LOCUS       40924_30965        6926 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pMK2030, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  AUTHORS   Humann JL, Schroeder BK, Mortimer MW, House BL, Yurgel SN, Maloney 
            SC, Ward KL, Fallquist HM, Ziemkiewicz HT, Kahn ML.
  TITLE     Construction and expression of sugar kinase transcriptional gene 
            fusions by using the Sinorhizobium meliloti ORFeome
  JOURNAL   Appl. Environ. Microbiol. 74 (21), 6756-6765 (2008)
  PUBMED    18791020
REFERENCE   2  (bases 1 to 6926)
  AUTHORS   Mortimer MW, Kahn ML, Humann JL.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JUN-2008) Institute of Biological Chemistry, 
            Washington State University, Pullman, WA 99164-6340, USA
REFERENCE   3  (bases 1 to 6926)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6926)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "21"; 
            pages: "6756-6765"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-JUN-2008) Institute of Biological Chemistry, Washington State 
            University, Pullman, WA 99164-6340, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(20..408)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            579..688
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             888..2075
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVLFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     protein_bind    complement(2229..2276)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    2317..2436
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2466..2496
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2550..3206
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3551..3853
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3897..4021)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    4062..4109
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4211..4924
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     4966..4982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5020..6828
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     primer_bind     complement(6897..6913)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cggtggaggg gaattcccat gtcagccgtt aagtgttcct gtgtcactca aaattgcttt
       61 gagaggctct aagggcttct cagtgcgtta catccctggc ttgttgtcca caaccgttaa
      121 accttaaaag ctttaaaagc cttatatatt cttttttttc ttataaaact taaaacctta
      181 gaggctattt aagttgctga tttatattaa ttttattgtt caaacatgag agcttagtac
      241 gtgaaacatg agagcttagt acgttagcca tgagagctta gtacgttagc catgagggtt
      301 tagttcgtta aacatgagag cttagtacgt taaacatgag agcttagtac gtgaaacatg
      361 agagcttagt acgtactatc aacaggttga actgctgatc ttcagatcct ctacgccgga
      421 cgcatcgtgg ccggatcaat tccgttttcc gctgcataac cctgcttcgg ggtcattata
      481 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
      541 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
      601 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
      661 gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg
      721 atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcacgga attgatcccc
      781 ctcgaattga cgcgtcaatt ctcatgtttg acagcttatc atcgataagc tagctttaat
      841 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca
      901 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta
      961 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact
     1021 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag
     1081 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta
     1141 tcgactacgc gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg
     1201 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     1261 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     1321 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttacatgca ccattccttg
     1381 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
     1441 ataagggaga gcgccgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     1501 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttgtttatc atgcaactcg
     1561 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     1621 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     1681 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     1741 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
     1801 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
     1861 ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca
     1921 gcctaacttc gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca
     1981 catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt
     2041 tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc
     2101 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
     2161 gcaaaccaac ccttgatcaa gctgacgcgt cctcggtacc gagctcgaat tagcttcaaa
     2221 agcgctctga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaga
     2281 tcccctgatt ccctttgtca acagcaatgg atatcaacaa gtttgtacaa aaaagctgaa
     2341 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
     2401 actacataat actgtaaaac acaacatatc cagtcatatt ggcggccgca ttaggcaccc
     2461 caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt
