LOCUS       Exported               12689 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression shuttle vector pWLG30, complete sequence.
ACCESSION   AF134199
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12689)
  AUTHORS   Gardner WL, Whitman WB.
  TITLE     Expression vectors for Methanococcus maripaludis: overexpression of 
            acetohydroxyacid synthase and beta-galactosidase
  JOURNAL   Genetics 152 (4), 1439-1447 (1999)
  PUBMED    10430574
REFERENCE   2  (bases 1 to 12689)
  AUTHORS   Gardner WL, Whitman WB.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-MAR-1999) Microbiology, University of Georgia, 
            Biological Sciences Building, Athens, GA 30602, USA
REFERENCE   3  (bases 1 to 12689)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12689
                     /organism="Expression shuttle vector pWLG30"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:100613"
     regulatory      61..69
                     /regulatory_class="promoter"
                     /note="PhmvA BoxA; Methanococcus voltae histone promoter 
                     Box A"
     regulatory      92..95
                     /regulatory_class="promoter"
                     /note="PhmvA BoxB; Methanococcus voltae histone promoter 
                     Box B"
     regulatory      153..158
                     /regulatory_class="ribosome_binding_site"
     misc_feature    160..205
                     /label=polylinker
                     /note="polylinker"
     regulatory      464..471
                     /regulatory_class="promoter"
                     /note="Pmcr Box A; promoter for Methanococcus voltae methyl
                     coenzyme M reductase"
     CDS             988..1587
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin transacetylase"
                     /label=pac
                     /note="from Streptomyces alboniger"
                     /protein_id="AAF01148.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTRAARTLPHARRARHRQGVGRATRRRGGGLDHAGERRSGGGVRRDRNAHGRVERFPAG
                     RAATDGRPPGAAPAQGARVVPGHRRRLARPPGQGSGQRRRAPRSGGGRARRGARLPGDL
                     RAPQPPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     gene            988..1587
                     /gene="pac"
                     /label=pac
     misc_feature    1409..12689
                     /note="cryptic plasmid; pURB500 sequence from Methanococcus
                     maripaludis strain C5"
     regulatory      1887..1912
                     /regulatory_class="terminator"
                     /note="terminator for Methanococcus voltae methyl coenzyme 
                     m reductase"
     promoter        2266..2370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2371..3231
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAF01149.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2371..3231
                     /gene="bla"
                     /label=bla
     rep_origin      3402..3990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3664..3806
     protein_bind    4278..4299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4314..4344
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      4316..4344
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    4352..4368
                     /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)."
     primer_bind     4376..4392
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6816..7188
                     /label=ORFLESS2
                     /note="ORFLESS2"
     misc_feature    7948..8733
                     /label=ORFLESS1
                     /note="ORFLESS1"