LOCUS       Exported                3714 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p34S-Km3, complete sequence.
ACCESSION   AF083408
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3714)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Plasposons: modular self-cloning minitransposon derivatives for 
            rapid genetic analysis of gram-negative bacterial genomes
  JOURNAL   Appl. Environ. Microbiol. 64 (7), 2710-2715 (1998)
  PUBMED    9647854
REFERENCE   2  (bases 1 to 3714)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Improved antibiotic-resistance cassettes through restriction site 
            elimination using Pfu DNA polymerase PCR
  JOURNAL   BioTechniques 25 (5), 772-774 (1998)
  PUBMED    9821576
REFERENCE   3  (bases 1 to 3714)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-AUG-1998) Biotechnology Center for Agriculture and the
            Environment, Cook College, Rutgers University, 59 Dudley Rd., New 
            Brunswick, NJ 08901, USA
REFERENCE   4  (bases 1 to 3714)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3714
                     /organism="Cloning vector p34S-Km3"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:81481"
     misc_feature    1..51
                     /label=multiple cloning site cassette
                     /note="multiple cloning site cassette"
     CDS             162..977
                     /codon_start=1
                     /product="kanamycin 3-phosphotransferase"
                     /label=kanamycin 3-phosphotransferase
                     /note="aminoglycoside phosphotransferase; aph(3)I"
                     /protein_id="AAC64939.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDEAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_difference 194
                     /replace="g"
                     /label=site-directed modification
                     /note="site-directed modification"
     misc_difference 287
                     /replace="a"
                     /label=site-directed modification
                     /note="site-directed modification"
     misc_difference 428
                     /replace="t"
                     /label=site-directed modification
                     /note="site-directed modification"
     misc_difference 440
                     /replace="t"
                     /label=site-directed modification
                     /note="site-directed modification"
     misc_difference 470
                     /replace="g"
                     /label=site-directed modification
                     /note="site-directed modification"
     misc_difference 710
                     /replace="t"
                     /label=site-directed modification
                     /note="site-directed modification"
     misc_difference 713
                     /replace="g"
                     /label=site-directed modification
                     /note="site-directed modification"
     misc_feature    978..1028
                     /label=multiple cloning site cassette
                     /note="multiple cloning site cassette"
     promoter        complement(1034..1052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1071..1087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1095..1111
                     /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)."
     promoter        complement(1119..1149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1164..1185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1473..2061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2232..3092)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAC64940.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2232..3092)
                     /gene="bla"
                     /label=bla
     promoter        complement(3093..3197)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3671..3687
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3690..3708
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"