LOCUS       40924_37453        4886 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRMU824Km, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4886)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "FEMS 
            Microbiol. Lett."; date: "2012"; volume: "327"; issue: "1"; pages: 
            "78-86"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-NOV-2010) Molecular Microbiology and Biotechnology, Vilnius 
            University Institute of Biochemistry, Mokslininku 12, Vilnius 
            LT-08662, Lithuania"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    274..295
                     /label=CAP binding site
                     /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
                     /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"
     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..1541
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     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(2925..3581)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3582..3684)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     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."
ORIGIN
        1 gatcttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
       61 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
      121 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
      181 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      241 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      301 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      361 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctaat acgactcact
      421 atagggaaag cttgcatgca ggcctctgca gtcgacgggc ccgggatccg atatctagat
      481 gcattcgcaa ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
      541 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      601 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      661 aatggaaatt gtaagcctag ggaacaataa aactgtctgc ttacataaac agtaatacaa
      721 ggggtgttat gagccatatt caacgggaaa cgtcttgctc gaggccgcga ttaaattcca
      781 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
      841 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
      901 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
      961 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1021 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1081 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1141 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     1201 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1261 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1321 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1381 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1441 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1501 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagac ctaggtggcc
     1561 tcaggccgtc accttgcggg ccttgcggcc acgcagaacc aggttctcag cggccttggg
     1621 ggtgcaatcc agccgctccg caatctcccg gtaggtcagg ccctcgtcct tccagtccgc
     1681 ggctttacgc tggcgctcag ccgctctcgc caggaattca tgtcgaggct cggcaaccac
     1741 gctccggata tggcgcgggg tgcagccgac ctgctcggct gcttctcgag cggtgatctt
     1801 ccggcggcgg ggatctagtg cgctcattag acgaggctcc taatcaactc ggctttcacg
     1861 atcttttcgg cggtgcggac ggcggcagcc ttctggctcc gccgtgtctg cactttgctg
     1921 aatccctcgg gcgtaaagtt ccgccacgtc caccgcgaca ccgaccgggc caagtggacc
     1981 acctccgagg cgtccagggg caccgcgaac ttgtggttca cggcgagcgc ataggcgtag
     2041 gtggcctgtt cccaagcatc cggtcccgct tcccagtgct tccgaatcgc ccggtaggac
     2101 cacagccgaa cgtcgttgaa cagggccacg ttgcggccca ggccggagtt gtccgctgca
     2161 cggcggggca ggctgcgggg tagccagccg gaaagctggg tggcgagctc tcccagcgtc
     2221 cacaggtggt cattggtcca gaccgtatcc cagccgtcat ggatggggtt cttggtcatg
     2281 aagcccgcgt aaccgaggtc agcgcgtagc gcattgacca gaccccgctc gacgcgggcc
     2341 gcgaacttca tcggctccag ccgcgcattg tccgtgcggc agacgggagc cgccagcagg
     2401 taggccacgt gggcgtggcc ggagggcgac tgcgccacat agttcggcgt cgggacgccg
     2461 ttcagctcca acgccagccc gagggagtcc tcatggtcga ggtcaatgac gatctgcgtg
     2521 gtcaaggctg cggggttggc ctggatgtac cgacgcttga gagcgtcggc gcgggactgg
     2581 cggacaggga accgcccctt gtcatggtca gtggccatgg gccacttctg ccggtagtgc
     2641 tcagcccacc gctccgccgt catcatcaca ggatgacggt atcggcctac ccctcatccc
     2701 tcaggtagga acacaccctt atcagatatt ggtggtcccc aagggtcccc gcacggtttg
     2761 accaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg
     2821 gtcgaatttg ctttcgaatt tctgccattc atccgcttat tatcacttat tcaggcgtag
     2881 caaccaggcg tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac
     2941 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     3001 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     3061 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     3121 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     3181 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     3241 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     3301 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     3361 cggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     3421 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     3481 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     3541 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     3601 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     3661 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc
     3721 ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt
     3781 atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tactgattta gtgtatgatg
     3841 gtgtttttga ggtgctccag tggcttctgt ttctatcagc tgtccctcct gttcagctac
     3901 tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc ggagtgtata
     3961 ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa
     4021 aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc gcttcctcgc
     4081 tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg
     4141 gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc
     4201 aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa
     4261 tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc
     4321 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     4381 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     4441 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     4501 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     4561 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     4621 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     4681 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     4741 ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg
     4801 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     4861 tcatgtttga cagcttatca tcgata
//
