LOCUS       40924_32155       10033 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ79, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10033)
  AUTHORS   Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA.
  TITLE     Saccharomyces cerevisiae-based molecular tool kit for manipulation 
            of genes from gram-negative bacteria
  JOURNAL   Appl. Environ. Microbiol. 72 (7), 5027-5036 (2006)
  PUBMED    16820502
REFERENCE   2  (bases 1 to 10033)
  AUTHORS   Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2006) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 10033)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10033)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2006"; volume: "72"; issue: "7"; pages:
            "5027-5036"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-MAY-2006) Microbiology and Immunology, Dartmouth Medical School,
            Vail Building Room 505, Hanover, NH 03755, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..801
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     regulatory      complement(964..997)
                     /label=TEF1 terminator
                     /note="TEF1 terminator"
                     /regulatory_class="terminator"
     misc_recomb     964..997
                     /label=loxP
                     /note="loxP"
     protein_bind    964..997
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            complement(1060..2416)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     protein_bind    complement(2471..2504)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(2555..2906)
                     /direction=LEFT
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             2920..3750
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     terminator      complement(3952..4038)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4268..4981)
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      complement(4989..4994)
                     /label=artificial RBS
                     /note="artificial RBS"
                     /regulatory_class="ribosome_binding_site"
     promoter        complement(5039..5323)
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             5350..6225
                     /label=araC
                     /note="L-arabinose regulatory protein"
     rep_origin      complement(6583..7171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7383..7913)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(8102..8130)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(8207..9549)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        9818..10033
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
       61 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
      121 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
      181 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
      241 tccgccaagt acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca
      301 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
      361 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
      421 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
      481 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
      541 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
      601 acacccggtg tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg
      661 gatgatgtgg tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag
      721 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
      781 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
      841 tcacaaatta gagcttcaat ttaattatat cagttattac cctatgcggt gtgaaatacc
      901 gcacagatgc gtaaggagaa aatagagtgc accatacctc tcaggtggat ctgatatcac
      961 ctaataactt cgtatagcat acattatacg aagttatatt aagggttctc gagagctcgt
     1021 tttcgacact ggatggcggc gttagtatcg aatcgacagc agtatagcga ccagcattca
     1081 catacgattg acgcatgata ttactttctg cgcacttaac ttcgcatctg ggcagatgat
     1141 gtcgaggcga aaaaaaatat aaatcacgct aacatttgat taaaatagaa caactacaat
     1201 ataaaaaaac tatacaaatg acaagttctt gaaaacaaga atctttttat tgtcagtact
     1261 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1321 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1381 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1441 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1501 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     1561 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1621 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1681 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1741 tattcttcta atacctggaa tgctgttttg ccggggatcg cagtggtgag taaccatgca
