LOCUS       40924_32110        7277 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ52, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  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 7277)
  AUTHORS   Shanks RMQ., Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-SEP-2005) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 7277)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7277)
  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 
            (16-SEP-2005) 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..7277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..801
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    964..997
                     /label=loxP
                     /note="loxP"
     protein_bind    964..997
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            complement(1060..2416)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     protein_bind    complement(2471..2504)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(2555..2906)
                     /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             2920..3750
                     /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"
     primer_bind     3914..3930
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3940..3958
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(3967..4074)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4087..4105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4126..4142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4150..4166)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4174..4204)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4219..4240)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4528..5116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5290..6147)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6148..6252)
                     /label=AmpR promoter
     misc_feature    6289..6792
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        7057..7277
                     /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 cagttattac cctatgcggt gtgaaatacc
      901 gcacagatgc gtaaggagaa aatagagtgc accatacctc tcaggtggat ctgatatcac
      961 ctaataactt cgtatagcat acattatacg aagttatatt aagggttctc gagagctcgt
     1021 tttcgacact ggatggcggc gttagtatcg aatcgacagc agtatagcga ccagcattca
     1081 catacgattg acgcatgata ttactttctg cgcacttaac ttcgcatctg ggcagatgat
     1141 gtcgaggcga aaaaaaatat aaatcacgct aacatttgat taaaatagaa caactacaat
     1201 ataaaaaaac tatacaaatg acaagttctt gaaaacaaga atctttttat tgtcagtact
     1261 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1321 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1381 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1441 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1501 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     1561 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1621 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1681 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1741 tattcttcta atacctggaa tgctgttttg ccggggatcg cagtggtgag taaccatgca
     1801 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1861 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1921 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     1981 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2041 ggcctcgaaa cgtgagtctt ttccttaccc atggttgttt atgttcggat gtgatgtgag
     2101 aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat
     2161 cctaaccttt tatatttctc tacaggggcg cggcgtgggg acaattcaac gcgtctgtga
     2221 ggggagcgtt tccctgctcg caggtctgca gcgaggagcc gtaatttttg cttcgcgccg
     2281 tgcggccatc aaaatgtatg gatgcaaatg attatacatg gggatgtatg ggctaaatgt
     2341 acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc aagactgtca aggagggtat
     2401 tctgggcctc catgtcgctg gccgggtgac ccggcgggga cgaggcaagc taaacagatc
     2461 tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttgtcgag
     2521 ccgctggtgc cgctggttgg acgccaaggg tgaatccgcc tcgataccct gattactcgc
     2581 ttcctgcgcc ctctcaggcg gcgatagggg actggtaaaa cggggattgc ccagacgcct
     2641 cccccgcccc ttcaggggca caaatgcggc cccaacgggg ccacgtagtg gtgcgttttt
     2701 tgcgtttcca cccttttctt ccttttccct tttaaacctt ttaggacgtc tacaggccac
     2761 gtaatccgtg gcctgtagag tttaaaaagg gacggatttg ttgccattaa gggacggatt
     2821 tgttgttaag aagggacgga tttgttgttg taaagggacg gatttgttgt attgtgggac
     2881 gcagatacag tgtcccctta tacacaagga atgtcgaacg tggcctcacc cccaatggtt
     2941 tacaaaagca atgccctggt cgaggccgcg tatcgcctca gtgttcagga acagcggatc
     3001 gttctggcct gtattagcca ggtgaagagg agcgagcctg tcaccgatga agtgatgtat
     3061 tcagtgacgg cggaggacat agcgacgatg gcgggtgtcc ctatcgaatc ttcctacaac
     3121 cagctcaaag aagcggccct gcgcctgaaa cggcgggaag tccggttaac ccaagagccc
     3181 aatggcaagg ggaaaagacc gagtgtgatg attaccggct gggtgcaaac aatcatctac
     3241 cgggagggtg agggccgtgt agaactcagg ttcaccaaag acatgctgcc gtacctgacg
     3301 gaactcacca aacagttcac caaatacgcc ttggctgacg tggccaagat ggacagcacc
     3361 cacgcgatca ggctttacga gctgctcatg caatgggaca gcatcggcca gcgcgaaata
     3421 gaaattgacc agctgcgaaa gtggtttcaa ctggaaggcc ggtatccctc gatcaaggac
     3481 ttcaagttgc gagtgcttga tccagccgtg acgcagatca acgagcacag cccgctacag
     3541 gtggagtggg cgcagcgaaa gaccgggcgc aaggtcacac atctgttgtt cagttttgga
     3601 ccgaagaagc ccgccaaggc ggtgggtaag gccccagcga agcgcaaggc cgggaagatt
     3661 tcagatgctg agatcgcgaa acaggctcgc cctggtgaga catgggaagc ggcccgcgct
     3721 cgactaaccc agatgccgct ggatctggcc tagaggccgt ggccaccacg gcccggcctg
     3781 cctttcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3841 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3901 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     3961 gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc
     4021 ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt
     4081 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4141 tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg
     4201 taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc
     4261 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4321 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6241 aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat
     6301 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa
     6361 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga
     6421 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt
     6481 gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt
     6541 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta
     6601 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta
     6661 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa
     6721 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac
     6781 aggcaagcga tccgtcctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6841 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6901 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6961 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     7021 agcagattgt actgagagtg caccatacca cagcttttca attcaattca tcattttttt
     7081 tttattcttt tttttgattt cggtttcttt gaaatttttt tgattcggta atctccgaac
     7141 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     7201 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacctgca
     7261 ggaaacgaag ataaatc
//
