LOCUS       sequence-442531-        8299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            E2-crimson, ampR and pTFC-FC2 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8299)
  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 8299)
  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: pAC24
FEATURES             Location/Qualifiers
     source          1..8299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2230..2241)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             2448..2456
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             6351..7025
                     /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     6749..6768
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(join(7289..8080,8081..8149))
                     /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     7912..7931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8150..8254)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtgtcaag gcagccagaa catccaaaaa tccgctgtag gcgattctga tggcttctgt
       61 tcggggtatg aggcgatgtg ctccacaagc acgaccgaca agccagccga aacgctcgcc
      121 aagctggccg ctgcgctacc tgccggagag gccaaacgct accgggtgat ggtgtccatt
      181 gtcccggtgg aagaatgaag gcgagcgggg cggggcgggg tgtgtctggc cgggcttgtc
      241 ccggccttgt cttttggggt ttttggtaag gacttttgac gccctcaagg gcggcgaaaa
      301 gtaccgcgcc tagtggatga tcttggatag atcatggagg gattttgcga atcgcccgca
      361 aacccgcgcc gttgctaggt ttttcagatt tcggcgggga acgttacgcc gggggttggg
      421 gaacgttacg ccgggggttg gggaacgtta cgccgggggt tgggcctgct gatttcccac
      481 ttgcccttgg cgtactcgtt caccgtccaa ccaacggcgg caagctcgac aagcgccttg
      541 cgggcggtct ggcggcgctt tttcattgct tcatcgttgg ctgcgtctgg ccatacataa
      601 ccgcacagcg tgtcgatttc agctttgcca gactttccgg ggtcaatcca gccacatagc
      661 cgctggtgta gcagccgtgc cgggtctgtt tccagcttgc gaacttctgc catgttgatg
      721 cgggtgtgcg ggcgctctcc cataaccgca tccgccagcc ggggattaag cgcaacaaat
      781 aacttgcctt cttgctcgtc gctcgcgtac tcggacagaa tgcggaaccc ttgccgctta
      841 ccacctcttt cgacaatcac cgataccgcc caaagccgtt cgatgctttc ccggatggtt
      901 ttaaactgac ttcctccatc ttcggcgtag ccaagttctc gggccaactg gcggaagctg
      961 cccttggcta ccatcgtatc tttctctatg gcgtcccacc gcatatcaag ccataggcgg
     1021 agttgctggc ctgcttctga ttccgtttcg tgccgtagca cgatgccgcg cccgttatct
     1081 ccggaaattg cagccattgc caccagccct tgcaatacgc gcaagtcatc gccgccaagt
     1141 ggttcaggcc cccaaaaacg gattgagtca tctccgtagt tgtaggtcac atcgagcttg
     1201 agcctctttc gttctccgcg cttgagactg cggaaaagcc ccggcgtgag gcaatgcgcg
     1261 gggtcgtgcc gcgcatgggt gaggtcatag ctcattgcgc tttccccctt tttttcctgc
     1321 tccattgctg attgcctcct gtagttcaac gggcaacaac accccgccgc tgtgccgctg
     1381 ataccagcgg tctagcgggg tcgcgtcgta gttctgcttg ctcacgccaa agcggacaaa
     1441 aaggccgcgc cgctcgttgc cgatgggcga acgatcaaag ccccggtcac tcatgcgctc
     1501 ggcttcctgc tccgtcattt tggcgacata gccgcaccat ctggcgttgt cgatcagggc
     1561 agacgcgccc cgcgctgcct gctgctggtc ggtctggccg tcgcgggcgc tgcccttgtt
     1621 gacgtggtgc aggtacagga cagccgcccc ggtggtcgcc gcgatgtgtt ccaacacgga
     1681 aacaaggtgg gccatgtcgc cattgctgtt ctcgtcgagg ctgtgaatcc ggctcagggt
     1741 gtccagcaca atcagccggg cccctgcgct gtagtcgatg acgcggcgca agtgcgcctc
     1801 gtccatcacg tttagccgct tgcccatgat cggctcaagc atcaggttct cagcgatggc
     1861 ttcgcgggcc gactggttga ggtgctggcc gatggcgtgg acacgtcgca caagggcggg
     1921 ctgtggatct tcgccagcga gataaaccac gcgcccggtg tgcgccgggg ttagtcccac
