LOCUS       sequence-388858-        4669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            E2-crimson, specR and RK2 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4669)
  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 4669)
  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: pAC06
FEATURES             Location/Qualifiers
     source          1..4669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            722..830
                     /note="incP origin of transfer"
     CDS             complement(891..2039)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIVSRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      2432..2963
                     /label=oriV
                     /note="incP origin of replication"
     CDS             3300..3974
                     /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     3698..3717
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(join(4174..4669,1..296))
                     /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 cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc
       61 tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca
      121 tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca
      181 tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg
      241 acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcataatg
      301 tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc
      361 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacc
      421 ccaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
      481 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagcttacga
      541 accgaacagg cttatgtcca ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac
      601 ccggcaacct tgggtagcag cgaagtcgag gcatttctgt cctggctggt catgaccaaa
      661 atcccttaac gtgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
      721 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
      781 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc
      841 gcgatgcagg tggctgctga acccccagcc ggaactgacc ccacaaggcc ctagcgtttg
      901 caatgcacca ggtcatcatt gacccaggcg tgttccacca ggccgctgcc tcgcaactct
      961 tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc cgatccgcac
     1021 atgaggcgga aggtttccag cttgagcggg tacggctccc ggtgcgagct gaaatagtcg
     1081 aacatccgtc gggccgtcgg cgacagcttg cggtacttct cccatatgaa tttcgtgtag
     1141 tggtcgccag caaacagcac gacgatttcc tcgtcgatca ggacctggca acgggacgtt
     1201 ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg acaccgattc caggtgccca
     1261 acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca ttcctcggcc
     1321 ttcgtgtaat accggccatt gatcgaccag cccaggtcct ggcaaagctc gtagaacgtg
     1381 aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg ctgccacacc
     1441 agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac gtccttgttg
     1501 acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt gcgcgtggtg
     1561 aacagggcag agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg ccacggcgca
     1621 atatcgaaca aggaaagctg catttccttg atctgctgct tcgtgtgttt cagcaacgcg
     1681 gcctgcttgg cttcgctgac ctgttttgcc aggtcctcgc cggcggtttt tcgcttcttg
     1741 gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc cgcctcctgt
     1801 tcgagacgac gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg gggagccagt
     1861 tgcacgctgt cgcgctcgat cttggccgta gcttgctgga ctatcgaacc gacggactgg
     1921 aaggtttcgc ggggcgcacg catgacggtg cggcttacga tggtttcggc atcctcggcg
     1981 gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt ccgattcatt
     2041 caccctcctt gcgggattgc cccggaatta attccccgga tcgatccgtc gatcttgatc
     2101 ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc
     2161 caaccttacc agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg
     2221 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
     2281 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     2341 gcggactggc tttctacgtg gctgccattt ttggggtgag gccgttcgcg gccgaggggc
     2401 gcagcccctg gggggatggg aggcccgcgt tagcgggccg ggagggttcg agaagggggg
     2461 gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg
     2521 tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg
     2581 cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg
     2641 tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg
     2701 tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac
     2761 atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag
     2821 ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag
     2881 tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg
     2941 aggtatccac aacgccggcg gccctacatg gctctgctgt agtgagtggg ttgcgctccg
     3001 gcagcggtcc tgatcccccg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3061 acggcgcgca tctgaatcat gcgcggatga cacgaactca cgacggcgat cacagacatt
     3121 aacccacagt acagacactg cgacaacgtg gcaattcgtc gcaataccgt ctcactgaac
     3181 tggccgataa ttgcagacga acgggtgaaa caaaacggtt gacaacatga agtaaacacg
     3241 gtacgatgta ccacatgaaa cgacagtgag tcaatacaga aacagaggag atattacata
     3301 tggatagcac tgagaacgtc atcaagccct tcatgcgctt caaggtgcac atggagggct
     3361 ccgtgaacgg ccacgagttc gagatcgagg gcgtgggcga gggcaagccc tacgagggca
     3421 cccagaccgc caagctgcaa gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     3481 tgtcccccca gttcttctac ggctccaagg cgtacatcaa gcaccccgcc gacatccccg
     3541 actacctcaa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3601 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcacc ctcatctacc
     3661 acgtgaagtt catcggcgtg aacttcccct ccgacggccc cgtaatgcag aagaagactc
     3721 tgggctggga gccctccact gagcgcaact acccccgcga cggcgtgctg aagggcgaga
     3781 accacatggc gctgaagctg aagggcggcg gccactacct gtgtgagttc aagtccatct
     3841 acatggccaa gaagcccgtg aagctgcccg gctaccacta cgtggactac aagctcgaca
     3901 tcacctccca caacgaggac tacaccgtgg tggagcagta cgagcgcgcc gaggcccgcc
     3961 accacctgtt ccagactcac ggtatggacg aattgtacaa gcacgacgaa ttgtaaatcc
     4021 gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg cgggtttttt tatgggggga
     4081 gtttagggaa agagcatttg tcagctggat cagcagatgg gaaccggata ctcctcagcc
     4141 tccagagtag ccaatgagac gccgagcggc cgcttatttg ccgactacct tggtgatctc
     4201 gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
     4261 ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag
     4321 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     4381 gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg
     4441 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     4501 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     4561 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacccg caagaatgtc
     4621 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgc
//
