LOCUS       40924_32185        8905 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ95, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8905)
  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 8905)
  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 8905)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8905)
  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..8905
                     /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"
     rep_origin      1198..1728
                     /label=oriV
                     /note="incP origin of replication"
     CDS             2120..3265
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     terminator      complement(3525..3611)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3841..4554)
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      complement(4562..4567)
                     /label=artificial RBS
                     /note="artificial RBS"
                     /regulatory_class="ribosome_binding_site"
     promoter        complement(4612..4896)
                     /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             4923..5798
                     /label=araC
                     /note="L-arabinose regulatory protein"
     rep_origin      complement(6156..6744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6918..7775)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7776..7880)
                     /label=AmpR promoter
     misc_feature    7917..8420
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        8685..8905
                     /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 agacctaggc
     1201 cgccggcgtt gtggatacca cgcggaaaac ttggccctca ctgacagatg aggggcggac
     1261 gttgacactt gaggggccga ctcacccggc gcggcgttga cagatgaggg gcaggctcga
     1321 tttcggccgg cgacgtggag ctggccagcc tcgcaaatcg gcgaaaacgc ctgattttac
     1381 gcgagtttcc cacagatgat gtggacaagc ctggggataa gtgccctgcg gtattgacac
     1441 ttgaggggcg cgactactga cagatgaggg gcgcgatcct tgacacttga ggggcagagt
     1501 gatgacagat gaggggcgca cctattgaca tttgaggggc tgtccacagg cagaaaatcc
     1561 agcatttgca agggtttccg cccgtttttc ggccaccgct aacctgtctt ttaacctgct
     1621 tttaaaccaa tatttataaa ccttgttttt aaccagggct gcgccctggc gcgtgaccgc
     1681 gcacgccgaa ggggggtgcc cccccttctc gaaccctccc ggcccgctaa cgcggcaccc
     1741 catcccccca ggggctgcgc ccctcggccg cgaacgacct caccccaaaa atggcagcca
     1801 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta
     1861 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat
     1921 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
     1981 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
     2041 ggatctgatg gcgcagggga tcaagatcga cggatcgatc cggggaatta attccggggc
     2101 aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg aaggcataca ggcaagaact
     2161 gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc gcaagccgca ccgtcatgcg
     2221 tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta cggccaagat
     2281 cgagcgcgac agcgtgcaac tggctccccc tgccctgccc gcgccatcgg ccgccgtgga
     2341 gcgttcgcgt cgtctcgaac aggaggcggc aggtttggcg aagtcgatga ccatcgacac
     2401 gcgaggaact atgacgacca agaagcgaaa aaccgccggc gaggacctgg caaaacaggt
     2461 cagcgaggcc aagcaggccg cgttgctgaa acacacgaag cagcagatca aggaaatgca
     2521 gctttccttg ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc caaacgacac
     2581 ggcccgctct gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg cgctgcaaaa
     2641 caaggtcatt ttccacgtca acaaggacgt gaagatcacc tacaccggcg tcgagctgcg
     2701 ggccgacgat gacgaactgg tgtggcagca ggtgttggag tacgcgaagc gcacccctat
     2761 cggcgagccg atcaccttca cgttctacga gctttgccag gacctgggct ggtcgatcaa
     2821 tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga cggcgatggg
     2881 cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc gcttccgcgt
     2941 cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg aaatcgtcgt
     3001 gctgtttgct ggcgaccact acacgaaatt catatgggag aagtaccgca agctgtcgcc
     3061 gacggcccga cggatgttcg actatttcag ctcgcaccgg gagccgtacc cgctcaagct
     3121 ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt ggcgcgagca
     3181 ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg gtggaacacg cctgggtcaa
     3241 tgatgacctg gtgcattgca aacgctaggg ccttgtgggg tcagttccgg ctgggggttc
     3301 agcagccacc tgcattaatt aactagaggc cgtggccacc acggcccggc ctgcctttca
