LOCUS       sequence-361453-        7822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses double-stranded RNA targeting the Def1 gene from Apis 
            mellifera.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7822)
  AUTHORS   Lariviere PJ, Leonard SP, Horak RD, Powell JE, Barrick JE
  TITLE     Honey bee functional genomics using symbiont-mediated RNAi.
  JOURNAL   Nat Protoc. 2023 Mar;18(3):902-928. doi: 10.1038/s41596-022-00778-4.
            Epub 2022 Dec 2.
  PUBMED    36460809
REFERENCE   2  (bases 1 to 7822)
  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: 
            "10.1038/s41596-022-00778-4"; journalName: "Nat Protoc"; date: 
            "2023-03"; volume: "18"; issue: "3"; pages: "902-928"
COMMENT     Sequence Label: pBTK1024 (pDS-Def1)
FEATURES             Location/Qualifiers
     source          1..7822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            2091..2178
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      2520..2914
                     /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)"
     terminator      4211..4282
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4298..4325
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4344..5135)
                     /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(5515..6366)
                     /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(6353..7192)
                     /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(7703..7822,1..852))
                     /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 ctcgatgtaa tccgcttcgt ggccgggctt gcgctctgcc agcgctgggc tggcctcggc
       61 catggccttg ccgatttcct cggcactgcg gccccggctg gccagcttct gcgcggcgat
      121 aaagtcgcac ttgctgaggt catcaccgaa gcgcttgacc agcccggcca tctcgctgcg
      181 gtactcgtcc agcgccgtgc gccggtggcg gctaagctgc cgctcgggca gttcgaggct
      241 ggccagcctg cgggccttct cctgctgccg ctgggcctgc tcgatctgct ggccagcctg
      301 ctgcaccagc gccgggccag cggtggcggt cttgcccttg gattcacgca gcagcaccca
      361 cggctgataa ccggcgcggg tggtgtgctt gtccttgcgg ttggtgaagc ccgccaagcg
      421 gccatagtgg cggctgtcgg cgctggccgg gtcggcgtcg tactcgctgg ccagcgtccg
      481 ggcaatctgc ccccgaagtt caccgcctgc ggcgtcggcc accttgaccc atgcctgata
      541 gttcttcggg ctggtttcca ctaccagggc aggctcccgg ccctcggctt tcatgtcatc
      601 caggtcaaac tcgctgaggt cgtccaccag caccagacca tgccgctcct gctcggcggg
      661 cctgatatac acgtcattgc cctgggcatt catccgcttg agccatggcg tgttctggag
      721 cacttcggcg gctgaccatt cccggttcat catctggccg gtggtggcgt ccctgacgcc
      781 gatatcgaag cgctcacagc ccatggcctt gagctgtcgg cctatggcct gcaaagtcct
      841 gtcgttcttc atcgggccac caagcgcagc cagatcgagc cgtcctcggt tgtcagtggc
      901 gtcaggtcga gcaagagcaa cgatgcgatc agcagcacca ccgtaggcat catggaagcc
      961 agcatcacgg ttagccatag cttccagtgc cacccccgcg acgcgctccg ggcgctctgc
     1021 gcggcgctgc tcacctcggc ggctacctcc cgcaactctt tggccagctc cacccatgcc
     1081 gcccctgtct ggcgctgggc tttcagccac tccgccgcct gcgcctcgct ggcctgctgg
     1141 gtctggctca tgacctgccg ggcttcgtcg gccagtgtcg ccatgctctg ggccagcggt
     1201 tcgatctgct ccgctaactc gttgatgcct ctggatttct tcactctgtc gattgcgttc
     1261 atggtctatt gcctcccggt attcctgtaa gtcgatgatc tgggcgttgg cggtgtcgat
     1321 gttcagggcc acgtctgccc ggtcggtgcg gatgccccgg ccttccatct ccaccacgtt
     1381 cggccccagg tgaacaccgg gcaggcgctc gatgccctgc gcctcaagtg ttctgtggtc
     1441 aatgcgggcg tcgtggccag cccgctctaa tgcccggttg gcatggtcgg cccatgcctc
     1501 gcgggtctgc tcaagccatg ccttgggctt gagcgcttcg gtcttctgtg ccccgccctt
     1561 ctccggggtc ttgccgttgt accgcttgaa ccactgagcg gcgggccgct cgatgccgtc
     1621 attgatccgc tcggagatca tcaggtggca gtgcgggttc tcgccgccac cggcatggat
     1681 ggccagcgta tacggcaggc gctcggcacc ggtcaggtgc tgggcgaact cggacgccag
