LOCUS       40924_18816        7427 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pEXGm5B, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7427)
  AUTHORS   Kvitko BH, Bruckbauer S, Prucha J, McMillan I, Breland EJ, Lehman S,
            Mladinich K, Choi KH, Karkhoff-Schweizer R, Schweizer HP.
  TITLE     A simple method for construction of pir+ Enterobacterial hosts for 
            maintenance of R6K replicon plasmids
  JOURNAL   BMC Res Notes 5, 157 (2012)
  PUBMED    22433797
REFERENCE   2  (bases 1 to 7427)
  AUTHORS   Kvitko BH, Goodyear A, Propst KL, Dow SW, Schweizer HP.
  TITLE     Burkholderia pseudomallei Known Siderophores and Hemin Uptake Are 
            Dispensable for Lethal Murine Melioidosis
  JOURNAL   PLoS Negl Trop Dis 6 (6), E1715 (2012)
  PUBMED    22745846
REFERENCE   3  (bases 1 to 7427)
  AUTHORS   Kvitko BH, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-AUG-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   4  (bases 1 to 7427)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7427)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC Res 
            Notes 5, 157 (2012)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "PLoS Negl 
            Trop Dis"; date: "2012"; volume: "6"; issue: "6"; pages: "E1715"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (17-AUG-2012) Microbiology Immunology and Pathology, Colorado State 
            University, IDRC at Foothills Campus, Campus Delivery 0922, Fort 
            Collins, CO 80523-0922, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(358..467)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(892..919)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1011..1097)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1447..1463
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1554..1570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1578..1594)
                     /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(1602..1632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1647..1668)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1956..2544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(2690..2739)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        2780..2808
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2997..3527
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        3626..3654
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3980..5788
                     /codon_start=1
                     /gene="uidA"
                     /gene_synonym="gusA"
                     /product="UidA"
                     /EC_number="3.2.1.31"
                     /label=uidA
                     /note="beta-glucuronidase; creates blue precipitate on
                     X-gluc"
                     /protein_id="AFV53142.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQVHGNISPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            3980..5788
                     /gene="uidA"
                     /gene_synonym="gusA"
                     /label=uidA
     terminator      5845..5890
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(5904..7256)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="GATQAFAKETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVSE
                     FDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQ
                     KVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGK
                     HYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHT
                     LRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKK
                     RTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGS
                     KMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKG
                     NNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK"
ORIGIN
        1 cgatcctcta gctagagtcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc
       61 gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt
      121 cgccattgat gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt
      181 agccctggcc gacggccagc aggtaggccg acaggctcat gccggccgcc gccgcctttt
      241 cctcaatcgc tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc
      301 tggttggctt ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg
