LOCUS       Exported                4217 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNW33N, complete sequence.
ACCESSION   AY237122
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4217)
  AUTHORS   Mee EK, Welker NE.
  TITLE     Development of Genetic Tools to Analyze and Characterize Genes of 
            the Thermophile Geobacillus stearothermophilus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4217)
  AUTHORS   Mee EK, Welker NE.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2003) Biochemistry, Molecular Biology, and 
            Cellular Biology, Northwestern University, 2205 Tech Drive, 
            Evanston, IL 60208, USA
REFERENCE   3  (bases 1 to 4217)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4217
                     /organism="Cloning vector pNW33N"
                     /mol_type="genomic DNA"
                     /note="shuttle vector for Escherichia coli, Bacillus 
                     subtilis, and Geobacillus stearothermophilus"
                     /db_xref="taxon:226614"
     misc_feature    1..51
                     /label=multiple cloning site from plasmid pUC19
                     /note="multiple cloning site from plasmid pUC19"
     primer_bind     complement(69..85)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    93..109
                     /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(117..147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(313..963)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label=cat
                     /note="derived from rom Staphylococcus aureus plasmid 
                     pC194"
                     /protein_id="AAO86060.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            complement(313..963)
                     /gene="cat"
                     /label=cat
     CDS             complement(1070..2074)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /note="replication protein; rolling circle replication 
                     initiation protein; Bacillus origin of replication from 
                     Geobacillus stearothermophilus plasmid pTHT15"
                     /protein_id="AAO86061.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(1070..2074)
                     /gene="repB"
                     /label=repB
     rep_origin      complement(2766..3354)
                     /direction=LEFT
                     /note="Escherichia coli origin of replication from plasmid 
                     pUC19"
     misc_feature    4217..56
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"