LOCUS       Exported                4252 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMK2017, complete sequence.
ACCESSION   AY423865
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4252)
  AUTHORS   House BL, Mortimer MW, Kahn ML.
  TITLE     New recombination methods for Sinorhizobium meliloti genetics
  JOURNAL   Appl. Environ. Microbiol. 70 (5), 2806-2815 (2004)
  PUBMED    15128536
REFERENCE   2  (bases 1 to 4252)
  AUTHORS   House BL, Mortimer MW, Kahn ML.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-SEP-2003) School of Molecular Biosciences, Washington 
            State University, Abelson Hall Room 406A, Pullman, WA 99164, USA
REFERENCE   3  (bases 1 to 4252)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4252
                     /organism="Cloning vector pMK2017"
                     /lab_host="Sinorhizobium meliloti"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:259260"
     rep_origin      1..824
                     /label=RP4 oriT/R6K oriV
                     /note="RP4 oriT/R6K oriV"
     oriT            93..202
                     /note="incP origin of transfer"
     rep_origin      373..761
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     794..810
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(844..860)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    898..945
                     /label=FRT site
                     /note="FRT site"
     protein_bind    898..945
                     /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    1002..1127
                     /label=attR1
                     /note="attR1"
     protein_bind    1003..1127
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1152..1182
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1236..1895
                     /codon_start=1
                     /gene="cat"
                     /product="CAT"
                     /function="chloramphenicol resistance"
                     /label=cat
                     /note="chloramphenicol acetyl-transferase"
                     /protein_id="AAR86224.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            1236..1895
                     /gene="cat"
                     /label=cat
     CDS             2237..2542
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /note="DNA gyrase inhibitor"
                     /protein_id="AAR86225.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            2237..2542
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    complement(2583..2707)
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(2583..2707)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    complement(2765..2812)
                     /label=FRT site
                     /note="FRT site"
     protein_bind    complement(2765..2812)
                     /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             complement(2955..4145)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /function="tetracycline resistance"
                     /label=tetR
                     /protein_id="AAR86226.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVLFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(2955..4145)
                     /gene="tetR"
                     /label=tetR