LOCUS       sequence-388863-        5076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            E2-crimson, specR and pBBR1 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5076)
  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 5076)
  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: pAC04
FEATURES             Location/Qualifiers
     source          1..5076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(321..986)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRRT"
     rep_origin      987..1758
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      3009..3045
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     CDS             3308..3982
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     3706..3725
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(4182..4973)
                     /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"
ORIGIN
        1 gcccgaggca tagactgtac cccaaaaaaa cagtcataac aagccatgaa aaccgccact
       61 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
      121 tacttgcatt acagcttacg aaccgaacag gcttatgtcc actgggttcg tgccttcatc
      181 cgtttccacg gtgtgcgtca cccggcaacc ttgggtagca gcgaagtcga ggcatttctg
      241 tcctggctgg tcatgaccaa aatcccttaa cgtgacgccc ggtagtgatc ttatttcatt
      301 atggtgaaag ttggaacctc ttacgtacgg cgcggcagcg tgacccgtgt cggcggctcc
      361 aacggctcgc catcgtccag aaaacacggc tcatcgggca tcggcaggcg ctgctgcccg
      421 cgtcgttccc attcctccgt ttcggtcaag gctggcaggt ctggttccat gcccggaatg
      481 ccgggctggc tgggcggctc ctcgccgggg ccggtcggta gttgctgctc gcccggatac
      541 agggtcggga tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg gtcgtcgtga
      601 tcaaccacca cggcggcact gaacaccgac aggcgcaact ggtcgcgggg ctggccccac
      661 gccacgcggt cattgaccac gtaggccgac acggtgccgg ggccgttgag cttcacgacg
      721 gagatccagc gctcggccac caagtccttg actgcgtatt ggaccgtccg caaagaacgt
      781 ccgatgagct tggaaagtgt cttctggctg accaccacgg cgttctggtg gcccatctgc
      841 gccacgaggt gatgcagcag cattgccgcc gtgggtttcc tcgcaataag cccggcccac
      901 gcctcatgcg ctttgcgttc cgtttgcacc cagtgaccgg gcttgttctt ggcttgaatg
      961 ccgatttctc tggactgcgt ggccatgctt atctccatgc ggtaggggtg ccgcacggtt
     1021 gcggcaccat gcgcaatcag ctgcaacttt tcggcagcgc gacaacaatt atgcgttgcg
     1081 taaaagtggc agtcaattac agattttctt taacctacgc aatgagctat tgcggggggt
     1141 gccgcaatga gctgttgcgt accccccttt tttaagttgt tgatttttaa gtctttcgca
     1201 tttcgcccta tatctagttc tttggtgccc aaagaagggc acccctgcgg ggttccccca
     1261 cgccttcggc gcggctcccc ctccggcaaa aagtggcccc tccggggctt gttgatcgac
     1321 tgcgcggcct tcggccttgc ccaaggtggc gctgccccct tggaaccccc gcactcgccg
     1381 ccgtgaggct cggggggcag gcgggcgggc ttcgcccttc gactgccccc actcgcatag
     1441 gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc gcgagccttg
     1501 acccgccttc cacttggtgt ccaaccggca agcgaagcgc gcaggccgca ggccggaggc
     1561 ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc gcagttggag
     1621 ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca atccctgtgg tcaagctcgt
     1681 gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt gtccacgggc cgagcgaagc
     1741 gagccagccg gtggccgctc gcggccatcg tccacatatc cacgggctgg caagggagcg
     1801 cagcgaccgc gcagggcgaa gcccggagag caagcccgta gggcgccgca gccgccgtag
     1861 gcggtcacga ctttgcgaag caaagtctag tgagtatact caagcattga gtggcccgcc
     1921 ggaggcaccg ccttgcgctg cccccgtcga gccggttgga caccaaaagg gaggggcagg
     1981 catggcggca tacgcgatca tgcgatgcaa gaagctggcg aaaatgggca acgtggcggc
     2041 cagtctcaag cacgcctacc gcgagcgcga gacgcccaac gctgacgcca gcaggacgcc
