LOCUS       Exported                7828 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pELS200, complete sequence.
ACCESSION   AY557621
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7828)
  AUTHORS   Diep DB, Johnsborg O, Risoen PA, Nes IF.
  TITLE     Evidence for dual functionality of the operon plnABCD in the 
            regulation of the operon plnABCD in the regulation of bacteriocin 
            production in Lactobacillus plantarum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7828)
  AUTHORS   Skaugen M, Axelsson L.
  TITLE     pELS200 E.coli/LActibacillus shuttle vector
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7828)
  AUTHORS   Skaugen M, Axelsson L.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-FEB-2004) Dept. of Chemistry, Biotechnology, and Food 
            Science, Agricultural University of Norway, Chr. Magnus Falsens vei 
            1, Aas N-1432, Norway
REFERENCE   4  (bases 1 to 7828)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7828
                     /organism="Shuttle vector pELS200"
                     /mol_type="other DNA"
                     /note="derived from E. coli plasmids RK2, pGEM7-Zf(+), 
                     pBluescript SKII, and Lactobacillus plasmids p256 and 
                     pLUL631; E.coli/Lactobacillus shuttle vector"
                     /db_xref="taxon:268235"
     CDS             190..441
                     /codon_start=1
                     /gene="parD"
                     /product="ParD"
                     /label=parD
                     /note="segregational stability determinant; RK2 origin"
                     /protein_id="AAS77681.1"
                     /translation="MSRLTIDMTDQQHQSLKALAALQGKTIKQYALERLFPGDADADQA
                     WQELKTMLGNRINDGLAGKVSTKSVGEILDEELSGDRA"
     gene            190..441
                     /gene="parD"
                     /label=parD
     CDS             438..749
                     /codon_start=1
                     /gene="parE"
                     /product="ParE"
                     /label=parE
                     /note="segregational stability determinant; RK2 origin"
                     /protein_id="AAS77682.1"
                     /translation="MTAYILTAEAEADLRGIIRYTRREWGAAQVRRYIAKLEQGIARLA
                     AGEGPFKDMSELFPALRMARCEHHYVFCLPRAGEPALVVAILHERMDLMTRLADRLKG"
     gene            438..749
                     /gene="parE"
                     /label=parE
     primer_bind     883..899
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1063..1800)
                     /codon_start=1
                     /gene="ermB"
                     /product="ErmB"
                     /label=ermB
                     /note="23S rRNA methylase"
                     /protein_id="AAS77683.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGNIPYN
                     LSTQIIKKVVFESRASDIYLTVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYKYFVSKWVNREYRQLFTKNQFRQAMKHAKVNN
                     LSTITYEQVLSIFNSYLLFNGRK"
     gene            complement(1063..1800)
                     /gene="ermB"
                     /label=ermB
     CDS             complement(1897..1980)
                     /codon_start=1
                     /gene="ermBL"
                     /product="ErmBL"
                     /label=ermBL
                     /note="regulatory protein"
                     /protein_id="AAS77684.1"
                     /translation="MLVFQMRNVDKTSTVLKQTKNSDYADK"
     gene            complement(1897..1980)
                     /gene="ermBL"
                     /label=ermBL
     oriT            3275..3383
                     /note="incP origin of transfer"
     promoter        3560..3664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3665..4525
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     mRNA            3665..4522
     rep_origin      4772..5302
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4871..5296
                     /note="oriV; RK2 vegetative replication origin"
     regulatory      5524..5642
                     /regulatory_class="promoter"
                     /note="Pneo; neomycin phosphotransferase promoter"
     CDS             5694..6842
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /label=trfA
                     /note="RK2 replication initiator"
                     /protein_id="AAS77685.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            5694..6842
                     /gene="trfA"
                     /label=trfA
     CDS             complement(7107..7487)
                     /codon_start=1
                     /gene="lacZ-alpha"
                     /product="LacZ-alpha"
                     /label=lacZ-alpha
                     /note="chimaeric (pGEM/pBluescript) alpha peptide; allows 
                     blue/white screening"
                     /protein_id="AAS77686.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     gene            complement(7107..7487)
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     primer_bind     7271..7287
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7297..7315
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7324..7431
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     misc_feature    7330..7431
                     /label=multicloning site
                     /note="multicloning site"
     primer_bind     7341..7357
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7391..7407)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7444..7462)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7483..7499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7507..7523
                     /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(7531..7561)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7576..7597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."