LOCUS       40924_32140        8039 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ72, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8039)
  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 8039)
  AUTHORS   Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-MAY-2006) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8039)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8039)
  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 
            (17-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..8039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..887
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     oriT            complement(979..1087)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    complement(1284..1317)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(1368..1719)
                     /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             1733..2563
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     terminator      complement(2765..2851)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    complement(3054..3110)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(3131..3415)
                     /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             3442..4317
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     rep_origin      complement(4675..5263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5475..6005)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(6194..6222)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(6299..7641)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        7905..8039
                     /label=URA3 promoter
ORIGIN
        1 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
       61 aaacctgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
      121 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
      181 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
      241 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
      301 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
      361 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
      421 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
      481 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
      541 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
      601 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
      661 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
      721 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
      781 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
      841 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
      901 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
      961 tattcacgtg gagcttatcg gccagcctcg cagagcagga ttcccgttga gcaccgccag
     1021 gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat
     1081 cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa
     1141 tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa
     1201 gcagggttat gcagcggaaa gtatacctta accgccggct gacccggcgg ggacgaggca
     1261 agctaaacag atctctagac ctaataactt cgtatagcat acattatacg aagttatatt
     1321 aagggttgtc gagccgctgg tgccgctggt tggacgccaa gggtgaatcc gcctcgatac
     1381 cctgattact cgcttcctgc gccctctcag gcggcgatag gggactggta aaacggggat
     1441 tgcccagacg cctcccccgc cccttcaggg gcacaaatgc ggccccaacg gggccacgta
     1501 gtggtgcgtt ttttgcgttt ccaccctttt cttccttttc ccttttaaac cttttaggac
     1561 gtctacaggc cacgtaatcc gtggcctgta gagtttaaaa agggacggat ttgttgccat
     1621 taagggacgg atttgttgtt aagaagggac ggatttgttg ttgtaaaggg acggatttgt
     1681 tgtattgtgg gacgcagata cagtgtcccc ttatacacaa ggaatgtcga acgtggcctc
     1741 acccccaatg gtttacaaaa gcaatgccct ggtcgaggcc gcgtatcgcc tcagtgttca
     1801 ggaacagcgg atcgttctgg cctgtattag ccaggtgaag aggagcgagc ctgtcaccga
     1861 tgaagtgatg tattcagtga cggcggagga catagcgacg atggcgggtg tccctatcga
     1921 atcttcctac aaccagctca aagaagcggc cctgcgcctg aaacggcggg aagtccggtt
     1981 aacccaagag cccaatggca aggggaaaag accgagtgtg atgattaccg gctgggtgca
     2041 aacaatcatc taccgggagg gtgagggccg tgtagaactc aggttcacca aagacatgct
     2101 gccgtacctg acggaactca ccaaacagtt caccaaatac gccttggctg acgtggccaa
     2161 gatggacagc acccacgcga tcaggcttta cgagctgctc atgcaatggg acagcatcgg
     2221 ccagcgcgaa atagaaattg accagctgcg aaagtggttt caactggaag gccggtatcc
     2281 ctcgatcaag gacttcaagt tgcgagtgct tgatccagcc gtgacgcaga tcaacgagca
     2341 cagcccgcta caggtggagt gggcgcagcg aaagaccggg cgcaaggtca cacatctgtt
     2401 gttcagtttt ggaccgaaga agcccgccaa ggcggtgggt aaggccccag cgaagcgcaa
     2461 ggccgggaag atttcagatg ctgagatcgc gaaacaggct cgccctggtg agacatggga
     2521 agcggcccgc gctcgactaa cccagatgcc gctggatctg gcctagaggc cgtggccacc
     2581 acggcccggc ctgcctttca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     2641 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggatgcctg gcagtttatg
     2701 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc aacgttcaaa tccgctcccg
     2761 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     2821 agtctttcga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcatgg
     2881 ggagacccca cactaccatc ggcgctacgg cgtttcactt ctgagttcgg catggggtca
     2941 ggtgggacca ccgcgctact gccgccaggc aaattctgtt ttatcagacc gcttctgcgt
     3001 tctgatttaa tctgtatcag gctgaaaatc ttctctcatc cgccaaaaca gccaagcttg
     3061 catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaattc gctagcccaa
     3121 aaaaacgggt atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct
     3181 aatcttatgg ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg
     3241 tggacttttc tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc
     3301 ccgctttgtt acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc
     3361 cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttcttctctg
     3421 aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct gccgggatac
     3481 tcgtttaatg cccatctggt ggcgggttta acgccgattg aggccaacgg ttatctcgat
     3541 ttttttatcg accgaccgct gggaatgaaa ggttatattc tcaatctcac cattcgcggt
     3601 cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga tattttgctg
     3661 ttcccgccag gagagattca tcactacggt cgtcatccgg aggctcgcga atggtatcac
     3721 cagtgggttt actttcgtcc gcgcgcctac tggcatgaat ggcttaactg gccgtcaata
     3781 tttgccaata cggggttctt tcgcccggat gaagcgcacc agccgcattt cagcgacctg
     3841 tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct attcggagct gctggcgata
     3901 aatctgcttg agcaattgtt actgcggcgc atggaagcga ttaacgagtc gctccatcca
     3961 ccgatggata atcgggtacg cgaggcttgt cagtacatca gcgatcacct ggcagacagc
     4021 aattttgata tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg tctgtcacat
     4081 cttttccgcc agcagttagg gattagcgtc ttaagctggc gcgaggacca acgtatcagc
     4141 caggcgaagc tgcttttgag caccacccgg atgcctatcg ccaccgtcgg tcgcaatgtt
     4201 ggttttgacg atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg ggccagcccg
     4261 agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa gttgtcataa
     4321 ttggtaacga atcagacaat tgacggcttg acggagtagc atagggtttg cagaatccct
     4381 gcttcgtcca tttgacaggc acattatgca tcgattaagc tgtcaaacat gagcccagct
     4441 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4501 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4561 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4681 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4741 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4801 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4861 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4921 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4981 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5041 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5101 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5161 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5221 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5281 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5341 attatcaaaa aggatcttca cctagatatg agtaaacttg gtctgacaat cgatgcgaat
     5401 tggccgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc gatctcggct
     5461 tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt
     5521 atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag
     5581 atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca
     5641 atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc
     5701 ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg
     5761 aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc
     5821 ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca
     5881 gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt
     5941 gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt
     6001 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
     6061 ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac
     6121 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     6181 ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aattcggttg aatactcata
     6241 ctcttcaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgaa
     6301 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     6361 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     6421 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
     6481 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     6541 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     6601 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     6661 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     6721 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     6781 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     6841 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     6901 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     6961 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     7021 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     7081 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     7141 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     7201 gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc aaaagcgctc
     7261 tgaagttcct atactttcta gagaatagga acttcggaat aggaacttca aagcgtttcc
     7321 gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt
     7381 cgcacctata tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg
     7441 tgtttatgct taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag
     7501 tacctcctgt gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc
     7561 ctttagctgt tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat
     7621 catttccttt gatattggat catactaaga aaccattatt atcatgacat taacctataa
     7681 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     7741 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7801 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7861 ggcatcagag cagattgtac tgagagtgca ccataccaca gcttttcaat tcaattcatc
     7921 attttttttt tattcttttt tttgatttcg gtttctttga aatttttttg attcggtaat
     7981 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgca
//
