LOCUS       sequence-388862-        8498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            E2-crimson, specR and pTFC-FC2 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8498)
  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 8498)
  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: pAC11
FEATURES             Location/Qualifiers
     source          1..8498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             231..905
                     /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     629..648
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(1105..1896)
                     /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"
     CDS             complement(4499..4510)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             4717..4725
                     /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)"
ORIGIN
        1 atctgaatca tgcgcggatg acacgaactc acgacggcga tcacagacat taacccacag
       61 tacagacact gcgacaacgt ggcaattcgt cgcaataccg tctcactgaa ctggccgata
      121 attgcagacg aacgggtgaa acaaaacggt tgacaacatg aagtaaacac ggtacgatgt
      181 accacatgaa acgacagtga gtcaatacag aaacagagga gatattacat atggatagca
      241 ctgagaacgt catcaagccc ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
      301 gccacgagtt cgagatcgag ggcgtgggcg agggcaagcc ctacgagggc acccagaccg
      361 ccaagctgca agtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
      421 agttcttcta cggctccaag gcgtacatca agcaccccgc cgacatcccc gactacctca
      481 agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
      541 tggtgaccgt gacccaggac tcctccctgc aggacggcac cctcatctac cacgtgaagt
      601 tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca gaagaagact ctgggctggg
      661 agccctccac tgagcgcaac tacccccgcg acggcgtgct gaagggcgag aaccacatgg
      721 cgctgaagct gaagggcggc ggccactacc tgtgtgagtt caagtccatc tacatggcca
      781 agaagcccgt gaagctgccc ggctaccact acgtggacta caagctcgac atcacctccc
      841 acaacgagga ctacaccgtg gtggagcagt acgagcgcgc cgaggcccgc caccacctgt
      901 tccagactca cggtatggac gaattgtaca agcacgacga attgtaaatc cgtaatcgtt
      961 aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga
     1021 aagagcattt gtcagctgga tcagcagatg ggaaccggat actcctcagc ctccagagta
     1081 gccaatgaga cgccgagcgg ccgcttattt gccgactacc ttggtgatct cgcctttcac
     1141 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     1201 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     1261 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     1321 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     1381 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     1441 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     1501 agccagatca atgtcgatcg tggctggctc gaagataccc gcaagaatgt cattgcgctg
     1561 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     1621 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     1681 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     1741 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     1801 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     1861 ttgagtcgat acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg
     1921 actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     1981 acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca
     2041 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     2101 gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca
     2161 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag
     2221 cgaagtcgag gcatttctgt cctggctggt catgaccaaa atcccttaac gtgctcaagg
     2281 cagccagaac atccaaaaat ccgctgtagg cgattctgat ggcttctgtt cggggtatga
     2341 ggcgatgtgc tccacaagca cgaccgacaa gccagccgaa acgctcgcca agctggccgc
     2401 tgcgctacct gccggagagg ccaaacgcta ccgggtgatg gtgtccattg tcccggtgga
