LOCUS       40924_37458        5302 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pRMU824Tc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5302)
  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 5302)
  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 5302)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5302)
  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..5302
                     /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"
     promoter        690..719
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             767..1954
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(1977..2243)
                     /codon_start=1
                     /gene="repB"
                     /product="DNA-binding protein RepB"
                     /label=repB
                     /protein_id="AER68072.1"
                     /translation="MSALDPRRRKITAREAAEQVGCTPRHIRSVVAEPRHEFLARAAER
                     QRKAADWKDEGLTYREIAERLDCTPKAAENLVLRGRKARKVTA"
     gene            complement(1977..2243)
                     /gene="repB"
                     /label=repB
     CDS             complement(2243..3079)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /protein_id="AER68073.1"
                     /translation="MTAERWAEHYRQKWPMATDHDKGRFPVRQSRADALKRRYIQANPA
                     ALTTQIVIDLDHEDSLGLALELNGVPTPNYVAQSPSGHAHVAYLLAAPVCRTDNARLEP
                     MKFAARVERGLVNALRADLGYAGFMTKNPIHDGWDTVWTNDHLWTLGELATQLSGWLPR
                     SLPRRAADNSGLGRNVALFNDVRLWSYRAIRKHWEAGPDAWEQATYAYALAVNHKFAVP
                     LDASEVVHLARSVSRWTWRNFTPEGFSKVQTRRSQKAAAVRTAEKIVKAELIRSLV"
     gene            complement(2243..3079)
                     /gene="repA"
                     /label=repA
     CDS             complement(3341..3997)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3998..4100)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(4626..5171)
                     /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 ggctcatgtt tgacagctta tcatccgatt agctttaatg
      721 cggtagttta tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa
      781 tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat
      841 gccggtactg ccgggcctct tgcgggatat cgtccattcc gacagcatcg ccagtcacta
      901 tggcgtgctg ctagcgctat atgcgttgat gcaatttcta tgcgcacccg ttctcggagc
      961 actgtccgac cgctttggcc gccgcccagt cctgctcgct tcgctacttg gagccactat
     1021 cgactacgcg atcatggcga ccacacccgt cctgtggatt ctctacgccg gacgcatcgt
     1081 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     1141 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     1201 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttacatgcac cattccttgc
     1261 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     1321 taagggagag cgccgaccca tgcccttgag agccttcaac ccagtcagct ccttccggtg
     1381 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     1441 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     1501 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     1561 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     1621 ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc
     1681 cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc
     1741 caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag
     1801 cctaacttcg atcattggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac
     1861 atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt
     1921 gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccctag gtggcctcag
     1981 gccgtcacct tgcgggcctt gcggccacgc agaaccaggt tctcagcggc cttgggggtg
     2041 caatccagcc gctccgcaat ctcccggtag gtcaggccct cgtccttcca gtccgcggct
     2101 ttacgctggc gctcagccgc tctcgccagg aattcatgtc gaggctcggc aaccacgctc
     2161 cggatatggc gcggggtgca gccgacctgc tcggctgctt ctcgagcggt gatcttccgg
     2221 cggcggggat ctagtgcgct cattagacga ggctcctaat caactcggct ttcacgatct
     2281 tttcggcggt gcggacggcg gcagccttct ggctccgccg tgtctgcact ttgctgaatc
     2341 cctcgggcgt aaagttccgc cacgtccacc gcgacaccga ccgggccaag tggaccacct
     2401 ccgaggcgtc caggggcacc gcgaacttgt ggttcacggc gagcgcatag gcgtaggtgg
     2461 cctgttccca agcatccggt cccgcttccc agtgcttccg aatcgcccgg taggaccaca
     2521 gccgaacgtc gttgaacagg gccacgttgc ggcccaggcc ggagttgtcc gctgcacggc
     2581 ggggcaggct gcggggtagc cagccggaaa gctgggtggc gagctctccc agcgtccaca
     2641 ggtggtcatt ggtccagacc gtatcccagc cgtcatggat ggggttcttg gtcatgaagc
     2701 ccgcgtaacc gaggtcagcg cgtagcgcat tgaccagacc ccgctcgacg cgggccgcga
     2761 acttcatcgg ctccagccgc gcattgtccg tgcggcagac gggagccgcc agcaggtagg
     2821 ccacgtgggc gtggccggag ggcgactgcg ccacatagtt cggcgtcggg acgccgttca
     2881 gctccaacgc cagcccgagg gagtcctcat ggtcgaggtc aatgacgatc tgcgtggtca
     2941 aggctgcggg gttggcctgg atgtaccgac gcttgagagc gtcggcgcgg gactggcgga
     3001 cagggaaccg ccccttgtca tggtcagtgg ccatgggcca cttctgccgg tagtgctcag
     3061 cccaccgctc cgccgtcatc atcacaggat gacggtatcg gcctacccct catccctcag
     3121 gtaggaacac acccttatca gatattggtg gtccccaagg gtccccgcac ggtttgacca
     3181 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     3241 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac
     3301 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
     3361 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     3421 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     3481 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     3541 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     3601 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     3661 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     3721 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga
     3781 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     3841 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     3901 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     3961 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     4021 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     4081 aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
     4141 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
     4201 attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt
     4261 ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac
     4321 ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg
     4381 cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa
     4441 ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac
     4501 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg
     4561 agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag
     4621 ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag
     4681 tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc
     4741 gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt
     4801 ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg
     4861 tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga
     4921 gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta attgatttag
     4981 aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg
     5041 actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg
     5101 aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa
     5161 aacgatctca agaagatcat cttattaatc agataaaata tttctagatt tcagtgcaat
     5221 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     5281 gtttgacagc ttatcatcga ta
//
