LOCUS       sequence-361458-        7347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control plasmid that does not express dsRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7347)
  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 7347)
  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: pBTK1017 (pNR)
FEATURES             Location/Qualifiers
     source          1..7347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      186..257
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      273..300
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(319..1110)
                     /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(1490..2341)
                     /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(2328..3167)
                     /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(3678..4649)
                     /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"
     oriT            5888..5975
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      6317..6711
                     /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)"
ORIGIN
        1 gctgccaatg agacgacggg gtcatcacgg ctcatcatgc gccaaacaaa tgtgtgcaat
       61 acacgctcgg atgactgcat gatgaccgca ctgactgggg acagcagatc cacctaagcc
      121 tgtgagagaa gcagacaccc gacagatcaa ggcagttaac tagtgcactg cagtacacca
      181 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
      241 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
      301 tttataccga gcggccgctt atttgccgac taccttggtg atctcgcctt tcacgtagtg
      361 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
      421 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
      481 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
      541 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
      601 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
      661 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
      721 atcaatgtcg atcgtggctg gctcgaagat acccgcaaga atgtcattgc gctgccattc
      781 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
      841 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
      901 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
      961 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     1021 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     1081 cgatacttcg gcgatcaccg cttccctcat aatgtttaac tttgttttag ggcgactgcc
     1141 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     1201 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat
     1261 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     1321 tgagcgcata cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt
     1381 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggta gcagcgaagt
     1441 cgaggcattt ctgtcctggc tggtcatgac caaaatccct taacgtgagt cagcctgccg
     1501 ccttgggccg ggtgatgtcg tacttgcccg ccgcgaactc ggttaccgtc cagcccagcg
     1561 cgaccagctc cggcaacgcc tcgcgcaccc gctggcggcg cttgcgcatg gtcgaaccac
     1621 tggcctctga cggccagaca tagccgcaca aggtatctat ggaagccttg ccggttttgc
     1681 cggggtcgat ccagccacac agccgctggt gcagcaggcg ggcggtttcg ctgtccagcg
     1741 cccgcacctc gtccatgctg atgcgcacat gctggccgcc acccatgacg gcctgcgcga
     1801 tcaaggggtt cagggccacg tacaggcgcc cgtccgcctc gtcgctggcg tactccgaca
     1861 gcagccgaaa cccctgccgc ttgcggccat tctgggcgat gatggatacc ttccaaaggc
     1921 gctcgatgca gtcctgtatg tgcttgagcg ccccaccact atcgacctct gccccgattt
