LOCUS       sequence-388740-        8193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            GFP, ampR and pVS1 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8193)
  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 8193)
  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: pAC13
FEATURES             Location/Qualifiers
     source          1..8193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1062..1691
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2120..3193
                     /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      3259..3453
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4163..4708
                     /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     4609..4628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4825..4842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5292..5311)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(join(7178..7969,7970..8038))
                     /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     7801..7820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8039..8143)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt gcgactagaa
       61 ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct atgcccgcgt
      121 cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac
      181 caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc tggccaggat
      241 gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc gcctggcccg
      301 cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg cgggcctgcg
      361 tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt
      421 gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg
      481 cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca ccccggcaca
      541 gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag aggcggctgc
      601 actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac
      661 gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc
      721 cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca ccaggacggc
      781 caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc ggccgggtac
      841 gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct ggccggtttg
      901 tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac cgagcgccgc
      961 cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat
     1021 atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac
     1081 ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc
     1141 aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt
     1201 gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg
     1261 accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg
     1321 cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc
     1381 caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag cagcgcattg
     1441 aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc
     1501 gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt cttgagtccc
     1561 gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc gttcttgaat
     1621 cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa
     1681 aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc
     1741 cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc
     1801 agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc tgaagatgta
     1861 cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg cgcagctacc
     1921 agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa ggaggcggca
     1981 tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg
     2041 gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc actggaaccc
     2101 ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc
     2161 ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg
     2221 catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa
     2281 agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga
     2341 cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag
     2401 catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat
     2461 ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag
     2521 tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg
     2581 ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt
     2641 actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg
     2701 cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg
     2761 cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga
     2821 aaccgggcgg ccggagtaca tcgagatcga gctagttgat tggatgtacc gcgagatcac
     2881 agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg
     2941 catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg
     3001 gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt
     3061 caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc
     3121 ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc
     3181 cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg
     3241 tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg
     3301 gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta
     3361 agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat
     3421 taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga
     3481 gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg
     3541 gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg
     3601 gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc tgaggtctgc ctcgtgaaga
     3661 aggtgttgct gactcatacc agggcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3721 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3781 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3841 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3901 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3961 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4021 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4081 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4141 accccttaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
     4201 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
     4261 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
     4321 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
     4381 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
     4441 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
     4501 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
     4561 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg
     4621 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg
     4681 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg
     4741 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct
     4801 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg
     4861 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt
     4921 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgagg
     4981 tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc catcatccag
     5041 ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc agttggtgat
     5101 tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc
     5161 cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg
     5221 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     5281 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgagatc
     5341 tatcgatgca tgccatggta cccgggagct cgaattcgaa gcttttgcca ttctcaccgg
     5401 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     5461 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     5521 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     5581 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     5641 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     5701 acggcggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct
     5761 tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac
     5821 ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat
     5881 tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctcagc gccagaaggc
     5941 cgccagagag gccgagcgcg gccgtgaggc ttggacgcta gggcagggca tgaaaaagcc
     6001 cgtagcgggc tgctacgggc gtctgacgcg gtggaaaggg ggaggggatg ttgtctacat
     6061 ctgaatcatg cgcggatgac acgaactcac gacggcgatc acagacatta acccacagta
     6121 cagacactgc gacaacgtgg caattcgtcg caatacaacg ctttggcagt ttattcttga
     6181 catgtagtga gggggctggt ataatcacat agtactgtta tacagaaaca gaggagatat
     6241 tacatatgag taaaggagaa gagcttttca caggagttgt cccaatcctc gtggaattag
     6301 acggtgatgt taatgggcac aagttctctg tcagtggaga gggtgaaggc gacgcaacat
     6361 atggcaagct gacccttaaa tttatttgca ccacgggtaa actacctgtt ccatggccaa
     6421 cactggtcac tacgttcggg tatggggttc agtgctttgc gcgctaccca gatcacatga
     6481 aacagcacga ctttttcaag agtgcaatgc ccgaaggcta tgtacaggag agaaccatct
     6541 tttttaagga tgacggcaac tataagacac gcgccgaagt gaagttcgag ggtgataccc
     6601 ttgttaatag aatcgagtta aagggtattg actttaagga agatggaaat attttaggcc
     6661 acaaactgga atataactat aactcccata atgtgtacat tatggccgac aagcaaaaga
     6721 acggtatcaa ggttaacttc aagatcagac acaacattga ggatggaagc gttcaactag
     6781 ccgaccatta ccaacaaaac accccaattg gcgatgggcc tgtgctgtta ccagacaacc
     6841 attacctgtc cactcaatct gccctttcga aagatcccaa cgaaaagcgc gaccacatgg
     6901 tccttcttga gtttgtcacg gctgctggga ttacacacgg catggatgaa ctatacaaat
     6961 aaatccgtaa tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg
     7021 gggggagttt agggaaagag catttgtcag ctggaaatct gctcgtcagt ggcgagcggc
     7081 cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7141 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7201 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7261 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg
     7321 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7381 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7441 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7501 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7561 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7621 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7681 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7741 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7801 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7861 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7921 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7981 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8041 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8101 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8161 tgccacctgt catgaccaaa atcccttaac gtg
//