     2461 agaatgaagg cgagcggggc ggggcggggt gtgtctggcc gggcttgtcc cggccttgtc
     2521 ttttggggtt tttggtaagg acttttgacg ccctcaaggg cggcgaaaag taccgcgcct
     2581 agtggatgat cttggataga tcatggaggg attttgcgaa tcgcccgcaa acccgcgccg
     2641 ttgctaggtt tttcagattt cggcggggaa cgttacgccg ggggttgggg aacgttacgc
     2701 cgggggttgg ggaacgttac gccgggggtt gggcctgctg atttcccact tgcccttggc
     2761 gtactcgttc accgtccaac caacggcggc aagctcgaca agcgccttgc gggcggtctg
     2821 gcggcgcttt ttcattgctt catcgttggc tgcgtctggc catacataac cgcacagcgt
     2881 gtcgatttca gctttgccag actttccggg gtcaatccag ccacatagcc gctggtgtag
     2941 cagccgtgcc gggtctgttt ccagcttgcg aacttctgcc atgttgatgc gggtgtgcgg
     3001 gcgctctccc ataaccgcat ccgccagccg gggattaagc gcaacaaata acttgccttc
     3061 ttgctcgtcg ctcgcgtact cggacagaat gcggaaccct tgccgcttac cacctctttc
     3121 gacaatcacc gataccgccc aaagccgttc gatgctttcc cggatggttt taaactgact
     3181 tcctccatct tcggcgtagc caagttctcg ggccaactgg cggaagctgc ccttggctac
     3241 catcgtatct ttctctatgg cgtcccaccg catatcaagc cataggcgga gttgctggcc
     3301 tgcttctgat tccgtttcgt gccgtagcac gatgccgcgc ccgttatctc cggaaattgc
     3361 agccattgcc accagccctt gcaatacgcg caagtcatcg ccgccaagtg gttcaggccc
     3421 ccaaaaacgg attgagtcat ctccgtagtt gtaggtcaca tcgagcttga gcctctttcg
     3481 ttctccgcgc ttgagactgc ggaaaagccc cggcgtgagg caatgcgcgg ggtcgtgccg
     3541 cgcatgggtg aggtcatagc tcattgcgct ttcccccttt ttttcctgct ccattgctga
     3601 ttgcctcctg tagttcaacg ggcaacaaca ccccgccgct gtgccgctga taccagcggt
     3661 ctagcggggt cgcgtcgtag ttctgcttgc tcacgccaaa gcggacaaaa aggccgcgcc
     3721 gctcgttgcc gatgggcgaa cgatcaaagc cccggtcact catgcgctcg gcttcctgct
     3781 ccgtcatttt ggcgacatag ccgcaccatc tggcgttgtc gatcagggca gacgcgcccc
     3841 gcgctgcctg ctgctggtcg gtctggccgt cgcgggcgct gcccttgttg acgtggtgca
     3901 ggtacaggac agccgccccg gtggtcgccg cgatgtgttc caacacggaa acaaggtggg
     3961 ccatgtcgcc attgctgttc tcgtcgaggc tgtgaatccg gctcagggtg tccagcacaa
     4021 tcagccgggc ccctgcgctg tagtcgatga cgcggcgcaa gtgcgcctcg tccatcacgt
     4081 ttagccgctt gcccatgatc ggctcaagca tcaggttctc agcgatggct tcgcgggccg
     4141 actggttgag gtgctggccg atggcgtgga cacgtcgcac aagggcgggc tgtggatctt
     4201 cgccagcgag ataaaccacg cgcccggtgt gcgccggggt tagtcccaca aggtcgccgc
     4261 ctgccacact gcaagcgatt gacatggccg cttcaagagc aaaaaagctc ttgccagttg
     4321 cgccaggtgc gacaagtgcg cccacggttc cggccaagaa tccgggccag atgaaatcaa
     4381 gttctggcgg ctcattggtg aaggccgcca taatgtctaa agccatgctt gtctcctttt
     4441 cagacagttg tggttaggcg ctgccggggc cgcaatcccc tgccgcgccg ctcaattttc
     4501 gcccctcgat gccagccacg ccgaagcctc ggccaaggcg tcctcatagg cgtagctggc
     4561 cgcttcttct gcgttctctg ctgtggcgta ctcggacaac gggaacactc cgccgccgac
     4621 ttcctcgccg ctttccaaca ggcgcaaccg ccagccgccg ccaaggtcgg aggggcgcgg
     4681 gtcgatctgg tagctgtatt cgatcattgg cggtaaacct ccttacgatt gccgatgcgc
     4741 ataacgagga tgcgcaaagc accgtcctcg atgctggcga tgattcgcca atccccaacg
     4801 cggtatttcc agaaggctcc cagtttggag cctttgaggg cttcgccaat gctgcgcggg
     4861 tcgtcgagct gggcgacacg gccatgcaaa aaagcgagga tgcggcgggc ggtctgctgg
     4921 tcgatcttgc ctagctcgcg ctcggcggct gggtcgagtt caaccttcca tgccatagcg
     4981 tttcatcact tcttcgagtg gcacggtttg ggttttgcct gcgcgaatgt cgatcaggcg
     5041 ttgctctgca aggtacaaat cttcgaggtc atccaagtgc tcaaggatgg cttcgcgggc
     5101 atagaaagtc ttggtccgcc ctgtggcctg cgccagtgct tcaaggcggg tttccacttc
     5161 ggcgggcagt cggattgcaa gcatgtttcg gctcctgttc tgaaaacgct atacaagtat
     5221 atcacaacca ccacgcaacg ctagagactc attcccggcg agttgtcgcg ctcgatcttc
     5281 tgcgcctttg cctgctcctg ctgtcgtaca ggctcgcgcc cttccagctt ctcccactgc
     5341 tgccggtact tgccaagatc ggcggcttgc cggtcgcctg cggcgctgta ggcatagcgg
     5401 gcggtgcgct gcgcgtagtc gttccaatcg cggccctcgt ctttctgccg ggtggccggt