      361 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
      421 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
      481 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
      541 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
      601 agcgctgctt ccctgctgtt ttgtggaata tctaccgact ggaaacaggc aaatgcagga
      661 aattactgaa ctgaggggac aggcgagaga cgatgccaaa gagctacacc gacgagctgg
      721 ccgagtgggt tgaatcccgc gcggccaaga agcgccggcg tgatgaggct gcggttgcgt
      781 tcctggcggt gagggcggat gtcgatatgc gtaaggagaa aataccgcat caggcgcatg
      841 catatttgaa tgtatttaga aaaataaaca aaaagagttt gtagaaacgc aaaaaggcca
      901 tccgtcagga tggccttctg cttaatttga tgcctggcag tttatggcgg gcgtcctgcc
      961 cgccaccctc cgggccgttg cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta
     1021 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga
     1081 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact
     1141 accatcggcg ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc
     1201 gctactgccg ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg
     1261 tatcaggctg aaaatcttct ctcatccgcc aaaacagcca agctcgccat tcgccattca
     1321 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     1381 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     1441 gacgttgtaa aacgacggcc agtgccaagc tcattaccct gttatcccta cccgggcggc
     1501 cgcctcgaga attcgggatt accctgttat ccctagagct tggcgtaatc atggtcatag
     1561 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1621 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1681 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1741 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1801 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1861 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1921 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1981 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2041 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2101 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2161 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2221 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2281 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2341 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     2401 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2461 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2521 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2581 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2641 acctagatcc ttttgggtac cgagctcgaa ttagcttcaa aagcgctctg aagttcctat
     2701 actttctaga gagaatagga acttcggaat aggaacttca agatcccctg attccctttg
     2761 tcaacagcaa tggatcgaat tgacataagc ctgttcggtt cgtaaactgt aatgcaagta
     2821 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     2881 agtggcggtt ttcatggctt gttatgactg tttttttgta cagtctatgc ctcgggcatc
     2941 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     3001 tacgcagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaggtggct
     3061 caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg
     3121 ctgctcttga tcttttcggt cgtgagttcg gagacgtagc cacctactcc caacatcagc
     3181 cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct
     3241 tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccagg tttgagcagc
     3301 cgcgtagtga gatctatatc tatgatctcg cagtctccgg cgagcaccgg aggcagggca
     3361 ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga
     3421 tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca
     3481 tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa caattcgttc
     3541 aagccgagat cggcttcccg gccgcggagt tgttcggtaa attgtcacaa cgccgcggcc
     3601 aattcgatag tgaacacgaa cccagttgac ataagcctgt tcggttcgta aactgtaatg
     3661 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     3721 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttgtacagt ctatgcctcg
     3781 ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa
     3841 cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggcagccgtt gtgctggtgc
     3901 tttctgatag ttgttgtggg gtaggcagtc agagctcgat ttgcttgtcg ccataataga
     3961 ttcacaagaa ggattcgaca tgttacgtcc tgtagaaacc ccaacccgtg aaatcaaaaa
     4021 actcgacggc ctgtgggcat tcagtctgga tcgcgaaaac tgtggaattg atcagcgttg
