LOCUS       sequence-442535-        4471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            E2-crimson, ampR and RK2 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4471)
  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 4471)
  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: pAC25
FEATURES             Location/Qualifiers
     source          1..4471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            51..159
                     /note="incP origin of transfer"
     CDS             complement(220..1368)
                     /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      1761..2292
                     /label=oriV
                     /note="incP origin of replication"
     CDS             2520..3194
                     /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     2918..2937
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(join(3458..4249,4250..4318))
                     /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     4081..4100
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4319..4423)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
       61 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
      121 gcggtgctca acgggaatcc tgctctgcga ggctggccgt aggccggccg cgatgcaggt
      181 ggctgctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag
      241 gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc
      301 gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa
      361 ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg
      421 ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc
      481 aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg
      541 gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga
      601 cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata
      661 ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg
      721 ctcgccgata ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc
      781 gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat
      841 gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga
      901 gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa
      961 ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc
     1021 ttcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt
     1081 tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagacgacg
     1141 cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc
     1201 gcgctcgatc ttggccgtag cttgctggac tatcgagccg acggactgga aggtttcgcg
     1261 gggcgcacgc atgacggtgc ggcttacgat ggtttcggca tcctcggcgg aaaaccccgc
     1321 gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg
     1381 cgggattgcc ccggaattaa ttccccggat cgatccgtcg atcttgatcc cctgcgccat
     1441 cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca
     1501 gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc
     1561 tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt
     1621 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     1681 ttctacgtgg ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg
     1741 ggggatggga ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt
     1801 cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat
     1861 tggtttaaaa gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt
     1921 gcaaatgctg gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca
     1981 tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc
     2041 gcccctcaag tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg
     2101 gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc
     2161 cggccgaaat cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca
     2221 agtgtcaacg tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca
     2281 acgccggcgg ccctacatgg ctctgctgta gtgagtgggt tgcgctccgg cagcggtcct
     2341 gatcccccgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggcgcgcat
     2401 ctgaatcatg cgcggatgac acgaactcac gacgctttgg cagtttattc ttgacatgta
     2461 gtgagggggc tggtataatc acatagtact gttatacaga aacagaggag atattacata
     2521 tggatagcac tgagaacgtc atcaagccct tcatgcgctt caaggtgcac atggagggct
     2581 ccgtgaacgg ccacgagttc gagatcgagg gcgtgggcga gggcaagccc tacgagggca
     2641 cccagaccgc caagctgcaa gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     2701 tgtcccccca gttcttctac ggctccaagg cgtacatcaa gcaccccgcc gacatccccg
     2761 actacctcaa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2821 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcacc ctcatctacc
     2881 acgtgaagtt catcggcgtg aacttcccct ccgacggccc cgtaatgcag aagaagactc
     2941 tgggctggga gccctccact gagcgcaact acccccgcga cggcgtgctg aagggcgaga
     3001 accacatggc gctgaagctg aagggcggcg gccactacct gtgtgagttc aagtccatct
     3061 acatggccaa gaagcccgtg aagctgcccg gctaccacta cgtggactac aagctcgaca
     3121 tcacctccca caacgaggac tacaccgtgg tggagcagta cgagcgcgcc gaggcccgcc
     3181 accacctgtt ccagactcac ggtatggacg aattgtacaa gcacgacgaa ttgtaaatcc
     3241 gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg cgggtttttt tatgggggga
     3301 gtttagggaa agagcatttg tcagctggat cagcagatgg gaaccggata ctcgagcggc
     3361 cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3421 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3481 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3541 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg
     3601 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3661 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3721 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3781 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3841 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3901 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3961 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4021 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4081 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4141 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4201 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4261 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4321 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4381 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4441 tgccacctgt catgaccaaa atcccttaac g
//
