LOCUS       sequence-388726-        7872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Honey bee gut symbiont broad-host range vector with constitutive 
            GFP, ampR and RSF1010 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  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 7872)
  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: pAC08
FEATURES             Location/Qualifiers
     source          1..7872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(347..1318)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGHDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            2557..2644
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      2986..3380
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     CDS             complement(join(5013..5804,5805..5873))
                     /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     5636..5655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5874..5978)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(6031..6882)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(6869..7708)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
ORIGIN
        1 atatcgcgct tggcgtacag cgtcagggct ggccagcagg tcgccggtct gcttgtcctt
       61 ttggtctttc atatcagtca ccgagaaact tgccggggcc gaaaggcttg tcttcgcgga
      121 acaaggacaa ggtgcagccg tcaaggttaa ggctggccat atcagcgact gaaaagcggc
      181 cagcctcggc cttgtttgac gtataaccaa agccaccggg caaccaatag cccttgtcac
      241 ttttgatcag gtagaccgac cctgaagcgc ttttttcgta ttccataaaa cccccttctg
      301 tgcgtgagta ctcatagtat aacaggcgtg agtaccaacg caagcactac atgctgaaat
      361 ctggcccgcc cctgtccatg cctcgctggc ggggtgccgg tgcccgtgcc agctcggccc
      421 gcgcaagctg gacgctgggc agacccatga ccttgctgac ggtgcgctcg atgtaatccg
      481 cttcgtggcc gggcttgcgc tctgccagcg ctgggctggc ctcggccatg gccttgccga
      541 tttcctcggc actgcggccc cggctggcca gcttctgcgc ggcgataaag tcgcacttgc
      601 tgaggtcatg accgaagcgc ttgaccagcc cggccatctc gctgcggtac tcgtccagcg
      661 ccgtgcgccg gtggcggcta agctgccgct cgggcagttc gaggctggcc agcctgcggg
      721 ccttctcctg ctgccgctgg gcctgctcga tctgctggcc agcctgctgc accagcgccg
      781 ggccagcggt ggcggtcttg cccttggatt cacgcagcag cacccacggc tgataaccgg
      841 cgcgggtggt gtgcttgtcc ttgcggttgg tgaagcccgc caagcggcca tagtggcggc
      901 tgtcggcgct ggccgggtcg gcgtcgtact cgctggccag cgtccgggca atctgccccc
      961 gaagttcacc gcctgcggcg tcggccacct tgacccatgc ctgatagttc ttcgggctgg
     1021 tttccactac cagggcaggc tcccggccct cggctttcat gtcatccagg tcaaactcgc
     1081 tgaggtcgtc caccagcacc agaccatgcc gctcctgctc ggcgggcctg atatacacgt
     1141 cattgccctg ggcattcatc cgcttgagcc atggcgtgtt ctggagcact tcggcggctg
     1201 accattcccg gttcatcatc tggccggtgg tggcgtccct gacgccgata tcgaagcgct
     1261 cacagcccat ggccttgagc tgtcggccta tggcctgcaa agtcctgtcg ttcttcatcg
     1321 ggccaccaag cgcagccaga tcgagccgtc ctcggttgtc agtggcgtca ggtcgagcaa
     1381 gagcaacgat gcgatcagca gcaccaccgt aggcatcatg gaagccagca tcacggttag
     1441 ccatagcttc cagtgccacc cccgcgacgc gctccgggcg ctctgcgcgg cgctgctcac
     1501 ctcggcggct acctcccgca actctttggc cagctccacc catgccgccc ctgtctggcg
     1561 ctgggctttc agccactccg ccgcctgcgc ctcgctggcc tgctgggtct ggctcatgac
     1621 ctgccgggct tcgtcggcca gtgtcgccat gctctgggcc agcggttcga tctgctccgc
