LOCUS       Exported                4886 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRMU824Km, complete sequence.
ACCESSION   HQ624981
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4886)
  AUTHORS   Stanislauskiene R, Gasparaviciute R, Vaitekunas J, Meskiene R, 
            Rutkiene R, Casaite V, Meskys R.
  TITLE     Construction of Escherichia coli-Arthrobacter-Rhodococcus shuttle 
            vectors based on a cryptic plasmid from Arthrobacter rhombi and 
            investigation of their application for functional screening
  JOURNAL   FEMS Microbiol. Lett. 327 (1), 78-86 (2012)
  PUBMED    22098420
REFERENCE   2  (bases 1 to 4886)
  AUTHORS   Stanislauskiene R.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-NOV-2010) Molecular Microbiology and Biotechnology, 
            Vilnius University Institute of Biochemistry, Mokslininku 12, 
            Vilnius LT-08662, Lithuania
REFERENCE   3  (bases 1 to 4886)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4886
                     /organism="Shuttle vector pRMU824Km"
                     /mol_type="other DNA"
                     /db_xref="taxon:1108803"
     protein_bind    274..295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        310..340
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    348..364
                     /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)."
     primer_bind     372..388
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             384..674
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /protein_id="AER68065.1"
                     /translation="MTMITPSSNTTHYRESLHAGLCSRRARDPISRCIRKVPSSNSLAV
                     VLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL"
     gene            384..674
                     /gene="lacZ"
                     /label=lacZ
     promoter        407..425
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(509..525)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             729..1544
                     /codon_start=1
                     /gene="kan"
                     /product="kanamycin resistance enzyme"
                     /label=kan
                     /protein_id="AER68066.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            729..1544
                     /gene="kan"
                     /label=kan
     CDS             complement(1561..1827)
                     /codon_start=1
                     /gene="repB"
                     /product="DNA-binding protein RepB"
                     /label=repB
                     /protein_id="AER68067.1"
                     /translation="MSALDPRRRKITAREAAEQVGCTPRHIRSVVAEPRHEFLARAAER
                     QRKAADWKDEGLTYREIAERLDCTPKAAENLVLRGRKARKVTA"
     gene            complement(1561..1827)
                     /gene="repB"
                     /label=repB
     CDS             complement(1827..2663)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /protein_id="AER68068.1"
                     /translation="MTAERWAEHYRQKWPMATDHDKGRFPVRQSRADALKRRYIQANPA
                     ALTTQIVIDLDHEDSLGLALELNGVPTPNYVAQSPSGHAHVAYLLAAPVCRTDNARLEP
                     MKFAARVERGLVNALRADLGYAGFMTKNPIHDGWDTVWTNDHLWTLGELATQLSGWLPR
                     SLPRRAADNSGLGRNVALFNDVRLWSYRAIRKHWEAGPDAWEQATYAYALAVNHKFAVP
                     LDASEVVHLARSVSRWTWRNFTPEGFSKVQTRRSQKAAAVRTAEKIVKAELIRSLV"
     gene            complement(1827..2663)
                     /gene="repA"
                     /label=repA
     CDS             complement(2922..3581)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl transferase"
                     /label=cat
                     /protein_id="AER68069.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(2922..3581)
                     /gene="cat"
                     /label=cat
     promoter        complement(3582..3684)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_feature    3948..4855
                     /label=replicon of plasmid p15A
                     /note="replicon of plasmid p15A"
     rep_origin      complement(4210..4755)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."