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"
ORIGIN
        1 gctaccgccg gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg
       61 aagggcagcc cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa
      121 aaggcggcgg cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac
      181 aaaatcacgg gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat caatggcgac
      241 ctgggccgcc tgggcggcct gctgaaactc tggctcaccg acgacccgcg cacggcgcgg
      301 ttcggtgatg ccacgatcct cgccctgctg gcgaagatcg aagagaagca ggacgagctt
      361 ggcaaggtca tgatgggcgt ggtccgcccg agggcagagc catgactttt ttagccgcta
      421 aaacggccgg ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca tcaagaagag
      481 cgacttcgcg gagctggtga agtacatcac cgacgagcaa ggcaagaccg agcgcctggg
      541 tcacgtgcgc gtcacgaact gcgaggcaaa caccctgccc gctgtcatgg ccgaggtgat
      601 ggcgacccag cacggcaaca cccgttccga ggccgacaag acctatcacc tgctggttag
      661 cttccgcgcg ggagagaagc ccgacgcgga gacgttgcgc gcgattgagg accgcatctg
      721 cgctgggctt ggcttcgccg agcatcagcg cgtcagtgcc gtgcatcacg acaccgacaa
      781 cctgcacatc catatcgcca tcaacaagat tcacccgacc cgaaacacca tccatgagcc
      841 gtatcgggcc taccgcgccc tcgctgacct ctgcgcgacg ctcgaacggg actacgggct
      901 tgagcgtgac aatcacgaaa cgcggcagcg cgtttccgag aaccgcgcga acgacatgga
      961 gcggcacgcg ggcgtggaaa gcctggtcgg ctggatcccc acgatgcgtc cggcgtagag
     1021 gatctgaaga tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt
     1081 actaagctct catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc
     1141 taacgtacta agctctcatg gctaacgtac taagctctca tgtttcacgt actaagcttt
     1201 catgtttgac caataaaatt aatataaatc agcaacttaa atagcttcta aggttttaag
     1261 ttttataaga aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg
     1321 gacaacaagc cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg
     1381 agtgacacag gaacacttaa cggctgacat gggaattgtg ccctgtacaa gaaagcaggg
     1441 tcccgggagt aacaggttgg ctgataagtc cccggtctct agtgtacaag aaagctgggt
     1501 gatcctctag agtcctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacatta
     1561 tacgagccga tgattaattg tcaacagctc atttcagaat atttgccaga accgttatga
     1621 tgtcggcgca aaaaacatta tccagaacgg gagtgcgcct tgagcgacac gaattatgca
     1681 gtgatttacg acctgcacag ccataccaca gcttccgatg gctgcctgac gccagaagca
     1741 ttggtgcacc gtgcagtcga tgataagctg tcaaacatga gaattatccg caccaacgcg
     1801 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     1861 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     1921 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     1981 atgccagcca gccagacgca gacgcgccga gacagaactt aatgaattcc gcgaacccca
     2041 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     2101 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     2161 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     2221 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     2281 atgggtcacg acgagatcct cgccgtcggg catccgcgcc ttgagcctgg cgaacagttc
     2341 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     2401 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     2461 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     2521 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     2581 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     2641 cgatagccgc gctgcctcgt cttggagttc attcagggca ccggacaggt cggtcttgac
     2701 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     2761 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     2821 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct
     2881 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
     2941 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
     3001 aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc
     3061 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     3121 tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg
     3181 cttgcggcag cgtgaagcta gcggaattga tctcgatccc gcgaaattaa tacgactcac
     3241 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa
     3301 gaaggagata tacatatggc gtcgttccag tcgttcggca tcccgggcca gctggaagtc
     3361 atcaagaagg cgctggatca cgtgcgcgtc ggcgtggtca tcaccgatcc cgcgctggaa
     3421 gataacccga tcgtctacgt gaaccagggc ttcgtgcaga tgaccggcta cgagaccgag
     3481 gaaatcctgg gcaagaacgc gcgcttcctc caggggaagc acaccgatcc ggcggaagtg
     3541 gacaacatcc gcaccgcgct gcaaaataaa gaaccggtca ccgtgcagat ccagaactac
     3601 aagaaggacg gcacgatgtt ctggaacgaa ctgaacatcg atccgatgga aatcgaggat
