LOCUS       sequence-448420-        4878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            E2-crimson, ampR and pBBR1 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4878)
  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 4878)
  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: pAC23
FEATURES             Location/Qualifiers
     source          1..4878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(135..800)
                     /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      801..1572
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      2823..2859
                     /label=soxR terminator
                     /note="bidirectional E. coli soxR transcription terminator"
     CDS             3013..3687
                     /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     3411..3430
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(join(3951..4742,4743..4811))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4574..4593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4812..4878,1..38))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
       61 cctgtcatga ccaaaatccc ttaacgtgac gcccggtagt gatcttattt cattatggtg
      121 aaagttggaa cctcttacgt acggcgcggc agcgtgaccc gtgtcggcgg ctccaacggc
      181 tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca ggcgctgctg cccgcgtcgt
      241 tcccattcct ccgtttcggt caaggctggc aggtctggtt ccatgcccgg aatgccgggc
      301 tggctgggcg gctcctcgcc ggggccggtc ggtagttgct gctcgcccgg atacagggtc
      361 gggatgcggc gcaggtcgcc atgccccaac agcgattcgt cctggtcgtc gtgatcaacc
      421 accacggcgg cactgaacac cgacaggcgc aactggtcgc ggggctggcc ccacgccacg
      481 cggtcattga ccacgtaggc cgacacggtg ccggggccgt tgagcttcac gacggagatc
      541 cagcgctcgg ccaccaagtc cttgactgcg tattggaccg tccgcaaaga acgtccgatg
      601 agcttggaaa gtgtcttctg gctgaccacc acggcgttct ggtggcccat ctgcgccacg
      661 aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa taagcccggc ccacgcctca
      721 tgcgctttgc gttccgtttg cacccagtga ccgggcttgt tcttggcttg aatgccgatt
      781 tctctggact gcgtggccat gcttatctcc atgcggtagg ggtgccgcac ggttgcggca
      841 ccatgcgcaa tcagctgcaa cttttcggca gcgcgacaac aattatgcgt tgcgtaaaag
      901 tggcagtcaa ttacagattt tctttaacct acgcaatgag ctattgcggg gggtgccgca
      961 atgagctgtt gcgtaccccc cttttttaag ttgttgattt ttaagtcttt cgcatttcgc
     1021 cctatatcta gttctttggt gcccaaagaa gggcacccct gcggggttcc cccacgcctt
     1081 cggcgcggct ccccctccgg caaaaagtgg cccctccggg gcttgttgat cgactgcgcg
     1141 gccttcggcc ttgcccaagg tggcgctgcc cccttggaac ccccgcactc gccgccgtga
     1201 ggctcggggg gcaggcgggc gggcttcgcc cttcgactgc ccccactcgc ataggcttgg
     1261 gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc
     1321 cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc
     1381 ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg
     1441 ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag
     1501 gcgcagcctg tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca
     1561 gccggtggcc gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga
     1621 ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc
     1681 acgactttgc gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc
     1741 accgccttgc gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc
     1801 ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct
     1861 caagcacgcc taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa
     1921 cgagcactgg gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc
     1981 agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga
     2041 atggtggaag tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg
     2101 gctggcggac aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac
     2161 cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa
     2221 ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt
     2281 ggccgatcta gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca
     2341 ggcgttctac gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc
     2401 ggtcgagcca cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg
     2461 cgttgagacg ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga
     2521 gcctgcccta caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca
     2581 agagacggcc cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt
     2641 gaatcgggat atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct
     2701 gacggaacag cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg
     2761 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgagcgga
     2821 acaaaactaa agcgccacaa gggcgcttta gtttgttttc agtccagtta cgctggagtc
     2881 tgaggctcgt catctgaatc atgcgcggat gacacgaact cacgacgctt tggcagttta
     2941 ttcttgacat gtagtgaggg ggctggtata atcacatagt actgttatac agaaacagag
     3001 gagatattac atatggatag cactgagaac gtcatcaagc ccttcatgcg cttcaaggtg
     3061 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgtggg cgagggcaag
     3121 ccctacgagg gcacccagac cgccaagctg caagtgacca agggcggccc cctgcccttc
     3181 gcctgggaca tcctgtcccc ccagttcttc tacggctcca aggcgtacat caagcacccc
     3241 gccgacatcc ccgactacct caagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg
     3301 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     3361 accctcatct accacgtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg
     3421 cagaagaaga ctctgggctg ggagccctcc actgagcgca actacccccg cgacggcgtg
     3481 ctgaagggcg agaaccacat ggcgctgaag ctgaagggcg gcggccacta cctgtgtgag
     3541 ttcaagtcca tctacatggc caagaagccc gtgaagctgc ccggctacca ctacgtggac
     3601 tacaagctcg acatcacctc ccacaacgag gactacaccg tggtggagca gtacgagcgc
     3661 gccgaggccc gccaccacct gttccagact cacggtatgg acgaattgta caagcacgac
     3721 gaattgtaaa tccgtaatcg ttaatccgca aataacgtaa aaacccgctt cggcgggttt
     3781 ttttatgggg ggagtttagg gaaagagcat ttgtcagctg gatcagcaga tgggaaccgg
     3841 atactcgagc ggccgcgatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4081 atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4861 atgagcggat acatattt
//