     1741 cgccttctgc tggtcgaggg tcagctcgac cggcagggca aattcgacct ccttgaacag
     1801 ccgcccattg gcgcgttcat acaggtcggc agcatcccag tagtcggcgg gccgctcgac
     1861 gaactccggc atgtgcccgg attcggcgtg caagacttca tccatgtcgc gggcatactt
     1921 gccttcgcgc tggatgtagt cggccttggc cctggccgat tggccgcccg acctgctgcc
     1981 ggttttcgcc gtaaggtgat aaatcgccat gctgcctcgc tgttgctttt gcttttcggc
     2041 tccatgcaat ggccctcgga gagcgcaccg cccgaagggt ggccgttagg ccagtttctc
     2101 gaagagaaac cggtaagtgc gccctcccct acaaagtagg gtcgggattg ccgccgctgt
     2161 gcctccatga tagcctacga gacagcacat taacaatggg gtgtcaagat ggttaagggg
     2221 agcaacaagg cggcggatcg gctggccaag ctcgaagaac aacgagcgcg aatcaatgcc
     2281 gaaattcagc gggtgcgggc aagggaacag cagcaagagc gcaagaacga aacaaggcgc
     2341 aaggtgctgg tgggggccat gattttggcc aaggtgaaca gcagcgagtg gccggaggat
     2401 cggctcatgg cggcaatgga tgcgtacctt gaacgcgacc acgaccgcgc cttgttcggt
     2461 ctgccgccac gccagaagga tgagccgggc tgaatgatcg accgagacag gccctgcggg
     2521 gctgcacacg cgcccccacc cttcgggtag ggggaaaggc cgctaaagcg gctaaaagcg
     2581 ctccagcgta tttctgcggg gtttggtgtg gggtttagcg ggctttgccc gcctttcccc
     2641 ctgccgcgca gcggtggggc ggtgtgtagc ctagcgcagc gaatagacca gctatccggc
     2701 ctctggccgg gcatattggg caagggcagc agcgccccac aagggcgctg ataaccgcgc
     2761 ctagtggatt attcttagat aatcatggat ggatttttcc aacaccccgc cagcccccgc
     2821 ccctgctggg tttgcaggtt tgggggcgtg acagttattg caggggttcg tgacagttat
     2881 tgcagggggg cgtgacagtt attgcagggg ttcgtgacag ttagtacggg agtgacgggc
     2941 actggctggc aatgtctagc aacggcaggc atttcggctg agggtaaaag aactttccgc
     3001 taagcgatag actgtatgta aacacagtat tgcaaggacg cggaacatgc ctcatgtggc
     3061 ggccaggacg gccagccggg atcgggatac tggtcgttac cagagccacc gacccgagca
     3121 aacccttctc tatcagatcg ttgacgagta ttacccggca ttcgctgcgc ttatggcaga
     3181 gcagggaaag gaattgccgg gctatgtgca acgggaattt gaagaatttc tccaatgcgg
     3241 gcggctggag catggctttc tacgggttcg ctgcgagtct tgccacgccg agcacctggt
     3301 cgctttcaga aatcaatcta aagtatatat gagtaaactt ggtctgacag gcccctgaat
     3361 tcgcatctag atggtagagc cacaaacagc cggtacaagc aacgatctcc aggaccatct
     3421 gaatcatgcg cggatgacac gaactcacga cggcgatcac agacattaac ccacagtaca
     3481 gacactgcga caacgtggca attcgtcgca ataccgtctc actgaactgg ccgataattg
     3541 cagacgaacg gacagtcatt catctttctg cccctccaaa agcaaaaacc cgccgaagcg
     3601 ggtttttacg taaatcaggt gaaactgacc gactttggca gtttattctt gacatgtagt
     3661 gagggggctg gtataatcac atagtactgt ttatgagaag agtaacttgt gaccttctct
     3721 cattcaaagg acaagttaat gacagtgctt gcgctgctaa ctgtctcagt ttgggtaaag
     3781 ctggaggtca ttgcgagaaa ggagtttgta tttgtcgaaa aaccagtttc aaagatctct
     3841 gggacaaacg tttcggttaa gtgccattat gaagattcaa tccaacagta ctatgtgatt
     3901 ataccagccc cctcactaca tgtcaagaat aaactgccaa aggtaatcgt taatccgcaa
     3961 ataacgtaaa aacccgcttc ggcgggtttt tttatggggg gagtttaggg aaagagcatt
     4021 tgtcagctga gcatgagacg gaaatctgct cgtcagtggt gctcacactg acgaatcatg
     4081 tacagatcat accgatgact gcctggcgac tcacaactaa gcaagacagc cggaaccagc
     4141 gccggcgaac accactgcat atatggcata tcacaacagt ccaactagtg cactgcagta
     4201 caccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     4261 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     4321 ctgcgtttat accgagcggc cgcttatttg ccgactacct tggtgatctc gcctttcacg
     4381 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
     4441 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     4501 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     4561 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     4621 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     4681 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     4741 gccagatcaa tgtcgatcgt ggctggctcg aagatacccg caagaatgtc attgcgctgc