     2521 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     2581 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     2641 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     2701 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     2761 cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     2821 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     2881 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     2941 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
     3001 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     3061 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     3121 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     3181 actgcgatga gtggcagggc ggggcgtaaa gatctggatc cggcttacta aaagccagat
     3241 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
     3301 cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag
     3361 cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca
     3421 accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa
     3481 gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg
     3541 ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     3601 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
     3661 cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     3721 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
     3781 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     3841 gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata
     3901 gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat
     3961 ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg
     4021 ttgatatcca ttgctgttga caaagggaat caggggatct tgaagttcct attccgaagt
     4081 tcctattctc tagaaagtat aggaacttca gagcgctttt gaagctaatt cgagctcggt
     4141 accgggcccc ccctcgaggg ggggcccggt accagatctt taaatctaga tttaagaagg
     4201 agatatacat atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     4261 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
     4321 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
     4381 gccaacactt gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca
     4441 tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     4501 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
     4561 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     4621 tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca
     4681 aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca
     4741 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     4801 caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca
     4861 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     4921 caaatagtcc agacctgcag gaattcctgc agcccggggg atccacagga aacagctatg
     4981 accatgatta cgccaagctt ggctgcaggt cagtccctta tggtccgtcc tgtagaaacc
     5041 ccaacccgtg aaatcaaaaa actcgacggc ctgtgggcat tcagtctgga tcgcgaaaac
     5101 tgtggaattg atcagcgttg gtgggaaagc gcgttacaag aaagccgggc aattgctgtg
     5161 ccaggcagtt ttaacgatca gttcgccgat gcagatattc gtaattatgc gggcaacgtc
     5221 tggtatcagc gcgaagtctt tataccgaaa ggttgggcag gccagcgtat cgtgctgcgt
     5281 ttcgatgcgg tcactcatta cggcaaagtg tgggtcaata atcaggaagt gatggagcat
     5341 cagggcggct atacgccatt tgaagccgat gtcacgccgt atgttattgc cgggaaaagt
     5401 gtacgtatca ccgtttgtgt gaacaacgaa ctgaactggc agactatccc gccgggaatg
     5461 gtgattaccg acgaaaacgg caagaaaaag cagtcttact tccatgattt ctttaactat
     5521 gccggaatcc atcgcagcgt aatgctctac accacgccga acacctgggt ggacgatatc
     5581 accgtggtga cgcatgtcgc gcaagactgt aaccacgcgt ctgttgactg gcaggtggtg
     5641 gccaatggtg atgtcagcgt tgaactgcgt gatgcggatc aacaggtggt tgcaactgga
     5701 caaggcacta gcgggacttt gcaagtggtg aatccgcacc tctggcaacc gggtgaaggt
     5761 tatctctatg aactgtgcgt cacagccaaa agccagacag agtgtgatat ctacccgctt
     5821 cgcgtcggca tccggtcagt ggcagtgaag ggcgaacagt tcctgattaa ccacaaaccg
     5881 ttctacttta ctggctttgg tcgtcatgaa gatgcggact tacgtggcaa aggattcgat
     5941 aacgtgctga tggtgcacga ccacgcatta atggactgga ttggggccaa ctcctaccgt
     6001 acctcgcatt acccttacgc tgaagagatg ctcgactggg cagatgaaca tggcatcgtg
     6061 gtgattgatg aaactgctgc tgtcggcttt aacctctctt taggcattgg tttcgaagcg
     6121 ggcaacaagc cgaaagaact gtacagcgaa gaggcagtca acggggaaac tcagcaagcg
     6181 cacttacagg cgattaaaga gctgatagcg cgtgacaaaa accacccaag cgtggtgatg
     6241 tggagtattg ccaacgaacc ggatacccgt ccgcaaggtg cacgggaata tttcgcgcca
     6301 ctggcggaag caacgcgtaa actcgacccg acgcgtccga tcacctgcgt caatgtaatg
     6361 ttctgcgacg ctcacaccga taccatcagc gatctctttg atgtgctgtg cctgaaccgt
     6421 tattacggat ggtatgtcca aagcggcgat ttggaaacgg cagagaaggt actggaaaaa
     6481 gaacttctgg cctggcagga gaaactgcat cagccgatta tcatcaccga atacggcgtg
     6541 gatacgttag ccgggctgca ctcaatgtac accgacatgt ggagtgaaga gtatcagtgt
     6601 gcatggctgg atatgtatca ccgcgtcttt gatcgcgtca gcgccgtcgt cggtgaacag
     6661 gtatggaatt tcgccgattt tgcgacctcg caaggcatat tgcgcgttgg cggtaacaag
     6721 aaagggatct tcactcgcga ccgcaaaccg aagtcggcgg cttttctgct gcaaaaacgc
     6781 tggactggca tgaacttcgg tgaaaaaccg cagcagggag gcaaacaatg aatcaactct
     6841 cctggcgcac catggtcggc tacagcctcg ggaattgctg caggtcgact ctagaggatc
     6901 cactagttct agagcggccg ccaccg
//
