LOCUS       sequence-398296-        8100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible two-plasmid system bearing CP25 > lacO-GFP, specR, 
            RSF1010 replicon (1/2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8100)
  AUTHORS   Chhun A, Moriano-Gutierrez S, Zoppi F, Cabirol A, Engel P, Schaerli 
            Y
  TITLE     An engineered bacterial symbiont allows noninvasive biosensing of 
            the honey bee gut environment.
  JOURNAL   PLoS Biol. 2024 Mar 5;22(3):e3002523. doi: 
            10.1371/journal.pbio.3002523. eCollection 2024 Mar.
  PUBMED    38442124
REFERENCE   2  (bases 1 to 8100)
  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"; journalName: "PLoS 
            Biol."; date: "2024-03-5"; volume: "22(3):e3002523. doi"; pages: " 
            10.1371/journal.pbio.3002523. eCollection 2024 Mar"
COMMENT     Sequence Label: pAC17V5a
FEATURES             Location/Qualifiers
     source          1..8100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(22..993)
                     /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            2232..2319
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      2661..3055
                     /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)"
     protein_bind    3812..3836
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             complement(4763..5554)
                     /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(5934..6785)
                     /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(6772..7611)
                     /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"
ORIGIN
        1 gcgtgagtac caacgcaagc actacatgct gaaatctggc ccgcccctgt ccatgcctcg
       61 ctggcggggt gccggtgccc gtgccagctc ggcccgcgca agctggacgc tgggcagacc
      121 catgaccttg ctgacggtgc gctcgatgta atccgcttcg tggccgggct tgcgctctgc
      181 cagcgctggg ctggcctcgg ccatggcctt gccgatttcc tcggcactgc ggccccggct
      241 ggccagcttc tgcgcggcga taaagtcgca cttgctgagg tcatcaccga agcgcttgac
      301 cagcccggcc atctcgctgc ggtactcgtc cagcgccgtg cgccggtggc ggctaagctg
      361 ccgctcgggc agttcgaggc tggccagcct gcgggccttc tcctgctgcc gctgggcctg
      421 ctcgatctgc tggccagcct gctgcaccag cgccgggcca gcggtggcgg tcttgccctt
      481 ggattcacgc agcagcaccc acggctgata accggcgcgg gtggtgtgct tgtccttgcg
      541 gttggtgaag cccgccaagc ggccatagtg gcggctgtcg gcgctggccg ggtcggcgtc
      601 gtactcgctg gccagcgtcc gggcaatctg cccccgaagt tcaccgcctg cggcgtcggc
      661 caccttgacc catgcctgat agttcttcgg gctggtttcc actaccaggg caggctcccg
      721 gccctcggct ttcatgtcat ccaggtcaaa ctcgctgagg tcgtccacca gcaccagacc
      781 atgccgctcc tgctcggcgg gcctgatata cacgtcattg ccctgggcat tcatccgctt
      841 gagccatggc gtgttctgga gcacttcggc ggctgaccat tcccggttca tcatctggcc
      901 ggtggtggcg tccctgacgc cgatatcgaa gcgctcacag cccatggcct tgagctgtcg
      961 gcctatggcc tgcaaagtcc tgtcgttctt catcgggcca ccaagcgcag ccagatcgag