     4081 gtgggaaagc gcgttacaag aaagccgggc aattgctgtg ccaggcagtt ttaacgatca
     4141 gttcgccgat gcagatattc gtaattatgc gggcaacgtc tggtatcagc gcgaagtctt
     4201 tataccgaaa ggttgggcag gccagcgtat cgtgctgcgt ttcgatgcgg tcactcatta
     4261 cggcaaagtg tgggtcaata atcaggaagt gatggagcat cagggcggct atacgccatt
     4321 tgaagccgat gtcacgccgt atgttattgc cgggaaaagt gtacgtatca ccgtttgtgt
     4381 gaacaacgaa ctgaactggc agactatccc gccgggaatg gtgattaccg acgaaaacgg
     4441 caagaaaaag cagtcttact tccatgattt ctttaactat gccgggatcc atcgcagcgt
     4501 aatgctctac accacgccga acacctgggt ggacgatatc accgtggtga cgcatgtcgc
     4561 gcaagactgt aaccacgcgt ctgttgactg gcaggtggtg gccaatggtg atgtcagcgt
     4621 tgaactgcgt gatgcggatc aacaggtggt tgcaactgga caaggcacta gcgggacttt
     4681 gcaagtggtg aatccgcacc tctggcaacc gggtgaaggt tatctctatg aactgtgcgt
     4741 cacagccaaa agccagacag agtgtgatat ctacccgctt cgcgtcggca tccggtcagt
     4801 ggcagtgaag ggcgaacagt tcctgattaa ccacaaaccg ttctacttta ctggctttgg
     4861 tcgtcatgaa gatgcggact tacgtggcaa aggattcgat aacgtgctga tggtgcacga
     4921 ccacgcatta atggactgga ttggggccaa ctcctaccgt acctcgcatt acccttacgc
     4981 tgaagagatg ctcgactggg cagatgaaca tggcatcgtg gtgattgatg aaactgctgc
     5041 tgtcggcttt aacctctctt taggcattgg tttcgaagcg ggcaacaagc cgaaagaact
     5101 gtacagcgaa gaggcagtca acggggaaac tcagcaagcg cacttacagg cgattaaaga
     5161 gctgatagcg cgtgacaaaa accacccaag cgtggtgatg tggagtattg ccaacgaacc
     5221 ggatacccgt ccgcaagtgc acgggaatat ttcgccactg gcggaagcaa cgcgtaaact
     5281 cgacccgacg cgtccgatca cctgcgtcaa tgtaatgttc tgcgacgctc acaccgatac
     5341 catcagcgat ctctttgatg tgctgtgcct gaaccgttat tacggatggt atgtccaaag
     5401 cggcgatttg gaaacggcag agaaggtact ggaaaaagaa cttctggcct ggcaggagaa
     5461 actgcatcag ccgattatca tcaccgaata cggcgtggat acgttagccg ggctgcactc
     5521 aatgtacacc gacatgtgga gtgaagagta tcagtgtgca tggctggata tgtatcaccg
     5581 cgtctttgat cgcgtcagcg ccgtcgtcgg tgaacaggta tggaatttcg ccgattttgc
     5641 gacctcgcaa ggcatattgc gcgttggcgg taacaagaaa gggatcttca ctcgcgaccg
     5701 caaaccgaag tcggcggctt ttctgctgca aaaacgctgg actggcatga acttcggtga
     5761 aaaaccgcag cagggaggca aacaatgaat caacaactct cctggcgcac catcgtcggc
     5821 tacagcctcg gtgacgtctc agattaaaac gaaaggccca gtctttcgac tgagcctttc
     5881 gttttatttg aaagctacta ttatttgtta actgttaatt gtccttgttc aaggatgctg
     5941 tctttgacaa cagatgtttt cttgcctttg atgttcagca ggaagctcgg cgcaaacgtt
     6001 gattgtttgt ctgcgtagaa tcctctgttt gtcatatagc ttgtaatcac gacattgttt
     6061 cctttcgctt gaggtacagc gaagtgtgag taagtaaagg ttacatcgtt aggatcaaga
     6121 tccattttta acacaaggcc agttttgttc agcggcttgt atgggccagt taaagaatta
     6181 gaaacataac caagcatgta aatatcgtta gacgtaatgc cgtcaatcgt catttttgat
     6241 ccgcgggagt cagtgaacag gtaccatttg ccgttcattt taaagacgtt cgcgcgttca
     6301 atttcatctg ttactgtgtt agatgcaatc agcggtttca tcactttttt cagtgtgtaa
     6361 tcatcgttta gctcaatcat accgagagcg ccgtttgcta actcagccgt gcgtttttta
     6421 tcgctttgca gaagtttttg actttcttga cggaagaatg atgtgctttt gccatagtat
     6481 gctttgttaa ataaagattc ttcgccttgg tagccatctt cagttccagt gtttgcttca
     6541 aatactaagt atttgtggcc tttatcttct acgtagtgag gatctctcag cgtatggttg
     6601 tcgcctgagc tgtagttgcc ttcatcgatg aactgctgta cattttgata cgtttttccg
     6661 tcaccgtcaa agattgattt ataatcctct acaccgttga tgttcaaaga gctgtctgat
     6721 gctgatacgt taacttgtgc agttgtcagt gtttgtttgc cgtaatgttt accggagaaa
     6781 tcagtgtaga ataaacggat ttttccgtca gatgtaaatg tggctgaacc tgaccattct
     6841 tgtgtttggt cttttaggat agaatcattt gcatcgaatt tgtcgctgtc tttaaagacg
     6901 cggccagcgt ttttccagct gtcaatagaa gtttcgccga ctttttgata gaacatgtaa
     6961 atcgatgtgt catccgcatt tttaggatct ccggctaatg caaagacgat gtggtagccg
     7021 tgatagtttg cgacagtgcc gtcagcgttt tgtaatggcc agctgtccca aacgtccagg
     7081 ccttttgcag aagagatatt tttaattgtg gacgaatcaa attcagaaac ttgatatttt
     7141 tcattttttt gctgttcagg gatttgcagc atatcatggc gtgtaatatg ggaaatgccg
     7201 tatgtttcct tatatggctt ttggttcgtt tctttcgcaa acgcttgagt tgcgccgtcg
     7261 ctctgcgcgt gcgccgacat gcaagcggct gcggtgagcg cggcggcgct cagggccaaa
     7321 cgtttgtgtc tacgggaaac ggcttgtttg gagtacgtca tcgtttcctc ctgtgcagcg
     7381 agtgctaatt atatggacgg gttttgacaa attcaagaag tgactag
//
