVARAEEFRDLAQTVQAEKK
                     AFRVAKERLTLLRRDIVKMIETGVEESVPGNWGRVTQTYQGIIGRLPRSAPRQLVESIG
                     QELQELCIEIRDVLESFTKTMNLDANESHIGRHKQNSNPDSKFESEYGSGKKDEAGGSV
                     AETDNVRSLPKRELPLGIVLDACPEMRELAQGGPIRHWRDLLAAAELARPMLGISPSAW
                     REARETMGEQHAAITLASIYQRAGQINNAGGYLRSLTDRAKDGKFSTWPMVMALLRAKL
                     DEQKNAVGAGKPRTAEEVEDDSRLHVSESLLKNLRKPRSW"
     gene            9448..10767
                     /gene="repC"
                     /label=repC
     primer_bind     complement(11274..11290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             11732..12769
                     /codon_start=1
                     /gene="ietA"
                     /product="AAA-family ATPase"
                     /function="involved in plasmid stabilization"
                     /label=ietA
                     /protein_id="BAL46494.1"
                     /translation="MDHSIRRSSIYEPSSVVMTTAQQLISLLKSHVEGDDEQFLTIALQ
                     AAASEARRGHGKVAVQLRELVDAARANKDRTSGRKKAPVPIAQPRGDLANLVAVRYTDV
                     RLSSMILPAELEARLKRVILEQRQQHKLRSHGLAPRRKLLLIGPPGSGKTMTAAALAGE
                     LHTPLFTVQLDGLMTKFMGETASKLRLVFSAMAETKGVYFFDEFDAIGAKRAERNDVGE
                     IRRVLNSFLQFLEEDESDGLVVAATNHPELLDPALFRRFDDVIEYALPSVEVATGILQA
                     RLSGFDTQDIDWTKAVADAAGLSQAEIARAAADAAKLVVLSDRDRITQNDLSLAIAERK
                     GAASQ"
     gene            11732..12769
                     /gene="ietA"
                     /label=ietA