LOCUS       Exported               10621 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pK18msr DNA, complete sequence.
ACCESSION   AB694752
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10621)
  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 10621)
  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 10621)
  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   4  (bases 1 to 10621)
  AUTHORS   Yamamoto S, Suzuki K.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JAN-2012) Contact:Shinji Yamamoto Hiroshima 
            University, Graduate School of Science; Kagamiyama 1-3-1, 
            Higashi-Hiroshima, Hiroshima 739-8526, Japan
REFERENCE   5  (bases 1 to 10621)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10621
                     /organism="Cloning vector pK18msr"
                     /mol_type="other DNA"
                     /note="pK18msr is a shuttle vector between Escherichia coli
                     and Agrobacterium species"
                     /db_xref="taxon:1136745"
     CDS             346..1140
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin-kanamycin phosphotransferase"
                     /label=nptII
                     /protein_id="BAL46513.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            346..1140
                     /gene="nptII"
                     /label=nptII
     promoter        1186..1631
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1632..3053
                     /codon_start=1
                     /gene="sacB"
                     /product="levan sucrase"
                     /label=sacB
                     /protein_id="BAL46514.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            1632..3053
                     /gene="sacB"
                     /label=sacB
     oriT            3407..3516
                     /note="incP origin of transfer"
     oriT            3454..3523
                     /label=RP4 oriT
                     /note="RP4 oriT"
     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..5079
                     /label=pUC oriV
                     /note="pUC oriV"
     rep_origin      4485..5073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5361..5382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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"
     misc_feature    5484..5540
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    5645..5658
                     /label=TraR binding site
                     /bound_moiety="TraR"
                     /note="Tra box"
     protein_bind    5736..5749
                     /label=TraR binding site
                     /bound_moiety="TraR"
                     /note="Tra box"
     CDS             5948..7165
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A"
                     /function="involved in vegetative DNA replication"
                     /label=repA
                     /protein_id="BAL46515.1"
                     /translation="MSKAAAISRNDRPSVDVTIGEHAEQLSSQLQAMSEALFPPTSHKS
                     LRKFTSGEAARLMKISDSTLRKMTLAGEGPQPELASNGRRFYTLGQINEIRQMLAGSTR
                     GRESIDFVPHRRGSEHLQVVAVTNFKGGSGKTTTSAHLAQYLALQGYRVLAVDLDPQAS
                     LSALLGVLPETDVGANETLYAAIRYDDTRRPLRDVIRPTYFDGLHLVPGNLELMEFEHT
                     TPKALTDKGTRDGLFFTRVAQAFDEVADDYDVVVIDCPPQLGFLTLSGLCAATSMVITV
                     HPQMLDIASMSQFLLMTRDLLGVVKEAGGNLQYDFIRYLLTRYEPQDAPQTKVTALLRN
                     MFEDHVLTNPMVKSAAVSDAGLTKQTLYEIGRENLTRSTYDRAMESLDAVNSEIEALIK
                     MAWGRV"
     gene            5948..7165
                     /gene="repA"
                     /label=repA
     CDS             7395..8405
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /function="involved in vegetative DNA replication"
                     /label=repB
                     /protein_id="BAL46516.1"
                     /translation="MSRKDAIDTLFLKKQPATDRAAVDKSTARVRTGAISAMGSSLQEM
                     AEGAKAAARLQDQLATGEAVVSLDPSMIDGSPIADRLPSDVDPKFEQLEASISQEGQQV
                     PVLVRPHPEAAGRYQIVYGRRRLRAAVNLRREVSAIVRNLTDCELVVAQGRENLNRADL
                     SFIEKALFALRLEDAGFDRATIIAALSTDKADLSRYITVARGIPLNLATQIGPASKAGR
                     SRWVALAEGLGKPKATDAIEAMLGSEQFKQSDSDTRFNLIFNAVSRPPAKTPKKVRAWS
                     TPKGKKAATIRQETGRTALVFDERLVPTFGEYVADQLDSLYAQFIETNGGGKLDQ"
     gene            7395..8405
                     /gene="repB"
                     /label=repB
     rep_origin      8460..8618
                     /label=pTi-SAKURA oriV
                     /note="pTi-SAKURA oriV"
     CDS             8620..9939
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C"
                     /function="involved in vegetative DNA replication"
                     /label=repC
                     /protein_id="BAL46517.1"
                     /translation="MQTHISTTSFGRRPMTLGHIASQMAAKAVASDTVAHKWQVFQHIR
                     ESRGLIGATDRSLSILNALLTFYPETALTGGAELVVWPSNEQLMARANGMPATTLRRHL
                     AILVDCGLIIRRDSPNGKRFARKGRGGEIEQAYGFDLSPIVARAEEFRDLAQTVQAEKK
                     AFRVAKERLTLLRRDIVKMIETGVEESVPGNWGRVTQTYQGIIGRLPRSAPRQLVESIG
                     QELQELCIEIRDVLESFTKTMNLDANESHIGRHKQNSNPDSKFESEYGSGKKDEAGGSV
                     AETDNVRSLPKRELPLGIVLDACPEMRELAQGGPIRHWRDLLAAAELARPMLGISPSAW
                     REARETMGEQHAAITLASIYQRAGQINNAGGYLRSLTDRAKDGKFSTWPMVMALLRAKL
                     DEQKNAVGAGKPRTAEEVEDDSRLHVSESLLKNLRKPRSW"
     gene            8620..9939
                     /gene="repC"
                     /label=repC
     primer_bind     complement(10446..10462)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"