     1801 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1861 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1921 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     1981 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2041 ggcctcgaaa cgtgagtctt ttccttaccc atggttgttt atgttcggat gtgatgtgag
     2101 aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat
     2161 cctaaccttt tatatttctc tacaggggcg cggcgtgggg acaattcaac gcgtctgtga
     2221 ggggagcgtt tccctgctcg caggtctgca gcgaggagcc gtaatttttg cttcgcgccg
     2281 tgcggccatc aaaatgtatg gatgcaaatg attatacatg gggatgtatg ggctaaatgt
     2341 acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc aagactgtca aggagggtat
     2401 tctgggcctc catgtcgctg gccgggtgac ccggcgggga cgaggcaagc taaacagatc
     2461 tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttgtcgag
     2521 ccgctggtgc cgctggttgg acgccaaggg tgaatccgcc tcgataccct gattactcgc
     2581 ttcctgcgcc ctctcaggcg gcgatagggg actggtaaaa cggggattgc ccagacgcct
     2641 cccccgcccc ttcaggggca caaatgcggc cccaacgggg ccacgtagtg gtgcgttttt
     2701 tgcgtttcca cccttttctt ccttttccct tttaaacctt ttaggacgtc tacaggccac
     2761 gtaatccgtg gcctgtagag tttaaaaagg gacggatttg ttgccattaa gggacggatt
     2821 tgttgttaag aagggacgga tttgttgttg taaagggacg gatttgttgt attgtgggac
     2881 gcagatacag tgtcccctta tacacaagga atgtcgaacg tggcctcacc cccaatggtt
     2941 tacaaaagca atgccctggt cgaggccgcg tatcgcctca gtgttcagga acagcggatc
     3001 gttctggcct gtattagcca ggtgaagagg agcgagcctg tcaccgatga agtgatgtat
     3061 tcagtgacgg cggaggacat agcgacgatg gcgggtgtcc ctatcgaatc ttcctacaac
     3121 cagctcaaag aagcggccct gcgcctgaaa cggcgggaag tccggttaac ccaagagccc
     3181 aatggcaagg ggaaaagacc gagtgtgatg attaccggct gggtgcaaac aatcatctac
     3241 cgggagggtg agggccgtgt agaactcagg ttcaccaaag acatgctgcc gtacctgacg
     3301 gaactcacca aacagttcac caaatacgcc ttggctgacg tggccaagat ggacagcacc
     3361 cacgcgatca ggctttacga gctgctcatg caatgggaca gcatcggcca gcgcgaaata
     3421 gaaattgacc agctgcgaaa gtggtttcaa ctggaaggcc ggtatccctc gatcaaggac
     3481 ttcaagttgc gagtgcttga tccagccgtg acgcagatca acgagcacag cccgctacag
     3541 gtggagtggg cgcagcgaaa gaccgggcgc aaggtcacac atctgttgtt cagttttgga
     3601 ccgaagaagc ccgccaaggc ggtgggtaag gccccagcga agcgcaaggc cgggaagatt
     3661 tcagatgctg agatcgcgaa acaggctcgc cctggtgaga catgggaagc ggcccgcgct
     3721 cgactaaccc agatgccgct ggatctggcc tagaggccgt ggccaccacg gcccggcctg
     3781 cctttcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3841 cagctggcga aagggggatg tgctgcaagg atgcctggca gtttatggcg ggcgtcctgc
     3901 ccgccaccct ccgggccgtt gcttcgcaac gttcaaatcc gctcccggcg gatttgtcct
     3961 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
     4021 agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga gaccccacac
     4081 taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg
     4141 cgctactgcc gccaggcaaa ttctgtttta tcagaccgct tctgcgttct gatttaatct
     4201 gtatcaggct gaaaatcttc tctcatccgc caaaacagcc aagcttgcat gcctgcagac
     4261 tagtctattt gtatagttca tccatgccat gtgtaatccc agcagctgtt acaaactcaa
     4321 gaaggaccat gtggtctctc ttttcgttgg gatctttcga aagggcagat tgtgtggaca
     4381 ggtaatggtt gtctggtaaa aggacagggc catcgccaat tggagtattt tgttgataat
     4441 ggtctgctag ttgaacgctt ccatcttcaa tgttgtgtct aattttgaag ttaactttga
     4501 ttccattctt ttgtttgtct gccatgatgt atacattgtg tgagttatag ttgtattcca
     4561 atttgtgtcc aagaatgttt ccatcttctt taaaatcaat accttttaac tcgattctat
     4621 taacaagggt atcaccttca aacttgactt cagcacgtgt cttgtagttc ccgtcatctt
     4681 tgaaaaatat agttctttcc tgtacataac cttcgggcat ggcactcttg aaaaagtcat
     4741 gctgtttcat atgatctggg tatctcgcaa agcattgaac accataaccg aaagtagtga
     4801 caagtgttgg ccatggaaca ggtagttttc cagtagtgca aataaattta agggtaagtt
     4861 ttccgtatgt tgcatcacct tcaccctctc cactgacaga aaatttgtgc ccattaacat
     4921 caccatctaa ttcaacaaga attgggacaa ctccagtgaa aagttcttct cctttactca
     4981 tatgtatatc tccttccccg ggtaccgagc tcgaattcgc tagcccaaaa aaacgggtat
     5041 ggagaaacag tagagagttg cgataaaaag cgtcaggtag gatccgctaa tcttatggat
     5101 aaaaatgcta tggcatagca aagtgtgacg ccgtgcaaat aatcaatgtg gacttttctg
     5161 ccgtgattat agacactttt gttacgcgtt tttgtcatgg ctttggtccc gctttgttac
     5221 agaatgcttt taataagcgg ggttaccggt ttggttagcg agaagagcca gtaaaagacg