     2101 agagaacgag cactgggcgg ccagcagcac cgatgaagcg atgggccgac tgcgcgagtt
     2161 gctgccagag aagcggcgca aggacgctgt gttggcggtc gagtacgtca tgacggccag
     2221 cccggaatgg tggaagtcgg ccagccaaga acagcaggcg gcgttcttcg agaaggcgca
     2281 caagtggctg gcggacaagt acggggcgga tcgcatcgtg acggccagca tccaccgtga
     2341 cgaaaccagc ccgcacatga ccgcgttcgt ggtgccgctg acgcaggacg gcaggctgtc
     2401 ggccaaggag ttcatcggca acaaagcgca gatgacccgc gaccagacca cgtttgcggc
     2461 cgctgtggcc gatctagggc tgcaacgggg catcgagggc agcaaggcac gtcacacgcg
     2521 cattcaggcg ttctacgagg ccctggagcg gccaccagtg ggccacgtca ccatcagccc
     2581 gcaagcggtc gagccacgcg cctatgcacc gcagggattg gccgaaaagc tgggaatctc
     2641 aaagcgcgtt gagacgccgg aagccgtggc cgaccggctg acaaaagcgg ttcggcaggg
     2701 gtatgagcct gccctacagg ccgccgcagg agcgcgtgag atgcgcaaga aggccgatca
     2761 agcccaagag acggcccgag accttcggga gcgcctgaag cccgttctgg acgccctggg
     2821 gccgttgaat cgggatatgc aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa
     2881 gctgctgacg gaacagcggg aagtccagcg ccagaaacag gcccagcgcc agcaggaacg
     2941 cgggcgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     3001 agcggaacaa aactaaagcg ccacaagggc gctttagttt gttttcagtc cagttacgct
     3061 ggagtctgag gctcgtcatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca
     3121 cagacattaa cccacagtac agacactgcg acaacgtggc aattcgtcgc aataccgtct
     3181 cactgaactg gccgataatt gcagacgaac gggtgaaaca aaacggttga caacatgaag
     3241 taaacacggt acgatgtacc acatgaaacg acagtgagtc aatacagaaa cagaggagat
     3301 attacatatg gatagcactg agaacgtcat caagcccttc atgcgcttca aggtgcacat
     3361 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gtgggcgagg gcaagcccta
     3421 cgagggcacc cagaccgcca agctgcaagt gaccaagggc ggccccctgc ccttcgcctg
     3481 ggacatcctg tccccccagt tcttctacgg ctccaaggcg tacatcaagc accccgccga
     3541 catccccgac tacctcaagc agtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     3601 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcaccct
     3661 catctaccac gtgaagttca tcggcgtgaa cttcccctcc gacggccccg taatgcagaa
     3721 gaagactctg ggctgggagc cctccactga gcgcaactac ccccgcgacg gcgtgctgaa
     3781 gggcgagaac cacatggcgc tgaagctgaa gggcggcggc cactacctgt gtgagttcaa
     3841 gtccatctac atggccaaga agcccgtgaa gctgcccggc taccactacg tggactacaa
     3901 gctcgacatc acctcccaca acgaggacta caccgtggtg gagcagtacg agcgcgccga
     3961 ggcccgccac cacctgttcc agactcacgg tatggacgaa ttgtacaagc acgacgaatt
     4021 gtaaatccgt aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta
     4081 tggggggagt ttagggaaag agcatttgtc agctggatca gcagatggga accggatact
     4141 cctcagcctc cagagtagcc aatgagacgc cgagcggccg cttatttgcc gactaccttg
     4201 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     4261 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     4321 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     4381 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     4441 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     4501 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     4561 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacccgca
     4621 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     4681 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     4741 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     4801 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     4861 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     4921 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct cataatgttt
     4981 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
     5041 catcgaccca cggcgtaacg cgcttgctgc ttggat
//
