LOCUS       Exported                9983 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMDS20, complete sequence.
ACCESSION   L26977
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9983)
  AUTHORS   Holmes M, Pfeifer F, Dyall-Smith M.
  TITLE     Improved shuttle vectors for Haloferax volcanii including a 
            dual-resistance plasmid
  JOURNAL   Gene 146 (1), 117-121 (1994)
  PUBMED    8063095
REFERENCE   2  (bases 1 to 9983)
  AUTHORS   Dyall-Smith ML.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-MAR-1994) Microbiology, University of Melbourne, 
            Grattan Street, Parkville, Victoria, Australia
REFERENCE   3  (bases 1 to 9983)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9983
                     /organism="Shuttle vector pMDS20"
                     /host="Haloferax alicantei Aa2.2"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:56961"
     source          894..5001
                     /organism="Shuttle vector pMDS20"
                     /mol_type="genomic DNA"
                     /note="Haloferax Aa2.2 replicon region; includes origin of 
                     replication"
                     /db_xref="taxon:56961"
     source          7680..893
                     /organism="Shuttle vector pMDS20"
                     /plasmid="Bluescript pBS+"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:56961"
     rep_origin      complement(237..692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     837..853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        860..878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5333..7252
                     /codon_start=1
                     /product="DNA gyrase beta-subunit"
                     /function="novobiocin resistance"
                     /label=DNA gyrase beta-subunit
                     /protein_id="AAB49900.1"
                     /translation="MSQDNEYGAGQIQVLEGLEAVRKRPAMYIGSTDSRGLHHLVYEVV
                     DNSIDEALAGHCDAIEVALHEDGSVSVTDNGRGIPVGTHEQYDRPALEVIMTVLHAGGK
                     FDNKSYQVSGGLHGVGVTVVNALSSELEVEVKHDGAVWTHRFEVGEPQVEEFERVRDLE
                     PGEDTGTTIRFWPDDGIFETTEFDFKTLENRLRELAFLNSGVEISLSDERTDESSTFLF
                     EGGIREFVEYLNETKTALHDDVIYYDDESEGIEVEIAMQATDELQGSIHAFANNINTRE
                     GGTHLTGFKTALTRVVNDYANSHDMLDDLDGDNLRGEDVREGLTAVISVKHPDPQFEGQ
                     TKTKLGNSEVRGIVESVTHQQLGTFFEENPDTATAIISKAVEAARARKAAKQAEELTRR
                     KSALESTSLPGKLADCQSRDPSESELFIVEGDSAGGSAKQGRDRKFQAILPLKGKILNV
                     EKHRLDRILENDEIRALITAIGGGVGDEFDIEKARYQRLILMTDADVDGAHIRTLLLTL
                     LYRHMRPLIEAGYVYAAQPPLYRVRYRGNTYDAMDEAERDRIIEEECNGNPTQVQRFKG
                     LGEMNPDQLWDTTMNPENRVLKRITVEDAAAADRMFNILMGDAVGPRKQFIKDHANDAE
                     WVDI"
     promoter        complement(7723..7741)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7762..7778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7786..7802
                     /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(7810..7840)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7855..7876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8164..8752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8923..9783)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9784..9888)
                     /gene="bla"
                     /label=AmpR promoter