     5281 cagtgacggc aatgtctgat gcaatatgga caattggttt cttctctgaa tggcgggagt
     5341 atgaaaagta tggctgaagc gcaaaatgat cccctgctgc cgggatactc gtttaatgcc
     5401 catctggtgg cgggtttaac gccgattgag gccaacggtt atctcgattt ttttatcgac
     5461 cgaccgctgg gaatgaaagg ttatattctc aatctcacca ttcgcggtca gggggtggtg
     5521 aaaaatcagg gacgagaatt tgtttgccga ccgggtgata ttttgctgtt cccgccagga
     5581 gagattcatc actacggtcg tcatccggag gctcgcgaat ggtatcacca gtgggtttac
     5641 tttcgtccgc gcgcctactg gcatgaatgg cttaactggc cgtcaatatt tgccaatacg
     5701 gggttctttc gcccggatga agcgcaccag ccgcatttca gcgacctgtt tgggcaaatc
     5761 attaacgccg ggcaagggga agggcgctat tcggagctgc tggcgataaa tctgcttgag
     5821 caattgttac tgcggcgcat ggaagcgatt aacgagtcgc tccatccacc gatggataat
     5881 cgggtacgcg aggcttgtca gtacatcagc gatcacctgg cagacagcaa ttttgatatc
     5941 gccagcgtcg cacagcatgt ttgcttgtcg ccgtcgcgtc tgtcacatct tttccgccag
     6001 cagttaggga ttagcgtctt aagctggcgc gaggaccaac gtatcagcca ggcgaagctg
     6061 cttttgagca ccacccggat gcctatcgcc accgtcggtc gcaatgttgg ttttgacgat
     6121 caactctatt tctcgcgggt atttaaaaaa tgcaccgggg ccagcccgag cgagttccgt
     6181 gccggttgtg aagaaaaagt gaatgatgta gccgtcaagt tgtcataatt ggtaacgaat
     6241 cagacaattg acggcttgac ggagtagcat agggtttgca gaatccctgc ttcgtccatt
     6301 tgacaggcac attatgcatc gattaagctg tcaaacatga gcccagctgc attaatgaat
     6361 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7261 gatcttcacc tagatatgag taaacttggt ctgacaatcg atgcgaattg gccgcggcgt
     7321 tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattgt
     7381 taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt
     7441 gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca
     7501 ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc
     7561 cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac
     7621 ctgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc
     7681 gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg
     7741 gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat
     7801 ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac
     7861 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct
     7921 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     7981 catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
     8041 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
     8101 cagtttacga accgaacagg cttatgtcaa ttcggttgaa tactcatact cttcaaaaat
     8161 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaacg aagcatctgt
     8221 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     8281 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     8341 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     8401 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     8461 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     8521 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     8581 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     8641 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     8701 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     8761 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     8821 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     8881 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     8941 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     9001 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     9061 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     9121 ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     9181 actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc
     9241 ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc
     9301 tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta
     9361 aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga
     9421 tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc
     9481 tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga
     9541 tattggatca tactaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9601 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9661 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9721 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9781 gattgtactg agagtgcacc ataccacagc ttttcaattc aattcatcat ttttttttta
     9841 ttcttttttt tgatttcggt ttctttgaaa tttttttgat tcggtaatct ccgaacagaa
     9901 ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga
     9961 agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa
    10021 acgaagataa atc
//