     1981 aaggtcgccg cctgccacac tgcaagcgat tgacatggcc gcttcaagag caaaaaagct
     2041 cttgccagtt gcgccaggtg cgacaagtgc gcccacggtt ccggccaaga atccgggcca
     2101 gatgaaatca agttctggcg gctcattggt gaaggccgcc ataatgtcta aagccatgct
     2161 tgtctccttt tcagacagtt gtggttaggc gctgccgggg ccgcaatccc ctgccgcgcc
     2221 gctcaatttt cgcccctcga tgccagccac gccgaagcct cggccaaggc gtcctcatag
     2281 gcgtagctgg ccgcttcttc tgcgttctct gctgtggcgt actcggacaa cgggaacact
     2341 ccgccgccga cttcctcgcc gctttccaac aggcgcaacc gccagccgcc gccaaggtcg
     2401 gaggggcgcg ggtcgatctg gtagctgtat tcgatcattg gcggtaaacc tccttacgat
     2461 tgccgatgcg cataacgagg atgcgcaaag caccgtcctc gatgctggcg atgattcgcc
     2521 aatccccaac gcggtatttc cagaaggctc ccagtttgga gcctttgagg gcttcgccaa
     2581 tgctgcgcgg gtcgtcgagc tgggcgacac ggccatgcaa aaaagcgagg atgcggcggg
     2641 cggtctgctg gtcgatcttg cctagctcgc gctcggcggc tgggtcgagt tcaaccttcc
     2701 atgccatagc gtttcatcac ttcttcgagt ggcacggttt gggttttgcc tgcgcgaatg
     2761 tcgatcaggc gttgctctgc aaggtacaaa tcttcgaggt catccaagtg ctcaaggatg
     2821 gcttcgcggg catagaaagt cttggtccgc cctgtggcct gcgccagtgc ttcaaggcgg
     2881 gtttccactt cggcgggcag tcggattgca agcatgtttc ggctcctgtt ctgaaaacgc
     2941 tatacaagta tatcacaacc accacgcaac gctagagact cattcccggc gagttgtcgc
     3001 gctcgatctt ctgcgccttt gcctgctcct gctgtcgtac aggctcgcgc ccttccagct
     3061 tctcccactg ctgccggtac ttgccaagat cggcggcttg ccggtcgcct gcggcgctgt
     3121 aggcatagcg ggcggtgcgc tgcgcgtagt cgttccaatc gcggccctcg tctttctgcc
     3181 gggtggccgg tgcgcactgg cggatagcgc cctcgatggc cgcttgatcg tggccggtga
     3241 cgcgcatacg cacggcaatc atggaatcca cgcgggacaa gtccacctcg ccgccacgct
     3301 gccgcttgag cacgtcgcgg taatgccgct ggtaggcgtc gatcgcgctg ccgctggccg
     3361 ctgcaagctc caaggcgggc ttggctttgc tgcgctcggg ctgctgcgcc ttcaaggccg
     3421 cttgccgctg atactcggcg tcgatctggc tggacaaggc cagcgccttg acgcactccc
     3481 ggcgctcggc cttgagcaag cgcacctctg gatagctgcc gtcctcccgc tggtgcttgg
     3541 gcttgcggtt ctcgtagccg ggtgcgcggt gcgggtggat ggctcccgat agcttcgggt
     3601 cgccgtattc acggttcagg gcatcgctca ggcggttgcc cacgtcctta tcgtgggccg
     3661 tccccagctt cgacacggtg atgatggcct gatagttgcc ggggctggat tccagcacgg
     3721 cggcgggctg gtagccgtct ttgataagcc gctccagctt ctcccggttc atgtcgtcga
     3781 tgaggatgtg atgcttcttg tccgatagcg gcgtgtagta gaggttttcg ccccggcgct
     3841 gtaggcgctg catctccggc gtgcgctgct cgatctcctg cggtgtgaaa ccccgcgtga
     3901 tgccgtcctt cttgtcgagg atgaaggttt gcttccttcc gtctgcctgc atcttgatgg
     3961 acgttacccg gtagcgctcc gcgcctaccg cttccgcgta ccgctcgaac tgctttagct
     4021 gctccgattt catcgcctgc gccctctcct gctgtatccg ctgccgctcc tgctggatgc
     4081 tctcctgaag ctctgcattt gtgatcttga agccgtgctc tgctgctaac ttcccgcaca
     4141 tggccttgta ttcgtcgttc ccggttacgg tgaagctgcc ccacttctgc gccgatagct
     4201 gcaaggcggc aagcgtgctg tcccggttgc gccagtcgtg aatatcgatg cttttgccct
     4261 tgtccacgaa agacacaccc ccgccccgcc ccccttcttc cttccggctg taatgcacct
     4321 gctggcccac gatttcaggc tgataggcgc gaatgtcgcg cggcgtcggc ggctcgctgc
     4381 ggtcgccctc gatgcgctgc ggctcgctgg cccggtgtcg ccactgctcg gcaagctccg
     4441 ggcttttctg catccgctgc cactgctcca ggtcgggata gggtctgaac tgctgccggt
     4501 gctgctcgcg ctgcttctgg tgcttctcct tcaaggcggc tttctctgcg gcctgctcgg
     4561 cggcaatcac gctgcgcatg gcgttgagca cttcgccctt gcctttccag ttgccgcgca
     4621 tgagttcatc gcggcgggcc ttctgctgct cggcaagttg cttgcgctcc tgctcctgcc
