LOCUS       40924_32125        6705 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ64, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6705)
  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 6705)
  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 6705)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6705)
  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..6705
                     /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"
     primer_bind     2727..2743
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2753..2771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2780..2887)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2900..2918)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2939..2955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2963..2979)
                     /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(2987..3017)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3032..3053)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3341..3929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4141..4671)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(4860..4888)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(4965..6307)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6571..6705
                     /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 aggcgattaa gttgggtaac
     2701 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     2761 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta tcgataagct
     2821 tgatatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt
     2881 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     2941 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg
     3001 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt
     3061 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3121 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3781 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3961 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4021 tcttcaccta gatatgagta aacttggtct gacaatcgat gcgaattggc cgcggcgttg
     4081 tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta
     4141 ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt
     4201 atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc
     4261 aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg
     4321 gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct gctcaaacct
     4381 gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc
     4441 gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct gatgttggga
     4501 gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc gcatggattt
     4561 gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc cacctaactt
     4621 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc
     4681 tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca
     4741 tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     4801 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     4861 gtttacgaac cgaacaggct tatgtcaatt cggttgaata ctcatactct tcaaaaataa
     4921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaacgaa gcatctgtgc
     4981 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     5041 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     5101 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     5161 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     5221 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     5281 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     5341 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     5401 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     5461 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     5521 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     5581 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     5641 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     5701 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     5761 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     5821 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     5881 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     5941 tttctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt
     6001 ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg
     6061 cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa
     6121 tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata
     6181 ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta
     6241 tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata
     6301 ttggatcata ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6361 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6421 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6481 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6541 ttgtactgag agtgcaccat accacagctt ttcaattcaa ttcatcattt tttttttatt
     6601 cttttttttg atttcggttt ctttgaaatt tttttgattc ggtaatctcc gaacagaagg
     6661 aagaacgaag gaaggagcac agacttagat tggtatatat acgca
//
