LOCUS       Exported                6805 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLS1ROM plasmid pLS1ROM, complete sequence.
ACCESSION   JN381945
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6805)
  AUTHORS   Ruiz-Maso JA, Lopez-Aguilar C, Nieto C, Sanz M, Buron P, Espinosa M,
            del Solar G.
  TITLE     Construction of a plasmid vector based on the pMV158 replicon for 
            cloning and inducible gene expression in Streptococcus pneumoniae
  JOURNAL   Plasmid 67 (1), 53-59 (2012)
  PUBMED    21946126
REFERENCE   2  (bases 1 to 6805)
  AUTHORS   Ruiz-Maso JA, Lopez-Aguilar C, Nieto C, Sanz M, Buron P, Espinosa M,
            Del Solar G.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUL-2011) Molecular Microbiology and Infection 
            Biology, Centro de Investigaciones Biologicas (CSIC), Ramiro de 
            Maeztu 9, Madrid, Madrid 28040, Spain
REFERENCE   3  (bases 1 to 6805)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6805
                     /organism="Cloning vector pLS1ROM"
                     /plasmid="pLS1ROM"
                     /mol_type="other DNA"
                     /label=derived from Streptococcus agalactiae
                     /note="derived from Streptococcus agalactiae"
                     /db_xref="taxon:1092445"
     rep_origin      401..570
                     /note="double strand origin; pMV158 replicon"
     CDS             655..792
                     /codon_start=1
                     /product="CopG"
                     /label=CopG
                     /note="copy number control element; transcriptional 
                     repressor protein"
                     /protein_id="AEP83061.1"
                     /translation="MKKRLTITLSESVLENLEKMAREMGLSKSAMISVALENYKKGQEK
                     "
     CDS             853..1485
                     /codon_start=1
                     /product="RepB"
                     /label=RepB
                     /note="replication initiator protein"
                     /protein_id="AEP83062.1"
                     /translation="MAKEKARYFTFLLYPESIPSDWELKLETLGVPMAISPLHDKDKSS
                     IKGQKYKKAHYHVLYIAKNPVTADSVRKKIKLLLGEKSLAMVQVVLNVENMYLYLTHES
                     KDAIAKKKHVYDKADIKLINNFDIDRYVTLDVEEKTELFNVVVSLIRAYTLQNIFDLYD
                     FIDENGETYGLTINLVNEVIAGKTGFMKLLFDGAYQRSKRGTKNEER"
     misc_feature    1583..1610
                     /label=promoter Tet coupled to malR gene
                     /note="promoter Tet coupled to malR gene"
     CDS             1986..2972
                     /codon_start=1
                     /product="MalR"
                     /label=MalR
                     /note="maltosaccharide regulatorl; transcriptional 
                     repressor protein"
                     /protein_id="AEP83063.1"
                     /translation="MPVTIKDVAKAAGVSPSTVTRVIQNKSTISDETKKRVRKAMKELN
                     YHPNLNARSLVSSYTQVIGLVLPDDSDAFYQNPFFPSVLRGISQVASENHYAIQIATGK
                     DEKERLNAISQMVYGKRVDGLIFLYAQEEDPLVKLVAEEQFPFLILGKSLSPFIPLVDN
                     DNVQAGFDATEYFIKKGCKRIAFIGGSKKLFVTKDRLTGYEQALKHYKLTTDNNRIYFA
                     DEFLEEKGYKFSKRLFKHDPQIDAIITTDSLLAEGVCNYIAKHQLDVPVLSFDSVNPKL
                     NLAAYVDINSLELGRVSLETILQIINDNKNNKQICYRQLIAHKIIEK"
     primer_bind     complement(3119..3135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3143..3159
                     /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(3167..3197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3212..3233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             4255..4992
                     /codon_start=1
                     /product="ErmAM"
                     /label=ErmAM
                     /note="adenine methylase; erythromycin resistance"
                     /protein_id="AEP83064.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     regulatory      5236..5276
                     /regulatory_class="terminator"
                     /note="tetL transcriptional terminator"
     misc_feature    5564..5605
                     /label=multi-cloning site
                     /note="multi-cloning site"
     misc_feature    complement(5655..5683)
                     /label=promoter M of the malMP operon
                     /note="promoter M of the malMP operon"
     RBS             5802..5810
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     rep_origin      6419..6628
                     /label=ssoA type of lagging-strand origin
                     /note="ssoA type of lagging-strand origin"