     1021 ccgtcctcgg ttgtcagtgg cgtcaggtcg agcaagagca acgatgcgat cagcagcacc
     1081 accgtaggca tcatggaagc cagcatcacg gttagccata gcttccagtg ccacccccgc
     1141 gacgcgctcc gggcgctctg cgcggcgctg ctcacctcgg cggctacctc ccgcaactct
     1201 ttggccagct ccacccatgc cgcccctgtc tggcgctggg ctttcagcca ctccgccgcc
     1261 tgcgcctcgc tggcctgctg ggtctggctc atgacctgcc gggcttcgtc ggccagtgtc
     1321 gccatgctct gggccagcgg ttcgatctgc tccgctaact cgttgatgcc tctggatttc
     1381 ttcactctgt cgattgcgtt catggtctat tgcctcccgg tattcctgta agtcgatgat
     1441 ctgggcgttg gcggtgtcga tgttcagggc cacgtctgcc cggtcggtgc ggatgccccg
     1501 gccttccatc tccaccacgt tcggccccag gtgaacaccg ggcaggcgct cgatgccctg
     1561 cgcctcaagt gttctgtggt caatgcgggc gtcgtggcca gcccgctcta atgcccggtt
     1621 ggcatggtcg gcccatgcct cgcgggtctg ctcaagccat gccttgggct tgagcgcttc
     1681 ggtcttctgt gccccgccct tctccggggt cttgccgttg taccgcttga accactgagc
     1741 ggcgggccgc tcgatgccgt cattgatccg ctcggagatc atcaggtggc agtgcgggtt
     1801 ctcgccgcca ccggcatgga tggccagcgt atacggcagg cgctcggcac cggtcaggtg
     1861 ctgggcgaac tcggacgcca gcgccttctg ctggtcgagg gtcagctcga ccggcagggc
     1921 aaattcgacc tccttgaaca gccgcccatt ggcgcgttca tacaggtcgg cagcatccca
     1981 gtagtcggcg ggccgctcga cgaactccgg catgtgcccg gattcggcgt gcaagacttc
     2041 atccatgtcg cgggcatact tgccttcgcg ctggatgtag tcggccttgg ccctggccga
     2101 ttggccgccc gacctgctgc cggttttcgc cgtaaggtga taaatcgcca tgctgcctcg
     2161 ctgttgcttt tgcttttcgg ctccatgcaa tggccctcgg agagcgcacc gcccgaaggg
     2221 tggccgttag gccagtttct cgaagagaaa ccggtaagtg cgccctcccc tacaaagtag
     2281 ggtcgggatt gccgccgctg tgcctccatg atagcctacg agacagcaca ttaacaatgg
     2341 ggtgtcaaga tggttaaggg gagcaacaag gcggcggatc ggctggccaa gctcgaagaa
     2401 caacgagcgc gaatcaatgc cgaaattcag cgggtgcggg caagggaaca gcagcaagag
     2461 cgcaagaacg aaacaaggcg caaggtgctg gtgggggcca tgattttggc caaggtgaac
     2521 agcagcgagt ggccggagga tcggctcatg gcggcaatgg atgcgtacct tgaacgcgac
     2581 cacgaccgcg ccttgttcgg tctgccgcca cgccagaagg atgagccggg ctgaatgatc
     2641 gaccgagaca ggccctgcgg ggctgcacac gcgcccccac ccttcgggta gggggaaagg
     2701 ccgctaaagc ggctaaaagc gctccagcgt atttctgcgg ggtttggtgt ggggtttagc
     2761 gggctttgcc cgcctttccc cctgccgcgc agcggtgggg cggtgtgtag cctagcgcag
     2821 cgaatagacc agctatccgg cctctggccg ggcatattgg gcaagggcag cagcgcccca
     2881 caagggcgct gataaccgcg cctagtggat tattcttaga taatcatgga tggatttttc
     2941 caacaccccg ccagcccccg cccctgctgg gtttgcaggt ttgggggcgt gacagttatt
     3001 gcaggggttc gtgacagtta ttgcaggggg gcgtgacagt tattgcaggg gttcgtgaca
     3061 gttagtacgg gagtgacggg cactggctgg caatgtctag caacggcagg catttcggct
     3121 gagggtaaaa gaactttccg ctaagcgata gactgtatgt aaacacagta ttgcaaggac
     3181 gcggaacatg cctcatgtgg cggccaggac ggccagccgg gatcgggata ctggtcgtta
     3241 ccagagccac cgacccgagc aaacccttct ctatcagatc gttgacgagt attacccggc
     3301 attcgctgcg cttatggcag agcagggaaa ggaattgccg ggctatgtgc aacgggaatt
