LOCUS       Exported                8353 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMSP3535, complete sequence.
ACCESSION   AY303239
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8353)
  AUTHORS   Bryan EM, Bae T, Kleerebezem M, Dunny GM.
  TITLE     Improved vectors for nisin-controlled expression in gram-positive 
            bacteria
  JOURNAL   Plasmid 44 (2), 183-190 (2000)
  PUBMED    10964628
REFERENCE   2  (bases 1 to 8353)
  AUTHORS   Staddon JH, Bryan EM, Manias DA, Dunny GM.
  TITLE     Conserved target for group II intron insertion in relaxase genes of 
            conjugative elements of gram-positive bacteria
  JOURNAL   J. Bacteriol. 186 (8), 2393-2401 (2004)
  PUBMED    15060042
REFERENCE   3  (bases 1 to 8353)
  AUTHORS   Staddon JH, Bryan EM, Manias DA, Bae T, Kleerebezem M, Dunny GM.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAY-2003) Microbiology, University of Minnesota, MMC 
            196, 420 Delaware Street S.E., Minneapolis, MN 55455, USA
REFERENCE   4  (bases 1 to 8353)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8353
                     /organism="Cloning vector pMSP3535"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:233882"
     CDS             332..1069
                     /codon_start=1
                     /product="Erm"
                     /label=Erm
                     /note="erythromycin resistance protein; adenine methylase"
                     /protein_id="AAP60029.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     CDS             2046..2345
                     /codon_start=1
                     /product="RepD"
                     /label=RepD
                     /protein_id="AAP60030.1"
                     /translation="MNHDECKTYIKNSLLEIRKLANIYTLETFKKELEKRNIYLETKSD
                     KYFSSEGEDYIYKLIENNKIIYSISGKKLTYKGKKSFSKHAILKQLNEKANQVN"
     CDS             2388..3878
                     /codon_start=1
                     /product="RepE"
                     /label=RepE
                     /note="plasmid replication; pAMbeta1 plasmid replicon"
                     /protein_id="AAP60031.1"
                     /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
                     SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
                     FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
                     VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
                     QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
                     SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
                     DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
                     VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
                     ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
     CDS             4228..4365
                     /codon_start=1
                     /product="RepG"
                     /label=RepG
                     /protein_id="AAP60032.1"
                     /translation="MEKYLKNPITWIGLVLVVTWFLTKSSEFLIFGVCVLLLVFASQSD
                     "
     CDS             4614..5252
                     /codon_start=1
                     /product="NisR"
                     /label=NisR
                     /note="response regulator"
                     /protein_id="AAP60033.1"
                     /translation="MKTALEMRNYEVATHQNISLPLDITDFQGFDLILLDIMMSNIEGT
                     EICKRIRREISTPIIFVSAKDTEEDIINGLGIGGDDYITKPFSLKQLVAKVEANIKREE
                     RNKHAVHVFSEIRRDLGPITFYLEERRVCVNGQTIPLTCREYDILELLSQRTSKVYTRE
                     DIYDDVYDEYSNALFRSISEYIYQIRSKFAPYDINPIKTVRGLGYQWHG"
     CDS             5245..6588
                     /codon_start=1
                     /product="NisK"
                     /label=NisK
                     /note="histidine kinase"
                     /protein_id="AAP60034.1"
                     /translation="MGKKYSMRRRIWQAVIEIIIGTCLLILLLLGLTFFLRQIGQISGS
                     ETIRLSLDSDNLTISDIERDMKHYPYDYIIFDNDTSKILGGHYVKSDVPSFVASKQSSH
                     NITEGEITYTYSSNKHFSVVLRQNSMPEFTNHTLRSISYNQFTYLFFFLGEIILIIFSV
                     YHLIREFSKNFQAVQKIALKMGEITTFPEQEESKIIEFDQVLNNLYSKSKELAFLIEAE
                     RHEKHDLSFQVAALSHDVKTPLTVLKGNIELLEMTEVNEQQADFIESMKNSLTVFDKYF
                     NTMISYTKLLNDENDYKATISLEDFLIDLSVELEELSTTYQVDYQLVKKTDLTTFYGNT
                     LALSRALINIFVNACQYAKEGEKIVSLSIYDDEKYLYFEIWNNGHPFSEQAKKNAGKLF
                     FTEDTGRSGKHYGIGLSFAQGVALKHQGNLILSNPQKGGAEVILKIKK"
     rep_origin      6624..7969
                     /note="ColE1 ori; cloned from pSP73"
     rep_origin      7090..7678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        7936..7954
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      8045..8353
                     /regulatory_class="promoter"
                     /note="PnisA; nisin inducible promoter"