LOCUS       40924_32135        9312 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ71, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9312)
  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 9312)
  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 9312)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9312)
  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..9312
                     /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"
     protein_bind    1050..1083
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            complement(1146..2502)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     protein_bind    complement(2557..2590)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(2641..2992)
                     /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             3006..3836
                     /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(4038..4124)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    complement(4327..4383)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(4404..4688)
                     /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             4715..5590
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     rep_origin      complement(5948..6536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6748..7278)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(7467..7495)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(7572..8914)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        9178..9312
                     /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 tattacccta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata gagtgcacca
     1021 tacctctcag gtggatctga tatcacctaa taacttcgta tagcatacat tatacgaagt
     1081 tatattaagg gttctcgaga gctcgttttc gacactggat ggcggcgtta gtatcgaatc
     1141 gacagcagta tagcgaccag cattcacata cgattgacgc atgatattac tttctgcgca
     1201 cttaacttcg catctgggca gatgatgtcg aggcgaaaaa aaatataaat cacgctaaca
     1261 tttgattaaa atagaacaac tacaatataa aaaaactata caaatgacaa gttcttgaaa
     1321 acaagaatct ttttattgtc agtactgatt agaaaaactc atcgagcatc aaatgaaact
     1381 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     1441 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     1501 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     1561 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     1621 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     1681 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     1741 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     1801 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttgccgg
     1861 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     1921 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     1981 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     2041 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     2101 cagcatccat gttggaattt aatcgcggcc tcgaaacgtg agtcttttcc ttacccatgg
     2161 ttgtttatgt tcggatgtga tgtgagaact gtatcctagc aagattttaa aaggaagtat
     2221 atgaaagaag aacctcagtg gcaaatccta accttttata tttctctaca ggggcgcggc
     2281 gtggggacaa ttcaacgcgt ctgtgagggg agcgtttccc tgctcgcagg tctgcagcga
     2341 ggagccgtaa tttttgcttc gcgccgtgcg gccatcaaaa tgtatggatg caaatgatta
     2401 tacatgggga tgtatgggct aaatgtacgg gcgacagtca catcatgccc ctgagctgcg
     2461 cacgtcaaga ctgtcaagga gggtattctg ggcctccatg tcgctggccg ggtgacccgg
     2521 cggggacgag gcaagctaaa cagatctcta gacctaataa cttcgtatag catacattat
     2581 acgaagttat attaagggtt gtcgagccgc tggtgccgct ggttggacgc caagggtgaa
     2641 tccgcctcga taccctgatt actcgcttcc tgcgccctct caggcggcga taggggactg
     2701 gtaaaacggg gattgcccag acgcctcccc cgccccttca ggggcacaaa tgcggcccca
     2761 acggggccac gtagtggtgc gttttttgcg tttccaccct tttcttcctt ttccctttta
     2821 aaccttttag gacgtctaca ggccacgtaa tccgtggcct gtagagttta aaaagggacg
     2881 gatttgttgc cattaaggga cggatttgtt gttaagaagg gacggatttg ttgttgtaaa
     2941 gggacggatt tgttgtattg tgggacgcag atacagtgtc cccttataca caaggaatgt
     3001 cgaacgtggc ctcaccccca atggtttaca aaagcaatgc cctggtcgag gccgcgtatc
     3061 gcctcagtgt tcaggaacag cggatcgttc tggcctgtat tagccaggtg aagaggagcg
     3121 agcctgtcac cgatgaagtg atgtattcag tgacggcgga ggacatagcg acgatggcgg
     3181 gtgtccctat cgaatcttcc tacaaccagc tcaaagaagc ggccctgcgc ctgaaacggc
     3241 gggaagtccg gttaacccaa gagcccaatg gcaaggggaa aagaccgagt gtgatgatta
     3301 ccggctgggt gcaaacaatc atctaccggg agggtgaggg ccgtgtagaa ctcaggttca
     3361 ccaaagacat gctgccgtac ctgacggaac tcaccaaaca gttcaccaaa tacgccttgg
     3421 ctgacgtggc caagatggac agcacccacg cgatcaggct ttacgagctg ctcatgcaat
     3481 gggacagcat cggccagcgc gaaatagaaa ttgaccagct gcgaaagtgg tttcaactgg
     3541 aaggccggta tccctcgatc aaggacttca agttgcgagt gcttgatcca gccgtgacgc
     3601 agatcaacga gcacagcccg ctacaggtgg agtgggcgca gcgaaagacc gggcgcaagg
     3661 tcacacatct gttgttcagt tttggaccga agaagcccgc caaggcggtg ggtaaggccc
     3721 cagcgaagcg caaggccggg aagatttcag atgctgagat cgcgaaacag gctcgccctg
     3781 gtgagacatg ggaagcggcc cgcgctcgac taacccagat gccgctggat ctggcctaga
     3841 ggccgtggcc accacggccc ggcctgcctt tcaggctgcg caactgttgg gaagggcgat
     3901 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggatgc
     3961 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc
