LOCUS       Exported                9463 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMSP3535H3, complete sequence.
ACCESSION   FJ555208
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9463)
  AUTHORS   Oddone GM, Mills DA, Block DE.
  TITLE     Incorporation of nisI-mediated nisin immunity improves vector-based 
            nisin-controlled gene expression in lactic acid bacteria
  JOURNAL   Plasmid 61 (3), 151-158 (2009)
  PUBMED    19141301
REFERENCE   2  (bases 1 to 9463)
  AUTHORS   Oddone GM, Kim J-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-DEC-2008) Viticulture and Enology, University of 
            California, Davis, RMI North Lab, 595 Hilgard Lane, Davis, CA 95616,
            USA
REFERENCE   3  (bases 1 to 9463)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9463
                     /organism="Shuttle vector pMSP3535H3"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:594678"
     CDS             332..1069
                     /codon_start=1
                     /product="ErmC"
                     /label=ErmC
                     /protein_id="ACM62691.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     CDS             1971..2270
                     /codon_start=1
                     /gene="repD"
                     /product="RepD"
                     /label=repD
                     /note="replication factor"
                     /protein_id="ACM62692.1"
                     /translation="MNHDECKTYIKNSLLEIRKLANIYTLETFKKELEKRNIYLETKSD
                     KYFSSEGEDYIYKLIENNKIIYSISGKKLTYKGKKSFSKHAILKQLNEKANQVN"
     gene            1971..2270
                     /gene="repD"
                     /label=repD
     CDS             2313..3803
                     /codon_start=1
                     /gene="repE"
                     /product="RepE"
                     /label=repE
                     /note="replication factor"
                     /protein_id="ACM62693.1"
                     /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
                     SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
                     FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
                     VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
                     QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
                     SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
                     DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
                     VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
                     ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
     gene            2313..3803
                     /gene="repE"
                     /label=repE
     CDS             4153..4290
                     /codon_start=1
                     /gene="repG"
                     /product="RepG"
                     /label=repG
                     /note="replication factor"
                     /protein_id="ACM62694.1"
                     /translation="MEKYLKNPITWIGLVLVVTWFLTKSSEFLIFGVCVLLLVFASQSD
                     "
     gene            4153..4290
                     /gene="repG"
                     /label=repG
     CDS             4440..5177
                     /codon_start=1
                     /gene="nisI"
                     /product="NisI"
                     /label=nisI
                     /note="nisin immunity protein; provides significant 
                     protection from the toxic effects of the inducer nisin"
                     /protein_id="ACM62695.1"
                     /translation="MRKYLILIVALIGITGLSGCYQTSQKKVRFDEGSYTNFIYDNKSY
                     FVTDKEIPQENVNNCKVKFYNLLIVDMKSEKLLSSSNKNSVTLVLNNIYEASDKSLCMG
                     INDRYYKILPESDKGVVKALRLQNFDVTSDISDDNFVIDKNDSRKIDYMGNIYSISDTT
                     VSDEELGEYQDVLAEVRVFDSVSGKSIPRSEWGRIDKDGSNSKQSRTEWDYGEIHSIRG
                     KSLTEAFAVEINDDFKLATKVGN"
     gene            4440..5177
                     /gene="nisI"
                     /label=nisI
     CDS             5406..6044
                     /codon_start=1
                     /gene="nisR"
                     /product="NisR"
                     /label=nisR
                     /note="nis operon regulator protein"
                     /protein_id="ACM62696.1"
                     /translation="MKTALEMRNYEVATHQNISLPLDITDFQGFDLILLDIMMSNIEGT
                     EICKRIRREISTPIIFVSAKDTEEDIINGLGIGGDDYITKPFSLKQLVAKVEANIKREE
                     RNKHAVHVFSEIRRDLGPITFYLEERRVCVNGQTIPLTCREYDILELLSQRTSKVYTRE
                     DIYDDVYDEYSNALFRSISEYIYQIRSKFAPYDINPIKTVRGLGYQWHG"
     gene            5406..6044
                     /gene="nisR"
                     /label=nisR
     CDS             6037..7380
                     /codon_start=1
                     /gene="nisK"
                     /product="NisK"
                     /label=nisK
                     /note="kinase"
                     /protein_id="ACM62697.1"
                     /translation="MGKKYSMRRRIWQAVIEIIIGTCLLILLLLGLTFFLRQIGQISGS
                     ETIRLSLDSDNLTISDIERDMKHYPYDYIIFDNDTSKILGGHYVKSDVPSFVASKQSSH
                     NITEGEITYTYSSNKHFSVVLRQNSMPEFTNHTLRSISYNQFTYLFFFLGEIILIIFSV
                     YHLIREFSKNFQAVQKIALKMGEITTFPEQEESKIIEFDQVLNNLYSKSKELAFLIEAE
                     RHEKHDLSFQVAALSHDVKTPLTVLKGNIELLEMTEVNEQQADFIESMKNSLTVFDKYF
                     NTMISYTKLLNDENDYKATISLEDFLIDLSVELEELSTTYQVDYQLVKKTDLTTFYGNT
                     LALSRALINIFVNACQYAKEGEKIVSLSIYDDEKYLYFEIWNNGHPFSEQAKKNAGKLF
                     FTEDTGRSGKHYGIGLSFAQGVALKHQGNLILSNPQKGGAEVILKIKK"
     gene            6037..7380
                     /gene="nisK"
                     /label=nisK
     rep_origin      7416..8755
                     /label=colE
                     /note="colE"
     rep_origin      7882..8470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        8728..8746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(8914..9237)
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="LacZ alpha"
                     /label=lacZalpha
                     /note="galactosidase subunit"
                     /protein_id="ACM62698.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     gene            complement(8914..9237)
                     /gene="lacZalpha"
                     /label=lacZalpha
     primer_bind     9147..9163
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9164..9220
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(9233..9249)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"