pMaEvo vector (V004846)

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LOCUS       40924_29666        7375 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mariner mini-transposon delivery vector pMaEvo, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7375)
  AUTHORS   Krol JE, Rogers LM, Krone SM, Top EM.
  TITLE     Dual reporter system for in situ detection of plasmid transfer under
            aerobic and anaerobic conditions
  JOURNAL   Appl. Environ. Microbiol. 76 (13), 4553-4556 (2010)
  PUBMED    20453134
REFERENCE   2  (bases 1 to 7375)
  AUTHORS   Krol JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2010) Biological Sciences, University of Idaho, 
            P.O. Box 445031, Moscow, ID 83844, USA
REFERENCE   3  (bases 1 to 7375)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7375)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2010"; volume: "76"; issue: "13"; 
            pages: "4553-4556"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-MAR-2010) Biological Sciences, University of Idaho, P.O. Box 
            445031, Moscow, ID 83844, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..402
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      1021..1409
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     repeat_region   1450..1477
                     /label=mariner end
                     /note="mariner end"
     protein_bind    complement(1532..1548)
                     /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(1556..1584)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             complement(2053..2844)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        3228..3246
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3247..3271
                     /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)."
     RBS             3286..3308
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3316..3813
                     /codon_start=1
                     /gene="EcFbFP"
                     /product="Evoglow"
                     /label=EcFbFP
                     /protein_id="ADJ00073.1"
                     /translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
                     FVQMTGYETEEILGKNARFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
                     ELNIDPMEIEDKTYFVGIQNDITKQKEYEKLLEILEIPGDPSTCRHASFMLVNRFVKKF
                     LK"
     gene            3316..3813
                     /gene="EcFbFP"
                     /label=EcFbFP
     promoter        4229..4257
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4446..4976
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     repeat_region   5154..5181
                     /label=mariner end
                     /note="mariner end"
     primer_bind     5313..5329
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5339..5357
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5420..6466)
                     /codon_start=1
                     /gene="tnpA"
                     /product="TnpA"
                     /label=tnpA
                     /note="mariner transposase"
                     /protein_id="ADJ00075.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     gene            complement(5420..6466)
                     /gene="tnpA"
                     /label=tnpA
     oriT            7267..7375
                     /label=oriT
                     /note="incP origin of transfer"