LOCUS       Exported                7738 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Transposon mutagenesis vector pRL692, complete sequence.
ACCESSION   AF424805
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7738)
  AUTHORS   Koksharova OA, Wolk CP.
  TITLE     A novel gene that bears a DnaJ motif influences cyanobacterial cell 
            division
  JOURNAL   J. Bacteriol. 184 (19), 5524-5528 (2002)
  PUBMED    12218043
REFERENCE   2  (bases 1 to 7738)
  AUTHORS   Wolk CP.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-SEP-2001) MSU-DOE Plant Research Laboratory, Michigan 
            State University, Wilson Road, E. Lansing, MI 48824, USA
REFERENCE   3  (bases 1 to 7738)
  AUTHORS   Wolk CP.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-MAR-2012) MSU-DOE Plant Research Laboratory, Michigan 
            State University, Wilson Road, E. Lansing, MI 48824, USA
REFERENCE   4  (bases 1 to 7738)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Mar 1, 2012 this sequence version replaced AF424805.1. 
            Transposon-bearing plasmid pRL692, a derivative of pRL1063a (GenBank
            Accession Number U55385), bearing transposon Tn5-692 containing the 
            erm gene from pE194 (GenBank Accession Number J01755) and, 
            downstream from the promoter of ribulose bisphosphate carboxylase 
            from Synechococcus sp. PCC 6301 (GenBank Accession Number X03220), 
            the aadA region from Tn7 (GenBank Accession Number X03043).  The 
            transposon confers resistance to erythromycin, streptomycin and 
            spectinomycin.  The transposase gene has been altered by 
            substitution of a 208-bp Psp1406I fragment with an equal-sized 
            fragment from pRZ5421 (Zhou, M., A. Bhasin, and W.S. Reznikoff, J. 
            Mol. Biol. 276:913-925, 1998).  The inverted repeats are modeled on 
            those of pRZ5421.  In addition, a SpeI site has been inserted 
            between the promoter of the transposase and the nearby terminal 
            inverted repeat of the transposon.
            Transposon Tn5-692 borne by pRL692 efficiently mutagenizes 
            Synechococcus sp. strain PCC 7942 and Anabaena variabilis strain 
            ATCC 29413.  The user is cautioned that this transposon appears able
            to transpose repeatedly.
FEATURES             Location/Qualifiers
     source          1..7738
                     /organism="Transposon mutagenesis vector pRL692"
                     /mol_type="other DNA"
                     /label=a derivative of pRL1063a
                     /note="a derivative of pRL1063a"
                     /db_xref="taxon:175345"
     mobile_element  1..6870
                     /mobile_element_type="transposon:Tn5-692"
     misc_feature    1..19
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             409..1197
                     /codon_start=1
                     /gene="aadA"
                     /product="3(9)-O-nucleotidyltransferase"
                     /label=aadA
                     /note="streptomycin adenylyltransferase; from Tn7"
                     /protein_id="AAL30370.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            409..1197
                     /gene="aadA"
                     /label=aadA
     rep_origin      2639..3227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3478..4212)
                     /codon_start=1
                     /gene="erm"
                     /product="adenine methylase"
                     /label=erm
                     /note="confers upon Staphylococcus aureus resistance to 
                     macrolide, lincosamide, and streptogramin type B (MLS) 
                     antibiotics mediated by a specific N6-dimethylation of 
                     adenine in 23 S rRNA; confers erythromycin-resistance"
                     /protein_id="AAL30371.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
                     NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     gene            complement(3478..4212)
                     /gene="erm"
                     /label=erm
     promoter        4485..4587
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(5347..6777)
                     /codon_start=1
                     /product="mutant Tn5 transposase"
                     /label=mutant Tn5 transposase
                     /protein_id="AAL30372.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDRLAHNERFVVRSKHPRKDVESGL
                     YLIDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTGEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     misc_feature    6852..6870
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     oriT            6924..7702
                     /note="for conjugal transfer of the transposon; derived 
                     from conjugative plasmid RK2"
     CDS             complement(7053..7424)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7457..7566
                     /note="incP origin of transfer"