LOCUS       sequence-365391-        8101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control plasmid that expresses off-target dsRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8101)
  AUTHORS   Leonard SP, Powell JE, Perutka J, Geng P, Heckmann LC, Horak RD, 
            Davies BW, Ellington AD, Barrick JE, Moran NA
  TITLE     Engineered symbionts activate honey bee immunity and limit 
            pathogens.
  JOURNAL   Science. 2020 Jan 31;367(6477):573-576. doi: 
            10.1126/science.aax9039. Epub 2020 Jan 30.
  PUBMED    32001655
REFERENCE   2  (bases 1 to 8101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.aax9039"; journalName: "Science"; date: 
            "2020-01-31- 31"; volume: "367"; issue: "6477"; pages: "573-576"
COMMENT     Sequence Label: pBTK562 (pDS-GFP)
FEATURES             Location/Qualifiers
     source          1..8101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            2210..2297
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      2639..3033
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     CDS             complement(4742..5533)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(5913..6764)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(6751..7590)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(join(8101,1..971))
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
ORIGIN
        1 tacatgctga aatctggccc gcccctgtcc atgcctcgct ggcggggtgc cggtgcccgt
       61 gccagctcgg cccgcgcaag ctggacgctg ggcagaccca tgaccttgct gacggtgcgc
      121 tcgatgtaat ccgcttcgtg gccgggcttg cgctctgcca gcgctgggct ggcctcggcc
      181 atggccttgc cgatttcctc ggcactgcgg ccccggctgg ccagcttctg cgcggcgata
      241 aagtcgcact tgctgaggtc atcaccgaag cgcttgacca gcccggccat ctcgctgcgg
      301 tactcgtcca gcgccgtgcg ccggtggcgg ctaagctgcc gctcgggcag ttcgaggctg
      361 gccagcctgc gggccttctc ctgctgccgc tgggcctgct cgatctgctg gccagcctgc
      421 tgcaccagcg ccgggccagc ggtggcggtc ttgcccttgg attcacgcag cagcacccac
      481 ggctgataac cggcgcgggt ggtgtgcttg tccttgcggt tggtgaagcc cgccaagcgg
      541 ccatagtggc ggctgtcggc gctggccggg tcggcgtcgt actcgctggc cagcgtccgg
      601 gcaatctgcc cccgaagttc accgcctgcg gcgtcggcca ccttgaccca tgcctgatag
      661 ttcttcgggc tggtttccac taccagggca ggctcccggc cctcggcttt catgtcatcc
      721 aggtcaaact cgctgaggtc gtccaccagc accagaccat gccgctcctg ctcggcgggc
      781 ctgatataca cgtcattgcc ctgggcattc atccgcttga gccatggcgt gttctggagc
      841 acttcggcgg ctgaccattc ccggttcatc atctggccgg tggtggcgtc cctgacgccg
      901 atatcgaagc gctcacagcc catggccttg agctgtcggc ctatggcctg caaagtcctg
      961 tcgttcttca tcgggccacc aagcgcagcc agatcgagcc gtcctcggtt gtcagtggcg
     1021 tcaggtcgag caagagcaac gatgcgatca gcagcaccac cgtaggcatc atggaagcca
     1081 gcatcacggt tagccatagc ttccagtgcc acccccgcga cgcgctccgg gcgctctgcg
     1141 cggcgctgct cacctcggcg gctacctccc gcaactcttt ggccagctcc acccatgccg
     1201 cccctgtctg gcgctgggct ttcagccact ccgccgcctg cgcctcgctg gcctgctggg
     1261 tctggctcat gacctgccgg gcttcgtcgg ccagtgtcgc catgctctgg gccagcggtt
     1321 cgatctgctc cgctaactcg ttgatgcctc tggatttctt cactctgtcg attgcgttca
     1381 tggtctattg cctcccggta ttcctgtaag tcgatgatct gggcgttggc ggtgtcgatg
     1441 ttcagggcca cgtctgcccg gtcggtgcgg atgccccggc cttccatctc caccacgttc
     1501 ggccccaggt gaacaccggg caggcgctcg atgccctgcg cctcaagtgt tctgtggtca
     1561 atgcgggcgt cgtggccagc ccgctctaat gcccggttgg catggtcggc ccatgcctcg
     1621 cgggtctgct caagccatgc cttgggcttg agcgcttcgg tcttctgtgc cccgcccttc
     1681 tccggggtct tgccgttgta ccgcttgaac cactgagcgg cgggccgctc gatgccgtca
     1741 ttgatccgct cggagatcat caggtggcag tgcgggttct cgccgccacc ggcatggatg
     1801 gccagcgtat acggcaggcg ctcggcaccg gtcaggtgct gggcgaactc ggacgccagc
     1861 gccttctgct ggtcgagggt cagctcgacc ggcagggcaa attcgacctc cttgaacagc
