LOCUS       40924_32160        8760 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ80, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8760)
  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 8760)
  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 8760)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8760)
  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..8760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..801
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     oriT            complement(893..1001)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    complement(1198..1231)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(1282..1633)
                     /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             1647..2477
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     terminator      complement(2679..2765)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2995..3708)
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      3716..3721
                     /label=artificial RBS
                     /note="artificial RBS"
                     /regulatory_class="ribosome_binding_site"
     promoter        complement(3766..4050)
                     /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             4077..4952
                     /label=araC
                     /note="L-arabinose regulatory protein"
     rep_origin      complement(5310..5898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6110..6640)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(6829..6857)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(6934..8276)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        8540..8760
                     /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 cagttattca cgtggagctt atcggccagc
      901 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
      961 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     1021 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     1081 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaagtatac
     1141 cttaaccgcc ggctgacccg gcggggacga ggcaagctaa acagatctct agacctaata
     1201 acttcgtata gcatacatta tacgaagtta tattaagggt tgtcgagccg ctggtgccgc
     1261 tggttggacg ccaagggtga atccgcctcg ataccctgat tactcgcttc ctgcgccctc
     1321 tcaggcggcg ataggggact ggtaaaacgg ggattgccca gacgcctccc ccgccccttc
     1381 aggggcacaa atgcggcccc aacggggcca cgtagtggtg cgttttttgc gtttccaccc
     1441 ttttcttcct tttccctttt aaacctttta ggacgtctac aggccacgta atccgtggcc
     1501 tgtagagttt aaaaagggac ggatttgttg ccattaaggg acggatttgt tgttaagaag
     1561 ggacggattt gttgttgtaa agggacggat ttgttgtatt gtgggacgca gatacagtgt
     1621 ccccttatac acaaggaatg tcgaacgtgg cctcaccccc aatggtttac aaaagcaatg
     1681 ccctggtcga ggccgcgtat cgcctcagtg ttcaggaaca gcggatcgtt ctggcctgta
     1741 ttagccaggt gaagaggagc gagcctgtca ccgatgaagt gatgtattca gtgacggcgg
     1801 aggacatagc gacgatggcg ggtgtcccta tcgaatcttc ctacaaccag ctcaaagaag
     1861 cggccctgcg cctgaaacgg cgggaagtcc ggttaaccca agagcccaat ggcaagggga
     1921 aaagaccgag tgtgatgatt accggctggg tgcaaacaat catctaccgg gagggtgagg
     1981 gccgtgtaga actcaggttc accaaagaca tgctgccgta cctgacggaa ctcaccaaac
     2041 agttcaccaa atacgccttg gctgacgtgg ccaagatgga cagcacccac gcgatcaggc
     2101 tttacgagct gctcatgcaa tgggacagca tcggccagcg cgaaatagaa attgaccagc
     2161 tgcgaaagtg gtttcaactg gaaggccggt atccctcgat caaggacttc aagttgcgag
     2221 tgcttgatcc agccgtgacg cagatcaacg agcacagccc gctacaggtg gagtgggcgc
     2281 agcgaaagac cgggcgcaag gtcacacatc tgttgttcag ttttggaccg aagaagcccg
     2341 ccaaggcggt gggtaaggcc ccagcgaagc gcaaggccgg gaagatttca gatgctgaga
     2401 tcgcgaaaca ggctcgccct ggtgagacat gggaagcggc ccgcgctcga ctaacccaga
     2461 tgccgctgga tctggcctag aggccgtggc caccacggcc cggcctgcct ttcaggctgc
     2521 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2581 ggggatgtgc tgcaaggatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
     2641 ggccgttgct tcgcaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
     2701 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
     2761 atttgatgcc tggcagttcc ctactctcgc atggggagac cccacactac catcggcgct
     2821 acggcgtttc acttctgagt tcggcatggg gtcaggtggg accaccgcgc tactgccgcc
     2881 aggcaaattc tgttttatca gaccgcttct gcgttctgat ttaatctgta tcaggctgaa
     2941 aatcttctct catccgccaa aacagccaag cttgcatgcc tgcagactag tctatttgta
     3001 tagttcatcc atgccatgtg taatcccagc agctgttaca aactcaagaa ggaccatgtg
     3061 gtctctcttt tcgttgggat ctttcgaaag ggcagattgt gtggacaggt aatggttgtc
     3121 tggtaaaagg acagggccat cgccaattgg agtattttgt tgataatggt ctgctagttg
     3181 aacgcttcca tcttcaatgt tgtgtctaat tttgaagtta actttgattc cattcttttg
     3241 tttgtctgcc atgatgtata cattgtgtga gttatagttg tattccaatt tgtgtccaag
     3301 aatgtttcca tcttctttaa aatcaatacc ttttaactcg attctattaa caagggtatc
     3361 accttcaaac ttgacttcag cacgtgtctt gtagttcccg tcatctttga aaaatatagt
     3421 tctttcctgt acataacctt cgggcatggc actcttgaaa aagtcatgct gtttcatatg
     3481 atctgggtat ctcgcaaagc attgaacacc ataaccgaaa gtagtgacaa gtgttggcca