     1981 cctttgccag cgcccgatag ctacctttga ccaccatggc atcagcggtg acggcctccc
     2041 acttgggttc caggaacagc cggagctgcc gtccgccttc ggtcttgggt tccgggccaa
     2101 gcactaggcc attaggccca gccatggcca ccagcccttg caggatgcgc agatcatcag
     2161 cgcccagcgg ctccgggccg ctgaactcga tccgcttgcc gtcgccgtag tcatacgtca
     2221 cgtccagctt gctgcgcttg cgctcgcccc gcttgagggc acggaacagg ccgggggcca
     2281 gacagtgcgc cgggtcgtgc cggacgtggc tgaggctgtg cttgttctta ggcttcacca
     2341 cggggcaccc ccttgctctt gcgctgcctc tccagcacgg cgggcttgag caccccgccg
     2401 tcatgccgcc tgaaccaccg atcagcgaac ggtgcgccat agttggcctt gctcacaccg
     2461 aagcggacga agaaccggcg ctggtcgtcg tccacacccc attcctcggc ctcggcgctg
     2521 gtcatgctcg acaggtagga ctgccagcgg atgttatcga ccagtaccga gctgccccgg
     2581 ctggcctgct gctggtcgcc tgcgcccatc atggccgcgc ccttgctggc atggtgcagg
     2641 aacacgatag agcacccggt atcggcggcg atggcctcca tgcgaccgat gacctgggcc
     2701 atggggccgc tggcgttttc ttcctcgatg tggaaccggc gcagcgtgtc cagcaccatc
     2761 aggcggcggc cctcggcggc gcgcttgagg ccgtcgaacc actccggggc catgatgttg
     2821 ggcaggctgc cgatcagcgg ctggatcagc aggccgtcag ccacggcttg ccgttcctcg
     2881 gcgctgaggt gcgccccaag ggcgtgcagg cggtgatgaa tggcggtggg cgggtcttcg
     2941 gcgggcaggt agatcaccgg gccggtgggc agttcgccca cctccagcag atccggcccg
     3001 cctgcaatct gtgcggccag ttgcagggcc agcatggatt taccggcacc accgggcgac
     3061 accagcgccc cgaccgtacc ggccaccatg ttgggcaaaa cgtagtccag cggtggcggc
     3121 gctgctgcga acgcctccag aatattgata ggcttatggg tagccattga ttgcctcctt
     3181 tgcaggcagt tggtggttag gcgctggcgg ggtcactacc cccgccctgc gccgctctga
     3241 gttcttccag gcactcgcgc agcgcctcgt attcgtcgtc ggtcagccag aacttgcgct
     3301 gacgcatccc tttggccttc atgcgctcgg catatcgcgc ttggcgtaca gcgtcagggc
     3361 tggccagcag gtcgccggtc tgcttgtcct tttggtcttt catatcagtc accgagaaac
     3421 ttgccggggc cgaaaggctt gtcttcgcgg aacaaggaca aggtgcagcc gtcaaggtta
     3481 aggctggcca tatcagcgac tgaaaagcgg ccagcctcgg ccttgtttga cgtataacca
     3541 aagccaccgg gcaaccaata gcccttgtca cttttgatca ggtagaccga ccctgaagcg
     3601 cttttttcgt attccataaa acccccttct gtgcgtgagt actcatagta taacaggcgt
     3661 gagtaccaac gcaagcacta catgctgaaa tctggcccgc ccctgtccat gcctcgctgg
     3721 cggggtgccg gtgcccgtgc cagctcggcc cgcgcaagct ggacgctggg cagacccatg
     3781 accttgctga cggtgcgctc gatgtaatcc gcttcgtggc cgggcttgcg ctctgccagc
     3841 gctgggctgg cctcggccat ggccttgccg atttcctcgg cactgcggcc ccggctggcc
     3901 agcttctgcg cggcgataaa gtcgcacttg ctgaggtcat caccgaagcg cttgaccagc
     3961 ccggccatct cgctgcggta ctcgtccagc gccgtgcgcc ggtggcggct aagctgccgc
     4021 tcgggcagtt cgaggctggc cagcctgcgg gccttctcct gctgccgctg ggcctgctcg
     4081 atctgctggc cagcctgctg caccagcgcc gggccagcgg tggcggtctt gcccttggat
     4141 tcacgcagca gcacccacgg ctgataaccg gcgcgggtgg tgtgcttgtc cttgcggttg
     4201 gtgaagcccg ccaagcggcc atagtggcgg ctgtcggcgc tggccgggtc ggcgtcgtac
     4261 tcgctggcca gcgtccgggc aatctgcccc cgaagttcac cgcctgcggc gtcggccacc
     4321 ttgacccatg cctgatagtt cttcgggctg gtttccacta ccagggcagg ctcccggccc
     4381 tcggctttca tgtcatccag gtcaaactcg ctgaggtcgt ccaccagcac cagaccatgc
     4441 cgctcctgct cggcgggcct gatatacacg tcattgccct gggcattcat ccgcttgagc
     4501 catggcgtgt tctggagcac ttcggcggct gaccattccc ggttcatcat ctggccggtg
     4561 gtggcgtccc tgacgccgat atcgaagcgc tcacagccca tggccttgag ctgtcggcct
     4621 atggcctgca aagtcctgtc gttcttcatc gggccaccaa gcgcagccag atcgagccgt
