LOCUS       40924_32115        6004 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ56, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6004)
  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 6004)
  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 6004)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6004)
  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..6004
                     /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"
     oriT            complement(893..1001)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     protein_bind    complement(1198..1231)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(1282..1633)
                     /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             1647..2477
                     /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     2641..2657
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2667..2685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2694..2801)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2814..2832)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2853..2869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2877..2893)
                     /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(2901..2931)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2946..2967)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3255..3843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4017..4874)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4875..4979)
                     /label=AmpR promoter
     misc_feature    5016..5519
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        5784..6004
                     /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 cagttattca cgtggagctt atcggccagc
      901 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
      961 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     1021 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     1081 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaagtatac
     1141 cttaaccgcc ggctgacccg gcggggacga ggcaagctaa acagatctct agacctaata
     1201 acttcgtata gcatacatta tacgaagtta tattaagggt tgtcgagccg ctggtgccgc
     1261 tggttggacg ccaagggtga atccgcctcg ataccctgat tactcgcttc ctgcgccctc
     1321 tcaggcggcg ataggggact ggtaaaacgg ggattgccca gacgcctccc ccgccccttc
     1381 aggggcacaa atgcggcccc aacggggcca cgtagtggtg cgttttttgc gtttccaccc
     1441 ttttcttcct tttccctttt aaacctttta ggacgtctac aggccacgta atccgtggcc
     1501 tgtagagttt aaaaagggac ggatttgttg ccattaaggg acggatttgt tgttaagaag
     1561 ggacggattt gttgttgtaa agggacggat ttgttgtatt gtgggacgca gatacagtgt
     1621 ccccttatac acaaggaatg tcgaacgtgg cctcaccccc aatggtttac aaaagcaatg
     1681 ccctggtcga ggccgcgtat cgcctcagtg ttcaggaaca gcggatcgtt ctggcctgta
     1741 ttagccaggt gaagaggagc gagcctgtca ccgatgaagt gatgtattca gtgacggcgg
     1801 aggacatagc gacgatggcg ggtgtcccta tcgaatcttc ctacaaccag ctcaaagaag
     1861 cggccctgcg cctgaaacgg cgggaagtcc ggttaaccca agagcccaat ggcaagggga
     1921 aaagaccgag tgtgatgatt accggctggg tgcaaacaat catctaccgg gagggtgagg
     1981 gccgtgtaga actcaggttc accaaagaca tgctgccgta cctgacggaa ctcaccaaac
     2041 agttcaccaa atacgccttg gctgacgtgg ccaagatgga cagcacccac gcgatcaggc
     2101 tttacgagct gctcatgcaa tgggacagca tcggccagcg cgaaatagaa attgaccagc
     2161 tgcgaaagtg gtttcaactg gaaggccggt atccctcgat caaggacttc aagttgcgag
     2221 tgcttgatcc agccgtgacg cagatcaacg agcacagccc gctacaggtg gagtgggcgc
     2281 agcgaaagac cgggcgcaag gtcacacatc tgttgttcag ttttggaccg aagaagcccg
     2341 ccaaggcggt gggtaaggcc ccagcgaagc gcaaggccgg gaagatttca gatgctgaga
     2401 tcgcgaaaca ggctcgccct ggtgagacat gggaagcggc ccgcgctcga ctaacccaga
     2461 tgccgctgga tctggcctag aggccgtggc caccacggcc cggcctgcct ttcaggctgc
     2521 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2581 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2641 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
     2701 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
     2761 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     2821 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2881 tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt
     2941 gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3001 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3061 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3121 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3181 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3241 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3301 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3361 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3421 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3481 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3541 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3601 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3661 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3721 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3781 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3841 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3901 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3961 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4021 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4081 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4141 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4201 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4261 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4321 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4381 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4441 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4501 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4561 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4621 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4681 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4741 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4801 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4861 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4921 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4981 acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata
     5041 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa
     5101 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata
     5161 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc
     5221 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa
     5281 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt
     5341 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt
     5401 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat
     5461 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc
     5521 gtcctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5581 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5641 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5701 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5761 gagagtgcac cataccacag cttttcaatt caattcatca tttttttttt attctttttt
     5821 ttgatttcgg tttctttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg
     5881 aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg
     5941 aaattgccca gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata
     6001 aatc
//