     3361 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3421 cgaaaggggg atgtgctgca aggatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     3481 cctccgggcc gttgcttcgc aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     3541 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     3601 cgttttattt gatgcctggc agttccctac tctcgcatgg ggagacccca cactaccatc
     3661 ggcgctacgg cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact
     3721 gccgccaggc aaattctgtt ttatcagacc gcttctgcgt tctgatttaa tctgtatcag
     3781 gctgaaaatc ttctctcatc cgccaaaaca gccaagcttg catgcctgca gactagtcta
     3841 tttgtatagt tcatccatgc catgtgtaat cccagcagct gttacaaact caagaaggac
     3901 catgtggtct ctcttttcgt tgggatcttt cgaaagggca gattgtgtgg acaggtaatg
     3961 gttgtctggt aaaaggacag ggccatcgcc aattggagta ttttgttgat aatggtctgc
     4021 tagttgaacg cttccatctt caatgttgtg tctaattttg aagttaactt tgattccatt
     4081 cttttgtttg tctgccatga tgtatacatt gtgtgagtta tagttgtatt ccaatttgtg
     4141 tccaagaatg tttccatctt ctttaaaatc aatacctttt aactcgattc tattaacaag
     4201 ggtatcacct tcaaacttga cttcagcacg tgtcttgtag ttcccgtcat ctttgaaaaa
     4261 tatagttctt tcctgtacat aaccttcggg catggcactc ttgaaaaagt catgctgttt
     4321 catatgatct gggtatctcg caaagcattg aacaccataa ccgaaagtag tgacaagtgt
     4381 tggccatgga acaggtagtt ttccagtagt gcaaataaat ttaagggtaa gttttccgta
     4441 tgttgcatca ccttcaccct ctccactgac agaaaatttg tgcccattaa catcaccatc
     4501 taattcaaca agaattggga caactccagt gaaaagttct tctcctttac tcatatgtat
     4561 atctccttcc ccgggtaccg agctcgaatt cgctagccca aaaaaacggg tatggagaaa
     4621 cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg gataaaaatg
     4681 ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt ctgccgtgat
     4741 tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt tacagaatgc
     4801 ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag acgcagtgac
     4861 ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg agtatgaaaa
     4921 gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat gcccatctgg
     4981 tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc gaccgaccgc
     5041 tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg gtgaaaaatc
     5101 agggacgaga atttgtttgc cgaccgggtg atattttgct gttcccgcca ggagagattc
     5161 atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt tactttcgtc
     5221 cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat acggggttct
     5281 ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa atcattaacg
     5341 ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt gagcaattgt
     5401 tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat aatcgggtac
     5461 gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat atcgccagcg
     5521 tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc cagcagttag
     5581 ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag ctgcttttga
     5641 gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac gatcaactct
     5701 atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc cgtgccggtt
     5761 gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg aatcagacaa
     5821 ttgacggctt gacggagtag catagggttt gcagaatccc tgcttcgtcc atttgacagg
     5881 cacattatgc atcgattaag ctgtcaaaca tgagcccagc tgcattaatg aatcggccaa
     5941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6901 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7021 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7861 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc cttttcatca
     7921 cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag
     7981 aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc
     8041 tagggggatc gccaacaaat actacctttt atcttgctct tcctgctctc aggtattaat
     8101 gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg tgattttaca
     8161 ttttacttat cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata
     8221 tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc
     8281 attccgtaac tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa
     8341 ataaataaac acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg
     8401 taagttacag gcaagcgatc cgtcctaaga aaccattatt atcatgacat taacctataa
     8461 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     8521 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     8581 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     8641 ggcatcagag cagattgtac tgagagtgca ccataccaca gcttttcaat tcaattcatc
     8701 attttttttt tattcttttt tttgatttcg gtttctttga aatttttttg attcggtaat
     8761 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat
     8821 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa
     8881 aacctgcagg aaacgaagat aaatc
//