     3661 aagacgtatt tcgtcggcat ccagaacgac atcaccaagc agaaggaata tgaaaagctg
     3721 ctcgaaattc tagagattcc cggggatccg tcgacctgca ggcatgcaag ctttatgctt
     3781 gtaaaccgtt ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt
     3841 aattagtaat ataatctatt aaaggtcatt caaaaggtca tccaccggat cagcttagta
     3901 aagccctcgc tagattttaa tgcggatgtt gcgattactt cgccaactat tgcgataaca
     3961 agaaaaagcc agcctttcat ggtatatctc ccaatttgtg tagggcttat tatgcacgct
     4021 taaaaataat aaaagcagac ttgacctgat agtttggctg tgagcaatta tgtgcttagt
     4081 gcatctaacg cttgagttaa gccgcgccgc gaagcggcgt cggcttgaac gaattgttag
     4141 acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     4201 ctgatctgcg cgcgcgtcaa ttctcgaatt gacataagcc tgttcggttc gtaaactgta
     4261 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     4321 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac agtctatgcc
     4381 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     4441 caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     4501 taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat
     4561 ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc
     4621 aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc
     4681 ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt
     4741 ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc gagcaccgga
     4801 ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg
     4861 cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa
     4921 agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac
     4981 aattcgttca agccgagatc ggcttcccgg ccgacgcggg ccgtcgcggc gcttgccggt
     5041 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     5101 gttcgcccgg cttctgtatg gaacgggatg cgtctaaaag cctgctttct agagaccggg
     5161 gacttatcag ccaacctgtt actcccggga cccagctttc ttgtacaggg caccggtgcg
     5221 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     5281 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     5341 atacgactca ctatagggcg aattggagga ttccggtcta acaaagaaaa acacattttt
     5401 ttgtcaaaat tcgtttttat tattcaacat agttcccttc aagagcgata caacgattat
     5461 aacgaccttc caattttttg ataccatttt ggtagtactc cttcggtttt gcctcaaaat
     5521 aggcctcagt ttcggcgatc acctcttcat tgcagccaaa ttttttccct gcgagcatcc
     5581 ttttgaggtc tgagaacaag aaaaagtcgc tgggggccag atctggagaa tacggtgggt
     5641 ggggaagcaa ttcgaagccc aattcatgaa tttttgccat cgttctcaat gacttgtggc
     5701 acggtgcgtt gtcttggtgg aacaacactt ttttcttctt catgtggggc cgttttgccg
     5761 cgatttcgac cttcaaacgc tccaataacg ccatataata gtcactgttg atggtttttc
     5821 ccttctcaag ataatcgata aaaattattc catgcgcatc ccaaaaaaca gaggccatta
     5881 ctttgccagc ggacttttga gtctttccac gcttcggaga cggttcaccg gtcgctgtcc
     5941 actcagccga ctgtcgattg gactcaggag tgtagtgatg gagccatgtt tcatccattg
     6001 tcacatatcg acggaaaaac tcgggtgtat tacgagttaa cagctgcaaa caccgcttag
     6061 aatcatcaac acgtcgttgt ttttggtcaa atgtgagctc gcgcggcacc catttcgcac
     6121 agagcttccg catatccaaa tattgatgaa tgatatgacc aacacgttcc tttgatatct
     6181 ttaaggcctc tgctatctcg atcaacttca ttttacggtc attcaaaatc attttgtgga
     6241 tttttttgat gttttcgtcg gtaaccacct ctttcgggcg tccactgcgt tcaccgtcct
     6301 ccgtgctcat ttcaccacgc ttgaattttg cataccaatc aattattgtt gatttccctg
     6361 gggcagagtc cggaaactca ttatcaagcc aagtttttgc ttccactgta ttttttccct
     6421 tcagaaaaca gtattttatc aaaacacgaa attccttttt ttccatatgt ttcacaataa
     6481 caaaagttgc ttcacaaaag acgctctatc tcacaaacta attgacttac agacgtcaaa
     6541 ttttgacacg aatcatttga aggttggtac tatataaaaa taatatgcat ttaatactag
     6601 cgacgccatc tatgtgtcaa tccatcacac tggcggccct agtggatcct ttttgtccgg
     6661 tgttgggttg aaggtgaagc cggtcggggc cgcagcgggg gccggctttt cagccttgcc
     6721 cccctgcttc ggccgccgtg gctccggcgt cttgggtgcc ggcgcgggtt ccgcagcctt
     6781 ggcctgcggt gcgggcacat cggcgggctt ggccttgatg tgccgcctgg cgtgcgagcg
     6841 gaacgtctcg taggagaact tgaccttccc cgtttcccgc atgtgctccc aaatggtgac
     6901 gagcgcatag ccggacgcta acgccgcctc gacatccgcc ctcaccgcca ggaacgcaac
     6961 cgcagcctca tcacgccggc gcttcttggc cgcgcgggat tcaacccact cggccagctc
     7021 gtcggtgtag ctctttggca tcgtctctcg cctgtcccct cagttcagta atttcctgca
     7081 tttgcctgtt tccagtcggt agatattcca caaaacagca gggaagcagc gcttttccgc
     7141 tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc
     7201 acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg
     7261 tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt
     7321 cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccg
//