     1681 taactcgttg atgcctctgg atttcttcac tctgtcgatt gcgttcatgg tctattgcct
     1741 cccggtattc ctgtaagtcg atgatctggg cgttggcggt gtcgatgttc agggccacgt
     1801 ctgcccggtc ggtgcggatg ccccggcctt ccatctccac cacgttcggc cccaggtgaa
     1861 caccgggcag gcgctcgatg ccctgcgcct caagtgttct gtggtcaatg cgggcgtcgt
     1921 ggccagcccg ctctaatgcc cggttggcat ggtcggccca tgcctcgcgg gtctgctcaa
     1981 gccatgcctt gggcttgagc gcttcggtct tctgtgcccc gcccttctcc ggggtcttgc
     2041 cgttgtaccg cttgaaccac tgagcggcgg gccgctcgat gccgtcattg atccgctcgg
     2101 agatcatcag gtggcagtgc gggttctcgc cgccaccggc atggatggcc agcgtatacg
     2161 gcaggcgctc ggcaccggtc aggtgctggg cgaactcgga cgccagcgcc ttctgctggt
     2221 cgagggtcag ctcgaccggc agggcaaatt cgacctcctt gaacagccgc ccattggcgc
     2281 gttcatacag gtcggcagca tcccagtagt cggcgggccg ctcgacgaac tccggcatgt
     2341 gcccggattc ggcgtgcaag acttcatcca tgtcgcgggc atacttgcct tcgcgctgga
     2401 tgtagtcggc cttggccctg gccgattggc cgcccgacct gctgccggtt ttcgccgtaa
     2461 ggtgataaat cgccatgctg cctcgctgtt gcttttgctt ttcggctcca tgcaatggcc
     2521 ctcggagagc gcaccgcccg aagggtggcc gttaggccag tttctcgaag agaaaccggt
     2581 aagtgcgccc tcccctacaa agtagggtcg ggattgccgc cgctgtgcct ccatgatagc
     2641 ctacgagaca gcacattaac aatggggtgt caagatggtt aaggggagca acaaggcggc
     2701 ggatcggctg gccaagctcg aagaacaacg agcgcgaatc aatgccgaaa ttcagcgggt
     2761 gcgggcaagg gaacagcagc aagagcgcaa gaacgaaaca aggcgcaagg tgctggtggg
     2821 ggccatgatt ttggccaagg tgaacagcag cgagtggccg gaggatcggc tcatggcggc
     2881 aatggatgcg taccttgaac gcgaccacga ccgcgccttg ttcggtctgc cgccacgcca
     2941 gaaggatgag ccgggctgaa tgatcgaccg agacaggccc tgcggggctg cacacgcgcc
     3001 cccacccttc gggtaggggg aaaggccgct aaagcggcta aaagcgctcc agcgtatttc
     3061 tgcggggttt ggtgtggggt ttagcgggct ttgcccgcct ttccccctgc cgcgcagcgg
     3121 tggggcggtg tgtagcctag cgcagcgaat agaccagcta tccggcctct ggccgggcat
     3181 attgggcaag ggcagcagcg ccccacaagg gcgctgataa ccgcgcctag tggattattc
     3241 ttagataatc atggatggat ttttccaaca ccccgccagc ccccgcccct gctgggtttg
     3301 caggtttggg ggcgtgacag ttattgcagg ggttcgtgac agttattgca ggggggcgtg
     3361 acagttattg caggggttcg tgacagttag tacgggagtg acgggcactg gctggcaatg
     3421 tctagcaacg gcaggcattt cggctgaggg taaaagaact ttccgctaag cgatagactg
     3481 tatgtaaaca cagtattgca aggacgcgga acatgcctca tgtggcggcc aggacggcca
     3541 gccgggatcg ggatactggt cgttaccaga gccaccgacc cgagcaaacc cttctctatc
     3601 agatcgttga cgagtattac ccggcattcg ctgcgcttat ggcagagcag ggaaaggaat
     3661 tgccgggcta tgtgcaacgg gaatttgaag aatttctcca atgcgggcgg ctggagcatg
     3721 gctttctacg ggttcgctgc gagtcttgcc acgccgagca cctggtcgct ttcagaaatc
     3781 aatctaaagt atatatgagt aaacttggtc tgacaggccc ctgaattcgc atctagactg
     3841 atgagacgtg gtagagccac aaacagccgg tacaagcaac gatctccagg accatctgaa
     3901 tcatgcgcgg atgacacgaa ctcacgacgg cgatcacaga cattaaccca cagtacagac
     3961 actgcgacaa cgtggcaatt cgtcgcaata caacgctttg gcagtttatt cttgacatgt
     4021 agtgaggggg ctggtataat cacatagtac tgttatacag aaacagagga gatattacat
     4081 atgagtaaag gagaagagct tttcacagga gttgtcccaa tcctcgtgga attagacggt
     4141 gatgttaatg ggcacaagtt ctctgtcagt ggagagggtg aaggcgacgc aacatatggc
     4201 aagctgaccc ttaaatttat ttgcaccacg ggtaaactac ctgttccatg gccaacactg
     4261 gtcactacgt tcgggtatgg ggttcagtgc tttgcgcgct acccagatca catgaaacag
