LOCUS       40924_32150        8059 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ78, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8059)
  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 8059)
  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 8059)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8059)
  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..8059
                     /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      complement(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(6072..6929)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6930..7034)
                     /label=AmpR promoter
     misc_feature    7071..7574
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        7839..8059
                     /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 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc atcacgtgct
     7081 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
     7141 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
     7201 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
     7261 ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac
     7321 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
     7381 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
     7441 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     7501 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     7561 acaggcaagc gatccgtcct aagaaaccat tattatcatg acattaacct ataaaaatag
     7621 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7681 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7741 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7801 agagcagatt gtactgagag tgcaccatac cacagctttt caattcaatt catcattttt
     7861 tttttattct tttttttgat ttcggtttct ttgaaatttt tttgattcgg taatctccga
     7921 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
     7981 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacctg
     8041 caggaaacga agataaatc
//