     3361 tgaagaattt ctccaatgcg ggcggctgga gcatggcttt ctacgggttc gctgcgagtc
     3421 ttgccacgcc gagcacctgg tcgctttcag aaatcaatct aaagtatata tgagtaaact
     3481 tggtctgaca ggcccctgaa ttcgcatcta gactgatgag acgtggtaga gccacaaaca
     3541 gccggtacaa gcaacgatct ccaggaccat ctgaatcatg cgcggatgac acgaactcac
     3601 gacggcgatc acagacatta acccacagta cagacactgc gacaacgtgg caattcgtcg
     3661 caatacaacg tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg
     3721 cttttggagg ggcagaaaga tgaatgactg tcctttggca gtttattctt gacatgtagt
     3781 gagggggctg gtataatcac atagtactgt tggaattgtg agcggataac aattccatac
     3841 agaaacagag gagatattac atatgagtaa aggagaagag cttttcacag gagttgtccc
     3901 aatcctcgtg gaattagacg gtgatgttaa tgggcacaag ttctctgtca gtggagaggg
     3961 tgaaggcgac gcaacatatg gcaagctgac ccttaaattt atttgcacca cgggtaaact
     4021 acctgttcca tggccaacac tggtcactac gttcgggtat ggggttcagt gctttgcgcg
     4081 ctacccagat cacatgaaac agcacgactt tttcaagagt gcaatgcccg aaggctatgt
     4141 acaggagaga accatctttt ttaaggatga cggcaactat aagacacgcg ccgaagtgaa
     4201 gttcgagggt gatacccttg ttaatagaat cgagttaaag ggtattgact ttaaggaaga
     4261 tggaaatatt ttaggccaca aactggaata taactataac tcccataatg tgtacattat
     4321 ggccgacaag caaaagaacg gtatcaaggt taacttcaag atcagacaca acattgagga
     4381 tggaagcgtt caactagccg accattacca acaaaacacc ccaattggcg atgggcctgt
     4441 gctgttacca gacaaccatt acctgtccac tcaatctgcc ctttcgaaag atcccaacga
     4501 aaagcgcgac cacatggtcc ttcttgagtt tgtcacggct gctgggatta cacacggcat
     4561 ggatgaacta tacaaataaa tccgtaatcg ttaatccgca aataacgtaa aaacccgctt
     4621 cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagctg cgtctcaagc
     4681 agttacagag atgttacgaa cccccaggac atccgagaat gcgaggcgat ggagggtaca
     4741 accgggagag tgttcacgcc gcttatttgc cgactacctt ggtgatctcg cctttcacgt
     4801 agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa
     4861 gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg
     4921 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc
     4981 gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata
     5041 gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt
     5101 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag
     5161 ccagatcaat gtcgatcgtg gctggctcga agatacccgc aagaatgtca ttgcgctgcc
     5221 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca
     5281 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
     5341 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
     5401 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
     5461 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
     5521 gagtcgatac ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac
     5581 tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
     5641 gcgcttgctg cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag
     5701 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