     4681 gcttgtcctg taccagcttg tccgcgttct tctccgagta gtgcgccttg cgcccggtga
     4741 tgtagtcctt ccatcccggc acgtctggct tgatcggctc cggctcgcgc tgtgctacct
     4801 gctgacgctg cggcggctgg taagctccga gtcgcttttg caacttggat aggctcgcgt
     4861 cccggtctgc gctgctggcc ttcacgccaa catcaccgac aaagagcgtc gccccgctgc
     4921 ctgttttctc gtagcgcatc cccttggctg ctaactcgcg gtgcaactgc tcccacgtct
     4981 gcgccttttt gatgatcggt gcgccgtcct caatggcgat gcgttcggcg gatttctccc
     5041 cggtgcggtt ttccatgtcg cgcttcggct gggctggctg gcgcggcttg tccttgtcga
     5101 tgtgctcccg gcctaactca ccattctcca acacctgata acgcccgttc tgctcacgct
     5161 gccagccttg tgcgttctcg attcgtgcta tcgccttgtg cgctgcctca atgtcgaagc
     5221 cgttattgat cttcaccacc ttgagcgttt caggatgcac gcggttgatg gcaaggtgca
     5281 ggtgcaagtt gtccgtgtcg gcgtgcaaac catagatggc ttgatggtct ttaacaccta
     5341 gctcatccat gaaaatgctt accgcttctt cgacctgctc cgggctgggc tgctcgcctt
     5401 ctcgccaact caggacatag tgattgatcg tatccttgct tcgcactgcc tcttgtgaca
     5461 aggcaatcat ttctgccgtc tggctctgtg gttcatcggt gatgaatcca cgcgccccgg
     5521 catggatgca cttctcctgc gagttctcac gctcaggctc acggatgtag ccagttaggc
     5581 gggaaacgcg aaccgccttg cttgctgcct tctgcgggtt cttaaccttc ttgacgatca
     5641 cggctcagtt tcccgatata ggctttcaag accacaagcg ccccggctgt gtccttgctg
     5701 tacacgcctc cgctctcgtt gtggatgtgc ttaagcaatc caccgatgcg gcgcaactct
     5761 ttgagcatta ccgcgtcggc gctggcaatg atcggcctgc caaagtagcg gcggcggaca
     5821 agctccgaca tgctcaaccc ggcaaggtcg gcatcctctt tcaagcgggc cttctccgct
     5881 gcggtcagcc gtacattgat aacggcgtcc agcggttccg gcccctgcgt ttcaaatggc
     5941 atcgtgtctc cttctcgcgg ggggcgggcg gggtgtgtct tggacagtaa gaagcgagag
     6001 cgccgcactg tccggggtca aggggatacc ccttgcagga taagccagac cgtgagaagc
     6061 gaagcgccga atgggcaggc gtccgttgga accacgtggc gaagccatgt gtgttacaac
     6121 ggtcatcctg tattgctcaa ccgctctact atcatatcag cagtttaggc acactagatg
     6181 cactttgcaa caagcataaa aggcgctatg ctgtcaactc cgaaaccaga tctgaatcat
     6241 gcgcggatga cacgaactca cgacgctttg gcagtttatt cttgacatgt agtgaggggg
     6301 ctggtataat cacatagtac tgttatacag aaacagagga gatattacat atggatagca
     6361 ctgagaacgt catcaagccc ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     6421 gccacgagtt cgagatcgag ggcgtgggcg agggcaagcc ctacgagggc acccagaccg
     6481 ccaagctgca agtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     6541 agttcttcta cggctccaag gcgtacatca agcaccccgc cgacatcccc gactacctca
     6601 agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     6661 tggtgaccgt gacccaggac tcctccctgc aggacggcac cctcatctac cacgtgaagt
     6721 tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca gaagaagact ctgggctggg
     6781 agccctccac tgagcgcaac tacccccgcg acggcgtgct gaagggcgag aaccacatgg
     6841 cgctgaagct gaagggcggc ggccactacc tgtgtgagtt caagtccatc tacatggcca
     6901 agaagcccgt gaagctgccc ggctaccact acgtggacta caagctcgac atcacctccc
     6961 acaacgagga ctacaccgtg gtggagcagt acgagcgcgc cgaggcccgc caccacctgt
     7021 tccagactca cggtatggac gaattgtaca agcacgacga attgtaaatc cgtaatcgtt
     7081 aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga
     7141 aagagcattt gtcagctgga tcagcagatg ggaaccggat actcgagcgg ccgcgattat
     7201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg gacccacgct
     7441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8161 ttcaatatta ttgaagcatt tatcagagtt attgtctcat gagcggatac atatttgaat
     8221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8281 tcatgaccaa aatccctta
//
