LOCUS       Exported                7022 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pELS100, complete sequence.
ACCESSION   AY557620
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7022)
  AUTHORS   Rawlinson EL, Nes IF, Skaugen M.
  TITLE     LasX, a transcriptional regulator of the lactocin S biosynthetic 
            genes in Lactobacillus sakei L45, acts both as an activator and a 
            repressor
  JOURNAL   Biochimie 84 (5-6), 559-567 (2002)
  PUBMED    12423800
REFERENCE   2  (bases 1 to 7022)
  AUTHORS   Sorvig E, Skaugen M, Naterstad K, Eijsink VG, Axelsson L.
  TITLE     Plasmid p256 from Lactobacillus plantarum represents a new type of 
            replicon in lactic acid bacteria, and contains a 
            toxin-antitoxin-like plasmid maintenance system
  JOURNAL   Microbiology (Reading, Engl.) 151 (PT 2), 421-431 (2005)
  PUBMED    15699191
REFERENCE   3  (bases 1 to 7022)
  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 7022)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7022
                     /organism="Shuttle vector pELS100"
                     /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:268234"
     primer_bind     77..93
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(257..994)
                     /codon_start=1
                     /gene="ermB"
                     /product="ErmB"
                     /label=ermB
                     /note="confers erythromycin resistance; 23S rRNA methylase"
                     /protein_id="AAS77676.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGNIPYN
                     LSTQIIKKVVFESRASDIYLTVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYKYFVSKWVNREYRQLFTKNQFRQAMKHAKVNN
                     LSTITYEQVLSIFNSYLLFNGRK"
     gene            complement(257..994)
                     /gene="ermB"
                     /label=ermB
     CDS             complement(1091..1174)
                     /codon_start=1
                     /gene="ermBL"
                     /product="ErmBL"
                     /label=ermBL
                     /note="regulatory protein"
                     /protein_id="AAS77677.1"
                     /translation="MLVFQMRNVDKTSTVLKQTKNSDYADK"
     gene            complement(1091..1174)
                     /gene="ermBL"
                     /label=ermBL
     misc_feature    1315..2217
                     /note="p256 replication region; allows replication in 
                     Lactobacillus species"
     misc_feature    2469..2579
                     /note="oriT; RK2 origin of transfer"
     oriT            2469..2577
                     /note="incP origin of transfer"
     promoter        2754..2858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2859..3719
                     /codon_start=1
                     /gene="bla"
                     /product="Bla beta lactamase"
                     /label=bla
                     /protein_id="AAS77678.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2859..3719
                     /gene="bla"
                     /label=bla
     rep_origin      3966..4496
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4065..4490
                     /note="oriV; RK2 vegetative replication origin"
     regulatory      4718..4836
                     /regulatory_class="promoter"
                     /note="Pneo; neomycin phosphotransferase promoter"
     CDS             4888..6036
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /label=trfA
                     /note="essential for replication; RK2 replication 
                     initiator"
                     /protein_id="AAS77679.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            4888..6036
                     /gene="trfA"
                     /label=trfA
     CDS             complement(6301..6681)
                     /codon_start=1
                     /gene="lacZ-alpha"
                     /product="LacZ-alpha"
                     /label=lacZ-alpha
                     /note="chimaeric (pGEM/pBluescript) alpha peptide; allows 
                     blue/white screening"
                     /protein_id="AAS77680.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     gene            complement(6301..6681)
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     primer_bind     6465..6481
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6491..6509
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6518..6625
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     misc_feature    6524..6625
                     /label=multicloning site
                     /note="multicloning site"
     primer_bind     6535..6551
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6585..6601)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6638..6656)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6677..6693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6701..6717
                     /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(6725..6755)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6770..6791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."