     4801 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     4861 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     4921 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     4981 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     5041 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     5101 tgagtcgata cttcggcgat caccgcttcc ctcataatgt ttaactttgt tttagggcga
     5161 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     5221 cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa
     5281 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     5341 ttgcgtgagc gcatacgcta cttgcattac agcttacgaa ccgaacaggc ttatgtccac
     5401 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggtagcagc
     5461 gaagtcgagg catttctgtc ctggctggtc atgaccaaaa tcccttaacg tgagtcagcc
     5521 tgccgccttg ggccgggtga tgtcgtactt gcccgccgcg aactcggtta ccgtccagcc
     5581 cagcgcgacc agctccggca acgcctcgcg cacccgctgg cggcgcttgc gcatggtcga
     5641 accactggcc tctgacggcc agacatagcc gcacaaggta tctatggaag ccttgccggt
     5701 tttgccgggg tcgatccagc cacacagccg ctggtgcagc aggcgggcgg tttcgctgtc
     5761 cagcgcccgc acctcgtcca tgctgatgcg cacatgctgg ccgccaccca tgacggcctg
     5821 cgcgatcaag gggttcaggg ccacgtacag gcgcccgtcc gcctcgtcgc tggcgtactc
     5881 cgacagcagc cgaaacccct gccgcttgcg gccattctgg gcgatgatgg ataccttcca
     5941 aaggcgctcg atgcagtcct gtatgtgctt gagcgcccca ccactatcga cctctgcccc
     6001 gatttccttt gccagcgccc gatagctacc tttgaccacc atggcatcag cggtgacggc
     6061 ctcccacttg ggttccagga acagccggag ctgccgtccg ccttcggtct tgggttccgg
     6121 gccaagcact aggccattag gcccagccat ggccaccagc ccttgcagga tgcgcagatc
     6181 atcagcgccc agcggctccg ggccgctgaa ctcgatccgc ttgccgtcgc cgtagtcata
     6241 cgtcacgtcc agcttgctgc gcttgcgctc gccccgcttg agggcacgga acaggccggg
     6301 ggccagacag tgcgccgggt cgtgccggac gtggctgagg ctgtgcttgt tcttaggctt
     6361 caccacgggg cacccccttg ctcttgcgct gcctctccag cacggcgggc ttgagcaccc
     6421 cgccgtcatg ccgcctgaac caccgatcag cgaacggtgc gccatagttg gccttgctca
     6481 caccgaagcg gacgaagaac cggcgctggt cgtcgtccac accccattcc tcggcctcgg
     6541 cgctggtcat gctcgacagg taggactgcc agcggatgtt atcgaccagt accgagctgc
     6601 cccggctggc ctgctgctgg tcgcctgcgc ccatcatggc cgcgcccttg ctggcatggt
     6661 gcaggaacac gatagagcac ccggtatcgg cggcgatggc ctccatgcga ccgatgacct
     6721 gggccatggg gccgctggcg ttttcttcct cgatgtggaa ccggcgcagc gtgtccagca
     6781 ccatcaggcg gcggccctcg gcggcgcgct tgaggccgtc gaaccactcc ggggccatga
     6841 tgttgggcag gctgccgatc agcggctgga tcagcaggcc gtcagccacg gcttgccgtt
     6901 cctcggcgct gaggtgcgcc ccaagggcgt gcaggcggtg atgaatggcg gtgggcgggt
     6961 cttcggcggg caggtagatc accgggccgg tgggcagttc gcccacctcc agcagatccg
     7021 gcccgcctgc aatctgtgcg gccagttgca gggccagcat ggatttaccg gcaccaccgg
     7081 gcgacaccag cgccccgacc gtaccggcca ccatgttggg caaaacgtag tccagcggtg
     7141 gcggcgctgc tgcgaacgcc tccagaatat tgataggctt atgggtagcc attgattgcc
     7201 tcctttgcag gcagttggtg gttaggcgct ggcggggtca ctacccccgc cctgcgccgc
     7261 tctgagttct tccaggcact cgcgcagcgc ctcgtattcg tcgtcggtca gccagaactt
     7321 gcgctgacgc atccctttgg ccttcatgcg ctcggcatat cgcgcttggc gtacagcgtc
     7381 agggctggcc agcaggtcgc cggtctgctt gtccttttgg tctttcatat cagtcaccga
     7441 gaaacttgcc ggggccgaaa ggcttgtctt cgcggaacaa ggacaaggtg cagccgtcaa
     7501 ggttaaggct ggccatatca gcgactgaaa agcggccagc ctcggccttg tttgacgtat
     7561 aaccaaagcc accgggcaac caatagccct tgtcactttt gatcaggtag accgaccctg
     7621 aagcgctttt ttcgtattcc ataaaacccc cttctgtgcg tgagtactca tagtataaca
     7681 ggcgtgagta ccaacgcaag cactacatgc tgaaatctgg cccgcccctg tccatgcctc
     7741 gctggcgggg tgccggtgcc cgtgccagct cggcccgcgc aagctggacg ctgggcagac
     7801 ccatgacctt gctgacggtg cg
//
