LOCUS       Exported                5067 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDXW-3, complete sequence.
ACCESSION   JN872903
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5067)
  AUTHORS   Tan Y, Xu D, Li Y, Wang X.
  TITLE     Construction of a novel sacB-based system for marker-free gene 
            deletion in Corynebacterium glutamicum
  JOURNAL   Plasmid 67 (1), 44-52 (2012)
  PUBMED    22100974
REFERENCE   2  (bases 1 to 5067)
  AUTHORS   Wang X, Tan Y, Xu D, Li Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2011) State Key Laboratory of Food Science and 
            Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu 
            214000, China
REFERENCE   3  (bases 1 to 5067)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5067
                     /organism="Cloning vector pDXW-3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1150128"
     terminator      26..73
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    158..203
                     /label=MCS
                     /note="MCS"
     CDS             complement(207..236)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     regulatory      237..282
                     /regulatory_class="promoter"
                     /note="PlacM"
     CDS             298..1719
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /protein_id="AFB73752.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            298..1719
                     /gene="sacB"
                     /label=sacB
     RBS             1852..1874
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2362..2553
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2655..2797
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            2984
                     /label=colE1 from pBR322
                     /note="colE1 from pBR322"
     CDS             3693..4508
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aph
                     /protein_id="AFB73751.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            3693..4508
                     /gene="aph"
                     /label=aph
     rep_origin      4601..5065
                     /label=phage f1
                     /note="phage f1"
     rep_origin      complement(4601..5056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"