LOCUS       Exported                9602 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pABC1-loxB, complete sequence.
ACCESSION   KY031730
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9602)
  AUTHORS   Doehlemann J, Wagner M.
  TITLE     A family of single copy repABC-type shuttle vectors stably 
            maintained in the alpha-proteobacterium Sinorhizobium meliloti
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9602)
  AUTHORS   Doehlemann J, Wagner M.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-OCT-2016) LOEWE-Zentrum fuer Synthetische 
            Mikrobiologie, Philipps-Universitaet Marburg, Hans-Meerwein-Strasse,
            Marburg, Hesse 35043, Germany
REFERENCE   3  (bases 1 to 9602)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9602
                     /organism="Cloning vector pABC1-loxB"
                     /mol_type="other DNA"
                     /db_xref="taxon:1928847"
     misc_feature    1..4558
                     /label=oriVSm part (oriV_p42b) derived from pLoriV-42b
                     /note="oriVSm part (oriV_p42b) derived from pLoriV-42b"
     misc_feature    4559..5278
                     /note="synTer-MCS part (synTer1-loxL) derived from 
                     pLsynTer-1-lox"
     terminator      4614..4641
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      4779..4806
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      4943..5029
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    5113..5142
                     /label=MCS
                     /note="MCS"
     misc_feature    5143..5176
                     /label=loxL
                     /note="loxL"
     protein_bind    5143..5176
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     terminator      5208..5235
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    5279..5964
                     /label=oriVEc part (oriV_pMB1) derived from pK18mob2
                     /note="oriVEc part (oriV_pMB1) derived from pK18mob2"
     rep_origin      5328..5916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5965..8663
                     /note="oriT part (mob-loxR-rrnBT2) derived from 
                     pLoriT-1-loxT"
     promoter        6058..6503
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             6504..7925
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     oriT            8279..8388
                     /note="incP origin of transfer"
     misc_feature    8531..8564
                     /label=loxR
                     /note="loxR"
     protein_bind    8531..8564
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     terminator      8590..8617
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    8664..9602
                     /note="AR part (spectinomycin resistance cassette) derived 
                     from pLAR-Spec"
     CDS             complement(8705..9496)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"