     1921 cgcccattgg cgcgttcata caggtcggca gcatcccagt agtcggcggg ccgctcgacg
     1981 aactccggca tgtgcccgga ttcggcgtgc aagacttcat ccatgtcgcg ggcatacttg
     2041 ccttcgcgct ggatgtagtc ggccttggcc ctggccgatt ggccgcccga cctgctgccg
     2101 gttttcgccg taaggtgata aatcgccatg ctgcctcgct gttgcttttg cttttcggct
     2161 ccatgcaatg gccctcggag agcgcaccgc ccgaagggtg gccgttaggc cagtttctcg
     2221 aagagaaacc ggtaagtgcg ccctccccta caaagtaggg tcgggattgc cgccgctgtg
     2281 cctccatgat agcctacgag acagcacatt aacaatgggg tgtcaagatg gttaagggga
     2341 gcaacaaggc ggcggatcgg ctggccaagc tcgaagaaca acgagcgcga atcaatgccg
     2401 aaattcagcg ggtgcgggca agggaacagc agcaagagcg caagaacgaa acaaggcgca
     2461 aggtgctggt gggggccatg attttggcca aggtgaacag cagcgagtgg ccggaggatc
     2521 ggctcatggc ggcaatggat gcgtaccttg aacgcgacca cgaccgcgcc ttgttcggtc
     2581 tgccgccacg ccagaaggat gagccgggct gaatgatcga ccgagacagg ccctgcgggg
     2641 ctgcacacgc gcccccaccc ttcgggtagg gggaaaggcc gctaaagcgg ctaaaagcgc
     2701 tccagcgtat ttctgcgggg tttggtgtgg ggtttagcgg gctttgcccg cctttccccc
     2761 tgccgcgcag cggtggggcg gtgtgtagcc tagcgcagcg aatagaccag ctatccggcc
     2821 tctggccggg catattgggc aagggcagca gcgccccaca agggcgctga taaccgcgcc
     2881 tagtggatta ttcttagata atcatggatg gatttttcca acaccccgcc agcccccgcc
     2941 cctgctgggt ttgcaggttt gggggcgtga cagttattgc aggggttcgt gacagttatt
     3001 gcaggggggc gtgacagtta ttgcaggggt tcgtgacagt tagtacggga gtgacgggca
     3061 ctggctggca atgtctagca acggcaggca tttcggctga gggtaaaaga actttccgct
     3121 aagcgataga ctgtatgtaa acacagtatt gcaaggacgc ggaacatgcc tcatgtggcg
     3181 gccaggacgg ccagccggga tcgggatact ggtcgttacc agagccaccg acccgagcaa
     3241 acccttctct atcagatcgt tgacgagtat tacccggcat tcgctgcgct tatggcagag
     3301 cagggaaagg aattgccggg ctatgtgcaa cgggaatttg aagaatttct ccaatgcggg
     3361 cggctggagc atggctttct acgggttcgc tgcgagtctt gccacgccga gcacctggtc
     3421 gctttcagaa atcaatctaa agtatatatg agtaaacttg gtctgacagg cccctgaatt
     3481 cgcatctaga ctgatgagac gtggtagagc cacaaacagc cggtacaagc aacgatctcc
     3541 aggaccatct gaatcatgcg cggatgacac gaactcacga cggcgatcac agacattaac
     3601 ccacagtaca gacactgcga caacgtggca attcgtcgca atacaacgga cagtcattca
     3661 tctttctgcc cctccaaaag caaaaacccg ccgaagcggg tttttacgta aatcaggtga
     3721 aactgaccga ctttggcagt ttattcttga catgtagtga gggggctggt ataatcacat
     3781 agtactgttt atgagtaaag gagaagagct tttcacagga gttgtcccaa tcctcgtgga
     3841 attagacggt gatgttaatg ggcacaagtt ctctgtcagt ggagagggtg aaggcgacgc
     3901 aacatatggc aagctgaccc ttaaatttat ttgcaccacg ggtaaactac ctgttccatg
     3961 gccaacactg gtcactacgt tcgggtatgg ggttcagtgc tttgcgcgct acccagatca
     4021 catgaaacag cacgactttt tcaagagtgc aatgcccgaa ggctatgtac aggagagaac
     4081 catctttttt aaggatgacg gcaactataa gacacgcgcc gaagtgaagt tcgagggtga
     4141 tacccttgtt aatagaatcg agttaaaggg tattgacttt aaggaagatg gaaatatttt
     4201 aggccacaaa ctggaatata actataactc ccataatgtg tacattatgg ccgacaagca
     4261 aaagaacggt atcaaggtta acttcaagat cagacacaac attgaggatg gaagcgttca
     4321 actagccgac cattaccaac aaaacacccc aattggcgat gggcctgtgc tgttaccaga
     4381 caaccattac ctgtccactc aatctgccct ttcgaaagat cccaacgaaa agcgcgacca
     4441 catggtcctt cttgagtttg tcacggctgc tgggattaca cacggcatgg atgaactata
     4501 caaataaatc caacagtact atgtgattat accagccccc tcactacatg tcaagaataa
     4561 actgccaaag gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg cgggtttttt
     4621 tatgggggga gtttagggaa agagcatttg tcagctgcgt ctcaagcagt tacagagatg
     4681 ttacgaaccc ccaggacatc cgagaatgcg aggcgatgga gggtacaacc cgagcggccg
     4741 cttatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact
     4801 gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa
     4861 gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct
     4921 tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat
