LOCUS       sequence-388851-        8331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            E2-crimson, specR and pVS1 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8331)
  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 8331)
  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: pAC10
FEATURES             Location/Qualifiers
     source          1..8331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1043..1672
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2101..3174
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3240..3434
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4144..4689
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4590..4609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4806..4823
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5273..5292)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6270..6944
                     /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     6668..6687
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(7144..7935)
                     /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 gcgtgtttta gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg
       61 aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac
      121 ttgaccaacc aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag
      181 atcaccggca ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct
      241 ggcgacgttg tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg
      301 gacattgccg agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg
      361 gccgacacca ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag
      421 ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga
      481 ggcgtgaagt ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag
      541 ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc
      601 tcgaccctgt accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg
      661 cgcggtgcct tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat
      721 gaacgccaag aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca
      781 ttaccgaaga gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc
      841 acgtctcaac cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg
      901 cctggccggc cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg
      961 tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata
     1021 aacaaatacg caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc
     1081 aggcaagacg accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt
     1141 tctgttagtc gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga
     1201 tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat
     1261 cggccggcgc gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc
     1321 cgcgatcaag gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg
     1381 ggccaccgcc gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct
     1441 acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc
     1501 cgaggcgctg gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag
     1561 ctacccaggc actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc
     1621 tgcccgcgag gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga
     1681 ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc
     1741 agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac
     1801 tttcagttgc cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag
     1861 accattaccg agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga
     1921 ataaatgagt agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc
     1981 aggcaccgac gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag
     2041 cggctgggtt gtctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc
     2101 gtgagcggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg
     2161 gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc
     2221 cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg
     2281 gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc
     2341 gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt
     2401 ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac
     2461 gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg
     2521 gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga
     2581 gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga
     2641 gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg
     2701 cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag
     2761 ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac
     2821 atcgagatcg agctagttga ttggatgtac cgcgagatca cagaaggcaa gaacccggac
     2881 gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac
     2941 cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac
     3001 gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc
     3061 gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc
     3121 ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg
     3181 gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg tctctttcct
     3241 gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt
     3301 gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag
     3361 aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc
     3421 ctggcctgtg cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc
     3481 cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc
     3541 cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg
     3601 tcgccactcg accgccggcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac
     3661 cagggcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3721 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3781 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3841 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3901 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3961 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccttaa taagatgatc
     4141 ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt
     4201 gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct
     4261 tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt
     4321 caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc
     4381 tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg
     4441 ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc
     4501 gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga
     4561 acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4621 gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc
     4681 ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat
     4741 cttccaggaa atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg
     4801 agcgtagcga gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac
     4861 gcaccggtgc agcctttttt ctcctgccac atgaagcact tcactgacac cctcatcagt
     4921 gccaacatag taagccagta tacactccgc tagcgctgag gtctgcctcg tgaagaaggt
     4981 gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca
     5041 cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc
     5101 acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt
     5161 cgatttattc aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat
     5221 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     5281 gttatgagcc atattcaacg ggaaacgtct tgctcgagat ctatcgatgc atgccatggt
     5341 acccgggagc tcgaattcga agcttttgcc attctcaccg gattcagtcg tcactcatgg
     5401 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     5461 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     5521 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     5581 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     5641 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct ttgttgaata
     5701 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     5761 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc
     5821 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca
     5881 gcaacacctt cttcacgagg cagacctcag cgccagaagg ccgccagaga ggccgagcgc
     5941 ggccgtgagg cttggacgct agggcagggc atgaaaaagc ccgtagcggg ctgctacggg
     6001 cgtctgacgc ggtggaaagg gggaggggat gttgtctaca tctgaatcat gcgcggatga
     6061 cacgaactca cgacggcgat cacagacatt aacccacagt acagacactg cgacaacgtg
     6121 gcaattcgtc gcaataccgt ctcactgaac tggccgataa ttgcagacga acgggtgaaa
     6181 caaaacggtt gacaacatga agtaaacacg gtacgatgta ccacatgaaa cgacagtgag
     6241 tcaatacaga aacagaggag atattacata tggatagcac tgagaacgtc atcaagccct
     6301 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     6361 gcgtgggcga gggcaagccc tacgagggca cccagaccgc caagctgcaa gtgaccaagg
     6421 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcttctac ggctccaagg
     6481 cgtacatcaa gcaccccgcc gacatccccg actacctcaa gcagtccttc cccgagggct
     6541 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     6601 cctccctgca ggacggcacc ctcatctacc acgtgaagtt catcggcgtg aacttcccct
     6661 ccgacggccc cgtaatgcag aagaagactc tgggctggga gccctccact gagcgcaact
     6721 acccccgcga cggcgtgctg aagggcgaga accacatggc gctgaagctg aagggcggcg
     6781 gccactacct gtgtgagttc aagtccatct acatggccaa gaagcccgtg aagctgcccg
     6841 gctaccacta cgtggactac aagctcgaca tcacctccca caacgaggac tacaccgtgg
     6901 tggagcagta cgagcgcgcc gaggcccgcc accacctgtt ccagactcac ggtatggacg
     6961 aattgtacaa gcacgacgaa ttgtaaatcc gtaatcgtta atccgcaaat aacgtaaaaa
     7021 cccgcttcgg cgggtttttt tatgggggga gtttagggaa agagcatttg tcagctggat
     7081 cagcagatgg gaaccggata ctcctcagcc tccagagtag ccaatgagac gccgagcggc
     7141 cgcttatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     7201 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
     7261 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     7321 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     7381 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     7441 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     7501 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     7561 ggctggctcg aagatacccg caagaatgtc attgcgctgc cattctccaa attgcagttc
     7621 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     7681 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     7741 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     7801 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     7861 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     7921 caccgcttcc ctcataatgt ttaactttgt tttagggcga ctgccctgct gcgtaacatc
     7981 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
     8041 ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     8101 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     8161 cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg
     8221 tttccacggt gtgcgtcacc cggcaacctt gggtagcagc gaagtcgagg catttctgtc
     8281 ctggctggtc atgaccaaaa tcccttaacg tgttcctggc gttttcttgt c
//
