LOCUS       Exported                6134 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pLES003 DNA, complete sequence.
ACCESSION   AB370338
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6134)
  AUTHORS   Wada T, Noda M, Kashiwabara F, Jeon HJ, Shirakawa A, Yabu H, Matoba 
            Y, Kumagai T, Sugiyama M.
  TITLE     Characterization of four plasmids harboured in a Lactobacillus 
            brevis strain encoding a novel bacteriocin, brevicin 925A, and 
            construction of a shuttle vector for lactic acid bacteria and 
            Escherichia coli
  JOURNAL   Microbiology (Reading, Engl.) 155 (PT 5), 1726-1737 (2009)
  PUBMED    19372160
REFERENCE   2  (bases 1 to 6134)
  AUTHORS   Noda M, Sugiyama M.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2007) Contact:Masafumi Noda Hiroshima University, 
            Graduate School of Biomedical Sciences; Kasumi 1-2-3, Minami-ku, 
            Hiroshima 734-8553, Japan
REFERENCE   3  (bases 1 to 6134)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6134
                     /organism="Shuttle vector pLES003"
                     /mol_type="other DNA"
                     /label=shuttle vector pLES003 from L. brevis and E. coli
                     /note="shuttle vector pLES003 from L. brevis and E. coli"
                     /db_xref="taxon:487657"
     CDS             299..1036
                     /codon_start=1
                     /gene="erm"
                     /product="MLS resistance protein"
                     /label=erm
                     /note="macrolide-lincosamide-streptogramin B (MLS) 
                     resistance"
                     /protein_id="BAH56452.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     gene            299..1036
                     /gene="erm"
                     /label=erm
     misc_feature    complement(1343..1666)
                     /label=LacZ alpha CDS
                     /note="LacZ alpha CDS"
     CDS             complement(1379..1666)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRI"
     primer_bind     1576..1592
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1593..1649
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    1593..1637
                     /note="multi cloning site 
                     (EcoRI-SacI-KpnI-SmaI-BamHI-XbaI-SalI-PstI); poly linker"
     primer_bind     complement(1662..1678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1686..1702
                     /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(1710..1740)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1755..1776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2064..2652)
                     /direction=LEFT
                     /note="replication origin in E. coli (ColE1 origin from 
                     pUC19)"
     CDS             complement(2823..3683)
                     /codon_start=1
                     /gene="amp"
                     /product="beta-lactamase"
                     /label=amp
                     /note="ampicillin resistance protein"
                     /protein_id="BAH56453.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2823..3683)
                     /gene="amp"
                     /label=amp
     promoter        complement(3684..3788)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4357..5280)
                     /codon_start=1
                     /gene="repA"
                     /product="plasmid replication protein"
                     /label=repA
                     /protein_id="BAH56454.1"
                     /translation="MSNELVKYQPELNTIPLRKFSPTEMNLFFSIISRMRDKGDQTVRF
                     SFDQLKDLSNYKPTANRRFIDDLKRTYNHLMDLRFGSQSKSGLSFERFVMFTKFKINGD
                     VEEPYVDIEIYKDALPLLNHLESWVRYALVEFRDLKSSYAKTMFRLLKGYRTTGYAYFS
                     KSDFIELLDIPKSYRQSNTDQYVISPIKEELTPLFRGLTVRKKYGKGRGKPVIGYSFAW
                     KPERKDAEDVHVSKTERLKKAKFNIEHNGELSDKEKWRAIDKIKGLKLGTTEAEHNKQE
                     QEAREEQIRAEERQKILEKLRNGGLR"
     gene            complement(4357..5280)
                     /gene="repA"
                     /label=repA
     rep_origin      complement(5336..5509)
                     /direction=LEFT
                     /note="replication origin in Gram-positive bacteria (iteron
                     region)"