LOCUS       Exported                5719 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pK18mobsacB, complete sequence.
ACCESSION   FJ437239
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5719)
  AUTHORS   Kvitko BH, Collmer A.
  TITLE     Construction of Pseudomonas syringae pv. tomato DC3000 mutant and 
            polymutant strains
  JOURNAL   Methods Mol. Biol. 712, 109-128 (2011)
  PUBMED    21359804
REFERENCE   2  (bases 1 to 5719)
  AUTHORS   Kvitko BH, Collmer A.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-NOV-2008) Plant Pathology and Plant-Microbe Biology, 
            Cornell University, 334 Plant Science, Ithaca, NY 14853, USA
REFERENCE   3  (bases 1 to 5719)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5719
                     /organism="Cloning vector pK18mobsacB"
                     /mol_type="other DNA"
                     /db_xref="taxon:576603"
     CDS             346..1140
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase"
                     /label=nptII
                     /protein_id="ACJ66844.1"
                     /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="ACJ66845.1"
                     /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=pMB1 replication origin
                     /note="pMB1 replication origin"
     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"
     primer_bind     5464..5479
                     /note="PK18M13R; special M13R binding site"
     CDS             5476..5706
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="LacZ alpha fragment"
                     /label=lacZalpha
                     /protein_id="ACJ66846.1"
                     /translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     gene            5476..5706
                     /gene="lacZalpha"
                     /label=lacZalpha
     misc_feature    5484..5540
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5544..5561)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(5544..5560)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"