LOCUS       40924_37448        4012 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRMU824, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4012)
  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 4012)
  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 4012)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4012)
  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..4012
                     /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             complement(687..953)
                     /codon_start=1
                     /gene="repB"
                     /product="DNA-binding protein RepB"
                     /label=repB
                     /protein_id="AER68062.1"
                     /translation="MSALDPRRRKITAREAAEQVGCTPRHIRSVVAEPRHEFLARAAER
                     QRKAADWKDEGLTYREIAERLDCTPKAAENLVLRGRKARKVTA"
     gene            complement(687..953)
                     /gene="repB"
                     /label=repB
     CDS             complement(953..1792)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /protein_id="AER68063.1"
                     /translation="MMTAERWAEHYRQKWPMATDHDKGRFPVRQSRADALKRRYIQANP
                     AALTTQIVIDLDHEDSLGLALELNGVPTPNYVAQSPSGHAHVAYLLAAPVCRTDNARLE
                     PMKFAARVERGLVNALRADLGYAGFMTKNPIHDGWDTVWTNDHLWTLGELATQLSGWLP
                     RSLPRRAADNSGLGRNVALFNDVRLWSYRAIRKHWEAGPDAWEQATYAYALAVNHKFAV
                     PLDASEVVHLARSVSRWTWRNFTPEGFSKVQTRRSQKAAAVRTAEKIVKAELIRSLV"
     gene            complement(953..1792)
                     /gene="repA"
                     /label=repA
     CDS             complement(2051..2707)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2708..2810)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(3336..3881)
                     /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 gtggcctcag gccgtcacct tgcgggcctt gcggccacgc
      721 agaaccaggt tctcagcggc cttgggggtg caatccagcc gctccgcaat ctcccggtag
      781 gtcaggccct cgtccttcca gtccgcggct ttacgctggc gctcagccgc tctcgccagg
      841 aattcatgtc gaggctcggc aaccacgctc cggatatggc gcggggtgca gccgacctgc
      901 tcggctgctt ctcgagcggt gatcttccgg cggcggggat ctagtgcgct cattagacga
      961 ggctcctaat caactcggct ttcacgatct tttcggcggt gcggacggcg gcagccttct
     1021 ggctccgccg tgtctgcact ttgctgaatc cctcgggcgt aaagttccgc cacgtccacc
     1081 gcgacaccga ccgggccaag tggaccacct ccgaggcgtc caggggcacc gcgaacttgt
     1141 ggttcacggc gagcgcatag gcgtaggtgg cctgttccca agcatccggt cccgcttccc
     1201 agtgcttccg aatcgcccgg taggaccaca gccgaacgtc gttgaacagg gccacgttgc
     1261 ggcccaggcc ggagttgtcc gctgcacggc ggggcaggct gcggggtagc cagccggaaa
     1321 gctgggtggc gagctctccc agcgtccaca ggtggtcatt ggtccagacc gtatcccagc
     1381 cgtcatggat ggggttcttg gtcatgaagc ccgcgtaacc gaggtcagcg cgtagcgcat
     1441 tgaccagacc ccgctcgacg cgggccgcga acttcatcgg ctccagccgc gcattgtccg
     1501 tgcggcagac gggagccgcc agcaggtagg ccacgtgggc gtggccggag ggcgactgcg
     1561 ccacatagtt cggcgtcggg acgccgttca gctccaacgc cagcccgagg gagtcctcat
     1621 ggtcgaggtc aatgacgatc tgcgtggtca aggctgcggg gttggcctgg atgtaccgac
     1681 gcttgagagc gtcggcgcgg gactggcgga cagggaaccg ccccttgtca tggtcagtgg
     1741 ccatgggcca cttctgccgg tagtgctcag cccaccgctc cgccgtcatc atcacaggat
     1801 gacggtatcg gcctacccct catccctcag gtaggaacac acccttatca gatattggtg
     1861 gtccccaagg gtccccgcac ggtttgacca aaccagcaat agacataagc ggctatttaa
     1921 cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc
     1981 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
     2041 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     2101 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     2161 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     2221 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     2281 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     2341 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     2401 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     2461 ccagctcacc gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa
     2521 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     2581 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     2641 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     2701 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     2761 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
     2821 ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt
     2881 ctgcgaagtg atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg
     2941 ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct
     3001 atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc
     3061 ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
     3121 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
     3181 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     3241 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     3301 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     3361 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     3421 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     3481 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     3541 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     3601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     3661 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     3721 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     3781 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     3841 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     3901 agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     3961 gccccatacg atataagttg taattctcat gtttgacagc ttatcatcga ta
//
