LOCUS       40924_32170        5337 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle/suicide vector pMQ87, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5337)
  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 5337)
  AUTHORS   Shanks RMQ., Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-SEP-2005) Microbiology and Immunology, Dartmouth 
            Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 5337)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5337)
  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 
            (15-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..5337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(352..461)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(882..909)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1001..1087)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1437..1453
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(1457..1513)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1523..1539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1547..1563)
                     /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(1571..1601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1616..1637)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1925..2513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2769..3299)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3488..3516)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(3534..3638)
                     /label=AmpR promoter
     CDS             complement(3738..4538)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4539..4759)
                     /label=URA3 promoter
     misc_feature    4787..5290
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgagctaga gtcgatcttc gccagcaggg cgaggatcgt ggcatcaccg aaccgcgccg
       61 tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc caggcggccc aggtcgccat
      121 tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt ttgtagccct
      181 ggccgacggc cagcaggtag gccgacaggc tcatgccggc cgccgccgcc ttttcctcaa
      241 tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg tgggctgccc ttcctggttg
      301 gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc
      361 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
      421 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
      481 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
      541 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaagcgct
      601 gcttccctgc tgttttgtgg aatatctacc gactggaaac aggcaaatgc aggaaattac
      661 tgaactgagg ggacaggcga gagacgatgc caaagagcta caccgacgag ctggccgagt
      721 gggttgaatc ccgcgcggcc aagaagcgcc ggcgtgatga ggctgcggtt gcgttcctgg
      781 cggtgagggc ggatgtcgat atgcgtaagg agaaaatacc gcatcaggcg catatttgaa
      841 tgtatttaga aaaataaaca aaaagagttt gtagaaacgc aaaaaggcca tccgtcagga
      901 tggccttctg cttaatttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
      961 cgggccgttg cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     1021 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     1081 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg
     1141 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
     1201 ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg
     1261 aaaatcttct ctcatccgcc aaaacagcca agctcgccat tcgccattca ggctgcgcaa
     1321 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     1381 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     1441 aacgacggcc agtgccaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac
     1501 cgagctcgaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1741 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1801 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1861 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1921 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1981 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2101 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2341 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     2401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2461 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2521 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2581 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2641 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga caatcgatgc gaattggccg
     2701 cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg
     2761 aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc
     2821 aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca
     2881 taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc
     2941 tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc
     3001 tcaaacctgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca
     3061 gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga
     3121 tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc
     3181 atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca
     3241 cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc
     3301 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
     3361 ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc
     3421 gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt
     3481 gcattacagt ttacgaaccg aacaggctta tgtcaattcg gttgaatact catactcttc
     3541 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3601 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgggt
     3661 aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact
     3721 tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct
     3781 tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca
     3841 acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc
     3901 aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct
     3961 tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc
     4021 ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac
     4081 aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc
     4141 aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct
     4201 gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat
     4261 tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact
     4321 gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg
     4381 ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca
     4441 cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga
     4501 tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag
     4561 gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta
     4621 catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
     4681 gagattaccg aatcaaaaaa atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa
     4741 aatgatgaat tgaattgaaa agctagctta tcgatgggtc cttttcatca cgtgctataa
     4801 aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa
     4861 aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc
     4921 gccaacaaat actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt
     4981 ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat
     5041 cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa
     5101 gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac
     5161 tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac
     5221 acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag
     5281 gcaagcgatc tctaagaaac cattattatc atgacattaa cctataaaaa aggcctc
//