     4021 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     4081 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     4141 tactctcgca tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt
     4201 cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag
     4261 accgcttctg cgttctgatt taatctgtat caggctgaaa atcttctctc atccgccaaa
     4321 acagccaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa
     4381 ttcgctagcc caaaaaaacg ggtatggaga aacagtagag agttgcgata aaaagcgtca
     4441 ggtaggatcc gctaatctta tggataaaaa tgctatggca tagcaaagtg tgacgccgtg
     4501 caaataatca atgtggactt ttctgccgtg attatagaca cttttgttac gcgtttttgt
     4561 catggctttg gtcccgcttt gttacagaat gcttttaata agcggggtta ccggtttggt
     4621 tagcgagaag agccagtaaa agacgcagtg acggcaatgt ctgatgcaat atggacaatt
     4681 ggtttcttct ctgaatggcg ggagtatgaa aagtatggct gaagcgcaaa atgatcccct
     4741 gctgccggga tactcgttta atgcccatct ggtggcgggt ttaacgccga ttgaggccaa
     4801 cggttatctc gattttttta tcgaccgacc gctgggaatg aaaggttata ttctcaatct
     4861 caccattcgc ggtcaggggg tggtgaaaaa tcagggacga gaatttgttt gccgaccggg
     4921 tgatattttg ctgttcccgc caggagagat tcatcactac ggtcgtcatc cggaggctcg
     4981 cgaatggtat caccagtggg tttactttcg tccgcgcgcc tactggcatg aatggcttaa
     5041 ctggccgtca atatttgcca atacggggtt ctttcgcccg gatgaagcgc accagccgca
     5101 tttcagcgac ctgtttgggc aaatcattaa cgccgggcaa ggggaagggc gctattcgga
     5161 gctgctggcg ataaatctgc ttgagcaatt gttactgcgg cgcatggaag cgattaacga
     5221 gtcgctccat ccaccgatgg ataatcgggt acgcgaggct tgtcagtaca tcagcgatca
     5281 cctggcagac agcaattttg atatcgccag cgtcgcacag catgtttgct tgtcgccgtc
     5341 gcgtctgtca catcttttcc gccagcagtt agggattagc gtcttaagct ggcgcgagga
     5401 ccaacgtatc agccaggcga agctgctttt gagcaccacc cggatgccta tcgccaccgt
     5461 cggtcgcaat gttggttttg acgatcaact ctatttctcg cgggtattta aaaaatgcac
     5521 cggggccagc ccgagcgagt tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt
     5581 caagttgtca taattggtaa cgaatcagac aattgacggc ttgacggagt agcatagggt
     5641 ttgcagaatc cctgcttcgt ccatttgaca ggcacattat gcatcgatta agctgtcaaa
     5701 catgagccca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6361 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6601 ttttggtcat gagattatca aaaaggatct tcacctagat atgagtaaac ttggtctgac
     6661 aatcgatgcg aattggccgc ggcgttgtga caatttaccg aacaactccg cggccgggaa
     6721 gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat
     6781 cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc
     6841 tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat
     6901 gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata
     6961 gatctcacta cgcggctgct caaacctggg cagaacgtaa gccgcgagag cgccaacaac
     7021 cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag
     7081 gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa
     7141 aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat
     7201 gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc
     7261 gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
     7321 gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca
     7381 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
     7441 gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat gtcaattcgg
     7501 ttgaatactc atactcttca aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7561 agtgccacct gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     7621 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     7681 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     7741 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     7801 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     7861 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     7921 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     7981 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     8041 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     8101 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     8161 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     8221 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     8281 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     8341 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     8401 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     8461 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt
     8521 ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact
     8581 tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc
     8641 tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga
     8701 acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg
     8761 aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc
     8821 cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat
     8881 ccttcaatgc tatcatttcc tttgatattg gatcatacta agaaaccatt attatcatga
     8941 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     9001 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     9061 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     9121 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatacc acagcttttc
     9181 aattcaattc atcatttttt ttttattctt ttttttgatt tcggtttctt tgaaattttt
     9241 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     9301 tatatatacg ca
//
