LOCUS       Exported                6926 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pMK2030, complete sequence.
ACCESSION   EU825945
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6926
                     /organism="Reporter vector pMK2030"
                     /mol_type="other DNA"
                     /db_xref="taxon:541044"
     rep_origin      16..437
                     /label=R6K origin of replication
                     /note="R6K origin of replication"
     rep_origin      20..408
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    438..773
                     /label=RP4 oriT (origin of transfer)
                     /note="RP4 oriT (origin of transfer)"
     oriT            579..688
                     /note="incP origin of transfer"
     CDS             888..2078
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline resistance protein"
                     /label=tetA
                     /note="TetA"
                     /protein_id="ACE95079.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVLFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            888..2078
                     /gene="tetA"
                     /label=tetA
     misc_feature    complement(2198..2312)
                     /label=FRT site
                     /note="FRT site"
     protein_bind    complement(2229..2276)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    2317..2441
                     /label=attR1
                     /note="attR1"
     protein_bind    2317..2441
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /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..3209
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl transferase"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="ACE95080.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            2550..3209
                     /gene="cat"
                     /label=cat
     CDS             3551..3856
                     /codon_start=1
                     /gene="ccdB"
                     /product="DNA gyrase inhibitor"
                     /label=ccdB
                     /note="CcdB"
                     /protein_id="ACE95081.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            3551..3856
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    3897..4021
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(3897..4021)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4026..4140
                     /label=FRT site
                     /note="FRT site"
     protein_bind    4062..4109
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4211..4927
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescent protein"
                     /label=gfp
                     /note="GFP"
                     /protein_id="ACE95082.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            4211..4927
                     /gene="gfp"
                     /label=gfp
     primer_bind     4966..4982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5020..6831
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /note="GusA"
                     /protein_id="ACE95083.1"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            5020..6831
                     /gene="gusA"
                     /label=gusA
     primer_bind     complement(6897..6913)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"