     4981 ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg
     5041 cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca
     5101 aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg
     5161 ctggctcgaa gatacccgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc
     5221 gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag
     5281 cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag
     5341 ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt
     5401 gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt
     5461 cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca
     5521 ccgcttccct cataatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt
     5581 tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc
     5641 gaggcataga ctgtacccca aaaaaacagt cataacaagc catgaaaacc gccactgcgc
     5701 cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact
     5761 tgcattacag cttacgaacc gaacaggctt atgtccactg ggttcgtgcc ttcatccgtt
     5821 tccacggtgt gcgtcacccg gcaaccttgg gtagcagcga agtcgaggca tttctgtcct
     5881 ggctggtcat gaccaaaatc ccttaacgtg agtcagcctg ccgccttggg ccgggtgatg
     5941 tcgtacttgc ccgccgcgaa ctcggttacc gtccagccca gcgcgaccag ctccggcaac
     6001 gcctcgcgca cccgctggcg gcgcttgcgc atggtcgaac cactggcctc tgacggccag
     6061 acatagccgc acaaggtatc tatggaagcc ttgccggttt tgccggggtc gatccagcca
     6121 cacagccgct ggtgcagcag gcgggcggtt tcgctgtcca gcgcccgcac ctcgtccatg
     6181 ctgatgcgca catgctggcc gccacccatg acggcctgcg cgatcaaggg gttcagggcc
     6241 acgtacaggc gcccgtccgc ctcgtcgctg gcgtactccg acagcagccg aaacccctgc
     6301 cgcttgcggc cattctgggc gatgatggat accttccaaa ggcgctcgat gcagtcctgt
     6361 atgtgcttga gcgccccacc actatcgacc tctgccccga tttcctttgc cagcgcccga
     6421 tagctacctt tgaccaccat ggcatcagcg gtgacggcct cccacttggg ttccaggaac
     6481 agccggagct gccgtccgcc ttcggtcttg ggttccgggc caagcactag gccattaggc
     6541 ccagccatgg ccaccagccc ttgcaggatg cgcagatcat cagcgcccag cggctccggg
     6601 ccgctgaact cgatccgctt gccgtcgccg tagtcatacg tcacgtccag cttgctgcgc
     6661 ttgcgctcgc cccgcttgag ggcacggaac aggccggggg ccagacagtg cgccgggtcg
     6721 tgccggacgt ggctgaggct gtgcttgttc ttaggcttca ccacggggca cccccttgct
     6781 cttgcgctgc ctctccagca cggcgggctt gagcaccccg ccgtcatgcc gcctgaacca
     6841 ccgatcagcg aacggtgcgc catagttggc cttgctcaca ccgaagcgga cgaagaaccg
     6901 gcgctggtcg tcgtccacac cccattcctc ggcctcggcg ctggtcatgc tcgacaggta
     6961 ggactgccag cggatgttat cgaccagtac cgagctgccc cggctggcct gctgctggtc
     7021 gcctgcgccc atcatggccg cgcccttgct ggcatggtgc aggaacacga tagagcaccc
     7081 ggtatcggcg gcgatggcct ccatgcgacc gatgacctgg gccatggggc cgctggcgtt
     7141 ttcttcctcg atgtggaacc ggcgcagcgt gtccagcacc atcaggcggc ggccctcggc
     7201 ggcgcgcttg aggccgtcga accactccgg ggccatgatg ttgggcaggc tgccgatcag
     7261 cggctggatc agcaggccgt cagccacggc ttgccgttcc tcggcgctga ggtgcgcccc
     7321 aagggcgtgc aggcggtgat gaatggcggt gggcgggtct tcggcgggca ggtagatcac
     7381 cgggccggtg ggcagttcgc ccacctccag cagatccggc ccgcctgcaa tctgtgcggc
     7441 cagttgcagg gccagcatgg atttaccggc accaccgggc gacaccagcg ccccgaccgt
     7501 accggccacc atgttgggca aaacgtagtc cagcggtggc ggcgctgctg cgaacgcctc
     7561 cagaatattg ataggcttat gggtagccat tgattgcctc ctttgcaggc agttggtggt
     7621 taggcgctgg cggggtcact acccccgccc tgcgccgctc tgagttcttc caggcactcg
     7681 cgcagcgcct cgtattcgtc gtcggtcagc cagaacttgc gctgacgcat ccctttggcc
     7741 ttcatgcgct cggcatatcg cgcttggcgt acagcgtcag ggctggccag caggtcgccg
     7801 gtctgcttgt ccttttggtc tttcatatca gtcaccgaga aacttgccgg ggccgaaagg
     7861 cttgtcttcg cggaacaagg acaaggtgca gccgtcaagg ttaaggctgg ccatatcagc
     7921 gactgaaaag cggccagcct cggccttgtt tgacgtataa ccaaagccac cgggcaacca
     7981 atagcccttg tcacttttga tcaggtagac cgaccctgaa gcgctttttt cgtattccat
     8041 aaaaccccct tctgtgcgtg agtactcata gtataacagg cgtgagtacc aacgcaagca
     8101 c
//
