LOCUS       Exported                3306 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBEN66 DNA, complete sequence.
ACCESSION   D85525
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3306)
  AUTHORS   Yamamoto Y, Furuyama J.
  TITLE     One-step disruption by circular DNA in Escherichia coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3306)
  AUTHORS   Yamamoto Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAY-1996) Contact:Yoshihiro Yamamoto Hyogo College of 
            Medicine, Department of Genetics; Mukogawa-cho 1-1, Nishinomiya, 
            Hyogo 663, Japan
REFERENCE   3  (bases 1 to 3306)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3306
                     /organism="Cloning vector pBEN66"
                     /plasmid="pBEN66"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:47800"
     promoter        17..35
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      19..36
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     primer_bind     65..81
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(115..131)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             260..1075
                     /codon_start=1
                     /gene="kan"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=kan
                     /note="kanamycin resistance gene from Tn903"
                     /protein_id="BAA12824.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            260..1075
                     /gene="kan"
                     /label=kan
     rep_origin      complement(1450..2038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2209..3069)
                     /codon_start=1
                     /gene="amp"
                     /product="beta-lactamase"
                     /label=amp
                     /note="amipicillin resistance gene"
                     /protein_id="BAA12825.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2209..3069)
                     /gene="amp"
                     /label=amp
     promoter        complement(3070..3174)
                     /gene="bla"
                     /label=AmpR promoter