     4321 cacgactttt tcaagagtgc aatgcccgaa ggctatgtac aggagagaac catctttttt
     4381 aaggatgacg gcaactataa gacacgcgcc gaagtgaagt tcgagggtga tacccttgtt
     4441 aatagaatcg agttaaaggg tattgacttt aaggaagatg gaaatatttt aggccacaaa
     4501 ctggaatata actataactc ccataatgtg tacattatgg ccgacaagca aaagaacggt
     4561 atcaaggtta acttcaagat cagacacaac attgaggatg gaagcgttca actagccgac
     4621 cattaccaac aaaacacccc aattggcgat gggcctgtgc tgttaccaga caaccattac
     4681 ctgtccactc aatctgccct ttcgaaagat cccaacgaaa agcgcgacca catggtcctt
     4741 cttgagtttg tcacggctgc tgggattaca cacggcatgg atgaactata caaataaatc
     4801 cgtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg
     4861 agtttaggga aagagcattt gtcagctgga aatctgctcg tcagtggcga gcggccgcga
     4921 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgggaccca
     5161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6001 cctgtcatga ccaaaatccc ttaacgtgag tcagcctgcc gccttgggcc gggtgatgtc
     6061 gtacttgccc gccgcgaact cggttaccgt ccagcccagc gcgaccagct ccggcaacgc
     6121 ctcgcgcacc cgctggcggc gcttgcgcat ggtcgaacca ctggcctctg acggccagac
     6181 atagccgcac aaggtatcta tggaagcctt gccggttttg ccggggtcga tccagccaca
     6241 cagccgctgg tgcagcaggc gggcggtttc gctgtccagc gcccgcacct cgtccatgct
     6301 gatgcgcaca tgctggccgc cacccatgac ggcctgcgcg atcaaggggt tcagggccac
     6361 gtacaggcgc ccgtccgcct cgtcgctggc gtactccgac agcagccgaa acccctgccg
     6421 cttgcggcca ttctgggcga tgatggatac cttccaaagg cgctcgatgc agtcctgtat
     6481 gtgcttgagc gccccaccac tatcgacctc tgccccgatt tcctttgcca gcgcccgata
     6541 gctacctttg accaccatgg catcagcggt gacggcctcc cacttgggtt ccaggaacag
     6601 ccggagctgc cgtccgcctt cggtcttggg ttccgggcca agcactaggc cattaggccc
     6661 agccatggcc accagccctt gcaggatgcg cagatcatca gcgcccagcg gctccgggcc
     6721 gctgaactcg atccgcttgc cgtcgccgta gtcatacgtc acgtccagct tgctgcgctt
     6781 gcgctcgccc cgcttgaggg cacggaacag gccgggggcc agacagtgcg ccgggtcgtg
     6841 ccggacgtgg ctgaggctgt gcttgttctt aggcttcacc acggggcacc cccttgctct
     6901 tgcgctgcct ctccagcacg gcgggcttga gcaccccgcc gtcatgccgc ctgaaccacc
     6961 gatcagcgaa cggtgcgcca tagttggcct tgctcacacc gaagcggacg aagaaccggc
     7021 gctggtcgtc gtccacaccc cattcctcgg cctcggcgct ggtcatgctc gacaggtagg
     7081 actgccagcg gatgttatcg accagtaccg agctgccccg gctggcctgc tgctggtcgc
     7141 ctgcgcccat catggccgcg cccttgctgg catggtgcag gaacacgata gagcacccgg
     7201 tatcggcggc gatggcctcc atgcgaccga tgacctgggc catggggccg ctggcgtttt
     7261 cttcctcgat gtggaaccgg cgcagcgtgt ccagcaccat caggcggcgg ccctcggcgg
     7321 cgcgcttgag gccgtcgaac cactccgggg ccatgatgtt gggcaggctg ccgatcagcg
     7381 gctggatcag caggccgtca gccacggctt gccgttcctc ggcgctgagg tgcgccccaa
     7441 gggcgtgcag gcggtgatga atggcggtgg gcgggtcttc ggcgggcagg tagatcaccg
     7501 ggccggtggg cagttcgccc acctccagca gatccggccc gcctgcaatc tgtgcggcca
     7561 gttgcagggc cagcatggat ttaccggcac caccgggcga caccagcgcc ccgaccgtac
     7621 cggccaccat gttgggcaaa acgtagtcca gcggtggcgg cgctgctgcg aacgcctcca
     7681 gaatattgat aggcttatgg gtagccattg attgcctcct ttgcaggcag ttggtggtta
     7741 ggcgctggcg gggtcactac ccccgccctg cgccgctctg agttcttcca ggcactcgcg
     7801 cagcgcctcg tattcgtcgt cggtcagcca gaacttgcgc tgacgcatcc ctttggcctt
     7861 catgcgctcg gc
//