     5461 gcgcactggc ggatagcgcc ctcgatggcc gcttgatcgt ggccggtgac gcgcatacgc
     5521 acggcaatca tggaatccac gcgggacaag tccacctcgc cgccacgctg ccgcttgagc
     5581 acgtcgcggt aatgccgctg gtaggcgtcg atcgcgctgc cgctggccgc tgcaagctcc
     5641 aaggcgggct tggctttgct gcgctcgggc tgctgcgcct tcaaggccgc ttgccgctga
     5701 tactcggcgt cgatctggct ggacaaggcc agcgccttga cgcactcccg gcgctcggcc
     5761 ttgagcaagc gcacctctgg atagctgccg tcctcccgct ggtgcttggg cttgcggttc
     5821 tcgtagccgg gtgcgcggtg cgggtggatg gctcccgata gcttcgggtc gccgtattca
     5881 cggttcaggg catcgctcag gcggttgccc acgtccttat cgtgggccgt ccccagcttc
     5941 gacacggtga tgatggcctg atagttgccg gggctggatt ccagcacggc ggcgggctgg
     6001 tagccgtctt tgataagccg ctccagcttc tcccggttca tgtcgtcgat gaggatgtga
     6061 tgcttcttgt ccgatagcgg cgtgtagtag aggttttcgc cccggcgctg taggcgctgc
     6121 atctccggcg tgcgctgctc gatctcctgc ggtgtgaaac cccgcgtgat gccgtccttc
     6181 ttgtcgagga tgaaggtttg cttccttccg tctgcctgca tcttgatgga cgttacccgg
     6241 tagcgctccg cgcctaccgc ttccgcgtac cgctcgaact gctttagctg ctccgatttc
     6301 atcgcctgcg ccctctcctg ctgtatccgc tgccgctcct gctggatgct ctcctgaagc
     6361 tctgcatttg tgatcttgaa gccgtgctct gctgctaact tcccgcacat ggccttgtat
     6421 tcgtcgttcc cggttacggt gaagctgccc cacttctgcg ccgatagctg caaggcggca
     6481 agcgtgctgt cccggttgcg ccagtcgtga atatcgatgc ttttgccctt gtccacgaaa
     6541 gacacacccc cgccccgccc cccttcttcc ttccggctgt aatgcacctg ctggcccacg
     6601 atttcaggct gataggcgcg aatgtcgcgc ggcgtcggcg gctcgctgcg gtcgccctcg
     6661 atgcgctgcg gctcgctggc ccggtgtcgc cactgctcgg caagctccgg gcttttctgc
     6721 atccgctgcc actgctccag gtcgggatag ggtctgaact gctgccggtg ctgctcgcgc
     6781 tgcttctggt gcttctcctt caaggcggct ttctctgcgg cctgctcggc ggcaatcacg
     6841 ctgcgcatgg cgttgagcac ttcgcccttg cctttccagt tgccgcgcat gagttcatcg
     6901 cggcgggcct tctgctgctc ggcaagttgc ttgcgctcct gctcctgccg cttgtcctgt
     6961 accagcttgt ccgcgttctt ctccgagtag tgcgccttgc gcccggtgat gtagtccttc
     7021 catcccggca cgtctggctt gatcggctcc ggctcgcgct gtgctacctg ctgacgctgc
     7081 ggcggctggt aagctccgag tcgcttttgc aacttggata ggctcgcgtc ccggtctgcg
     7141 ctgctggcct tcacgccaac atcaccgaca aagagcgtcg ccccgctgcc tgttttctcg
     7201 tagcgcatcc ccttggctgc taactcgcgg tgcaactgct cccacgtctg cgcctttttg
     7261 atgatcggtg cgccgtcctc aatggcgatg cgttcggcgg atttctcccc ggtgcggttt
     7321 tccatgtcgc gcttcggctg ggctggctgg cgcggcttgt ccttgtcgat gtgctcccgg
     7381 cctaactcac cattctccaa cacctgataa cgcccgttct gctcacgctg ccagccttgt
     7441 gcgttctcga ttcgtgctat cgccttgtgc gctgcctcaa tgtcgaagcc gttattgatc
     7501 ttcaccacct tgagcgtttc aggatgcacg cggttgatgg caaggtgcag gtgcaagttg
     7561 tccgtgtcgg cgtgcaaacc atagatggct tgatggtctt taacacctag ctcatccatg
     7621 aaaatgctta ccgcttcttc gacctgctcc gggctgggct gctcgccttc tcgccaactc
     7681 aggacatagt gattgatcgt atccttgctt cgcactgcct cttgtgacaa ggcaatcatt
     7741 tctgccgtct ggctctgtgg ttcatcggtg atgaatccac gcgccccggc atggatgcac
     7801 ttctcctgcg agttctcacg ctcaggctca cggatgtagc cagttaggcg ggaaacgcga
     7861 accgccttgc ttgctgcctt ctgcgggttc ttaaccttct tgacgatcac ggctcagttt
     7921 cccgatatag gctttcaaga ccacaagcgc cccggctgtg tccttgctgt acacgcctcc
     7981 gctctcgttg tggatgtgct taagcaatcc accgatgcgg cgcaactctt tgagcattac
     8041 cgcgtcggcg ctggcaatga tcggcctgcc aaagtagcgg cggcggacaa gctccgacat
     8101 gctcaacccg gcaaggtcgg catcctcttt caagcgggcc ttctccgctg cggtcagccg
     8161 tacattgata acggcgtcca gcggttccgg cccctgcgtt tcaaatggca tcgtgtctcc
     8221 ttctcgcggg gggcgggcgg ggtgtgtctt ggacagtaag aagcgagagc gccgcactgt
     8281 ccggggtcaa ggggataccc cttgcaggat aagccagacc gtgagaagcg aagcgccgaa
     8341 tgggcaggcg tccgttggaa ccacgtggcg aagccatgtg tgttacaacg gtcatcctgt
     8401 attgctcaac cgctctacta tcatatcagc agtttaggca cactagatgc actttgcaac
     8461 aagcataaaa ggcgctatgc tgtcaactcc gaaaccag
//