     4681 cctcggttgt cagtggcgtc aggtcgagca agagcaacga tgcgatcagc agcaccaccg
     4741 taggcatcat ggaagccagc atcacggtta gccatagctt ccagtgccac ccccgcgacg
     4801 cgctccgggc gctctgcgcg gcgctgctca cctcggcggc tacctcccgc aactctttgg
     4861 ccagctccac ccatgccgcc cctgtctggc gctgggcttt cagccactcc gccgcctgcg
     4921 cctcgctggc ctgctgggtc tggctcatga cctgccgggc ttcgtcggcc agtgtcgcca
     4981 tgctctgggc cagcggttcg atctgctccg ctaactcgtt gatgcctctg gatttcttca
     5041 ctctgtcgat tgcgttcatg gtctattgcc tcccggtatt cctgtaagtc gatgatctgg
     5101 gcgttggcgg tgtcgatgtt cagggccacg tctgcccggt cggtgcggat gccccggcct
     5161 tccatctcca ccacgttcgg ccccaggtga acaccgggca ggcgctcgat gccctgcgcc
     5221 tcaagtgttc tgtggtcaat gcgggcgtcg tggccagccc gctctaatgc ccggttggca
     5281 tggtcggccc atgcctcgcg ggtctgctca agccatgcct tgggcttgag cgcttcggtc
     5341 ttctgtgccc cgcccttctc cggggtcttg ccgttgtacc gcttgaacca ctgagcggcg
     5401 ggccgctcga tgccgtcatt gatccgctcg gagatcatca ggtggcagtg cgggttctcg
     5461 ccgccaccgg catggatggc cagcgtatac ggcaggcgct cggcaccggt caggtgctgg
     5521 gcgaactcgg acgccagcgc cttctgctgg tcgagggtca gctcgaccgg cagggcaaat
     5581 tcgacctcct tgaacagccg cccattggcg cgttcataca ggtcggcagc atcccagtag
     5641 tcggcgggcc gctcgacgaa ctccggcatg tgcccggatt cggcgtgcaa gacttcatcc
     5701 atgtcgcggg catacttgcc ttcgcgctgg atgtagtcgg ccttggccct ggccgattgg
     5761 ccgcccgacc tgctgccggt tttcgccgta aggtgataaa tcgccatgct gcctcgctgt
     5821 tgcttttgct tttcggctcc atgcaatggc cctcggagag cgcaccgccc gaagggtggc
     5881 cgttaggcca gtttctcgaa gagaaaccgg taagtgcgcc ctcccctaca aagtagggtc
     5941 gggattgccg ccgctgtgcc tccatgatag cctacgagac agcacattaa caatggggtg
     6001 tcaagatggt taaggggagc aacaaggcgg cggatcggct ggccaagctc gaagaacaac
     6061 gagcgcgaat caatgccgaa attcagcggg tgcgggcaag ggaacagcag caagagcgca
     6121 agaacgaaac aaggcgcaag gtgctggtgg gggccatgat tttggccaag gtgaacagca
     6181 gcgagtggcc ggaggatcgg ctcatggcgg caatggatgc gtaccttgaa cgcgaccacg
     6241 accgcgcctt gttcggtctg ccgccacgcc agaaggatga gccgggctga atgatcgacc
     6301 gagacaggcc ctgcggggct gcacacgcgc ccccaccctt cgggtagggg gaaaggccgc
     6361 taaagcggct aaaagcgctc cagcgtattt ctgcggggtt tggtgtgggg tttagcgggc
     6421 tttgcccgcc tttccccctg ccgcgcagcg gtggggcggt gtgtagccta gcgcagcgaa
     6481 tagaccagct atccggcctc tggccgggca tattgggcaa gggcagcagc gccccacaag
     6541 ggcgctgata accgcgccta gtggattatt cttagataat catggatgga tttttccaac
     6601 accccgccag cccccgcccc tgctgggttt gcaggtttgg gggcgtgaca gttattgcag
     6661 gggttcgtga cagttattgc aggggggcgt gacagttatt gcaggggttc gtgacagtta
     6721 gtacgggagt gacgggcact ggctggcaat gtctagcaac ggcaggcatt tcggctgagg
     6781 gtaaaagaac tttccgctaa gcgatagact gtatgtaaac acagtattgc aaggacgcgg
     6841 aacatgcctc atgtggcggc caggacggcc agccgggatc gggatactgg tcgttaccag
     6901 agccaccgac ccgagcaaac ccttctctat cagatcgttg acgagtatta cccggcattc
     6961 gctgcgctta tggcagagca gggaaaggaa ttgccgggct atgtgcaacg ggaatttgaa
     7021 gaatttctcc aatgcgggcg gctggagcat ggctttctac gggttcgctg cgagtcttgc
     7081 cacgccgagc acctggtcgc tttcagaaat caatctaaag tatatatgag taaacttggt
     7141 ctgacaggcc cctgaattcg catctagatg gtagagccac aaacagccgg tacaagcaac
     7201 gatctccagg accatctgaa tcatgcgcgg atgacacgaa ctcacgacgg cgatcacaga
     7261 cattaaccca cagtacagac actgcgacaa cgtggcaatt cgtcgcaata ccgtctcact
     7321 gaactggccg ataattgcag acgaacg
//