     3541 tggaacaggt agttttccag tagtgcaaat aaatttaagg gtaagttttc cgtatgttgc
     3601 atcaccttca ccctctccac tgacagaaaa tttgtgccca ttaacatcac catctaattc
     3661 aacaagaatt gggacaactc cagtgaaaag ttcttctcct ttactcatat gtatatctcc
     3721 ttccccgggt accgagctcg aattcgctag cccaaaaaaa cgggtatgga gaaacagtag
     3781 agagttgcga taaaaagcgt caggtaggat ccgctaatct tatggataaa aatgctatgg
     3841 catagcaaag tgtgacgccg tgcaaataat caatgtggac ttttctgccg tgattataga
     3901 cacttttgtt acgcgttttt gtcatggctt tggtcccgct ttgttacaga atgcttttaa
     3961 taagcggggt taccggtttg gttagcgaga agagccagta aaagacgcag tgacggcaat
     4021 gtctgatgca atatggacaa ttggtttctt ctctgaatgg cgggagtatg aaaagtatgg
     4081 ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt taatgcccat ctggtggcgg
     4141 gtttaacgcc gattgaggcc aacggttatc tcgatttttt tatcgaccga ccgctgggaa
     4201 tgaaaggtta tattctcaat ctcaccattc gcggtcaggg ggtggtgaaa aatcagggac
     4261 gagaatttgt ttgccgaccg ggtgatattt tgctgttccc gccaggagag attcatcact
     4321 acggtcgtca tccggaggct cgcgaatggt atcaccagtg ggtttacttt cgtccgcgcg
     4381 cctactggca tgaatggctt aactggccgt caatatttgc caatacgggg ttctttcgcc
     4441 cggatgaagc gcaccagccg catttcagcg acctgtttgg gcaaatcatt aacgccgggc
     4501 aaggggaagg gcgctattcg gagctgctgg cgataaatct gcttgagcaa ttgttactgc
     4561 ggcgcatgga agcgattaac gagtcgctcc atccaccgat ggataatcgg gtacgcgagg
     4621 cttgtcagta catcagcgat cacctggcag acagcaattt tgatatcgcc agcgtcgcac
     4681 agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt ccgccagcag ttagggatta
     4741 gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc gaagctgctt ttgagcacca
     4801 cccggatgcc tatcgccacc gtcggtcgca atgttggttt tgacgatcaa ctctatttct
     4861 cgcgggtatt taaaaaatgc accggggcca gcccgagcga gttccgtgcc ggttgtgaag
     4921 aaaaagtgaa tgatgtagcc gtcaagttgt cataattggt aacgaatcag acaattgacg
     4981 gcttgacgga gtagcatagg gtttgcagaa tccctgcttc gtccatttga caggcacatt
     5041 atgcatcgat taagctgtca aacatgagcc cagctgcatt aatgaatcgg ccaacgcgcg
     5101 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6001 atatgagtaa acttggtctg acaatcgatg cgaattggcc gcggcgttgt gacaatttac
     6061 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     6121 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     6181 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     6241 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     6301 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacctg ggcagaacgt
     6361 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     6421 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     6481 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     6541 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     6601 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     6661 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     6721 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     6781 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     6841 gaacaggctt atgtcaattc ggttgaatac tcatactctt caaaaataaa caaatagggg
     6901 ttccgcgcac atttccccga aaagtgccac ctgaacgaag catctgtgct tcattttgta
     6961 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     7021 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     7081 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     7141 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     7201 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     7261 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     7321 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     7381 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     7441 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     7501 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     7561 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     7621 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     7681 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     7741 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     7801 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     7861 cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagagaa
     7921 taggaacttc ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca
     7981 acgcgagctg cgcacataca gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg
     8041 tatatatata tacatgagaa gaacggcata gtgcgtgttt atgcttaaat gcgtacttat
     8101 atgcgtctat ttatgtagga tgaaaggtag tctagtacct cctgtgatat tatcccattc
     8161 catgcggggt atcgtatgct tccttcagca ctacccttta gctgttctat atgctgccac
     8221 tcctcaattg gattagtctc atccttcaat gctatcattt cctttgatat tggatcatac
     8281 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     8341 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     8401 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     8461 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     8521 gtgcaccata ccacagcttt tcaattcaat tcatcatttt ttttttattc ttttttttga
     8581 tttcggtttc tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     8641 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     8701 tgcccagtat tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc
//