     5761 tgcgtgagcg catacgctac ttgcattaca gcttacgaac cgaacaggct tatgtccact
     5821 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggtagcagcg
     5881 aagtcgaggc atttctgtcc tggctggtca tgaccaaaat cccttaacgt gagtcagcct
     5941 gccgccttgg gccgggtgat gtcgtacttg cccgccgcga actcggttac cgtccagccc
     6001 agcgcgacca gctccggcaa cgcctcgcgc acccgctggc ggcgcttgcg catggtcgaa
     6061 ccactggcct ctgacggcca gacatagccg cacaaggtat ctatggaagc cttgccggtt
     6121 ttgccggggt cgatccagcc acacagccgc tggtgcagca ggcgggcggt ttcgctgtcc
     6181 agcgcccgca cctcgtccat gctgatgcgc acatgctggc cgccacccat gacggcctgc
     6241 gcgatcaagg ggttcagggc cacgtacagg cgcccgtccg cctcgtcgct ggcgtactcc
     6301 gacagcagcc gaaacccctg ccgcttgcgg ccattctggg cgatgatgga taccttccaa
     6361 aggcgctcga tgcagtcctg tatgtgcttg agcgccccac cactatcgac ctctgccccg
     6421 atttcctttg ccagcgcccg atagctacct ttgaccacca tggcatcagc ggtgacggcc
     6481 tcccacttgg gttccaggaa cagccggagc tgccgtccgc cttcggtctt gggttccggg
     6541 ccaagcacta ggccattagg cccagccatg gccaccagcc cttgcaggat gcgcagatca
     6601 tcagcgccca gcggctccgg gccgctgaac tcgatccgct tgccgtcgcc gtagtcatac
     6661 gtcacgtcca gcttgctgcg cttgcgctcg ccccgcttga gggcacggaa caggccgggg
     6721 gccagacagt gcgccgggtc gtgccggacg tggctgaggc tgtgcttgtt cttaggcttc
     6781 accacggggc acccccttgc tcttgcgctg cctctccagc acggcgggct tgagcacccc
     6841 gccgtcatgc cgcctgaacc accgatcagc gaacggtgcg ccatagttgg ccttgctcac
     6901 accgaagcgg acgaagaacc ggcgctggtc gtcgtccaca ccccattcct cggcctcggc
     6961 gctggtcatg ctcgacaggt aggactgcca gcggatgtta tcgaccagta ccgagctgcc
     7021 ccggctggcc tgctgctggt cgcctgcgcc catcatggcc gcgcccttgc tggcatggtg
     7081 caggaacacg atagagcacc cggtatcggc ggcgatggcc tccatgcgac cgatgacctg
     7141 ggccatgggg ccgctggcgt tttcttcctc gatgtggaac cggcgcagcg tgtccagcac
     7201 catcaggcgg cggccctcgg cggcgcgctt gaggccgtcg aaccactccg gggccatgat
     7261 gttgggcagg ctgccgatca gcggctggat cagcaggccg tcagccacgg cttgccgttc
     7321 ctcggcgctg aggtgcgccc caagggcgtg caggcggtga tgaatggcgg tgggcgggtc
     7381 ttcggcgggc aggtagatca ccgggccggt gggcagttcg cccacctcca gcagatccgg
     7441 cccgcctgca atctgtgcgg ccagttgcag ggccagcatg gatttaccgg caccaccggg
     7501 cgacaccagc gccccgaccg taccggccac catgttgggc aaaacgtagt ccagcggtgg
     7561 cggcgctgct gcgaacgcct ccagaatatt gataggctta tgggtagcca ttgattgcct
     7621 cctttgcagg cagttggtgg ttaggcgctg gcggggtcac tacccccgcc ctgcgccgct
     7681 ctgagttctt ccaggcactc gcgcagcgcc tcgtattcgt cgtcggtcag ccagaacttg
     7741 cgctgacgca tccctttggc cttcatgcgc tcggcatatc gcgcttggcg tacagcgtca
     7801 gggctggcca gcaggtcgcc ggtctgcttg tccttttggt ctttcatatc agtcaccgag
     7861 aaacttgccg gggccgaaag gcttgtcttc gcggaacaag gacaaggtgc agccgtcaag
     7921 gttaaggctg gccatatcag cgactgaaaa gcggccagcc tcggccttgt ttgacgtata
     7981 accaaagcca ccgggcaacc aatagccctt gtcacttttg atcaggtaga ccgaccctga
     8041 agcgcttttt tcgtattcca taaaaccccc ttctgtgcgt gagtactcat agtataacag
//
