LOCUS       sequence-388714-        8757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible two-plasmid system bearing CP25 > lacI, ampR, pTF-FC2
            replicon (2/2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8757)
  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 8757)
  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: pAC17V5b
FEATURES             Location/Qualifiers
     source          1..8757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             161..1243
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(180..199)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      1263..1311
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(1517..2308,2309..2377))
                     /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     2140..2159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2378..2482)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4758..4769)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             4976..4984
                     /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 acgacgcgct cggctttggc agtttattct
       61 tgacatgtag tgagggggct ggtataatca catagtactg ttatacagaa acagaggaga
      121 tattcggtac gcaagtacac aagatacagg agaggtactc atgaaaccag taacgttata
      181 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
      241 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
      301 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
      361 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
      421 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
      481 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
      541 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
      601 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
      661 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
      721 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
      781 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
      841 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
      901 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
      961 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     1021 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     1081 ctctcagggc caggcggtga agggcaatca gctgttgccg gtgtcactgg tgaaaagaaa
     1141 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1201 gcagctggca cgacaggttt cccgactgga aagcgggcag tgatgtaaaa gcaagctgat
     1261 aaaccgatac aattaaaggc tccttttgga gccttttttt ttggagattt tcaacatgaa
     1321 gaaattatta tttgatgatc agatagcggc ggggaactgc cagacatcaa ataaaacaaa
     1381 aggctcagtc ggaagactgg gccttttgtt tcgagcggcc gcgattatca aaaaggatct
     1441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1621 gcttaccatc tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag
     1681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1801 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2101 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     2341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     2401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2461 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgtc atgaccaaaa
     2521 tcccttaacg tgctcaaggc agccagaaca tccaaaaatc cgctgtaggc gattctgatg
     2581 gcttctgttc ggggtatgag gcgatgtgct ccacaagcac gaccgacaag ccagccgaaa
     2641 cgctcgccaa gctggccgct gcgctacctg ccggagaggc caaacgctac cgggtgatgg
     2701 tgtccattgt cccggtggaa gaatgaaggc gagcggggcg gggcggggtg tgtctggccg
     2761 ggcttgtccc ggccttgtct tttggggttt ttggtaagga cttttgacgc cctcaagggc
     2821 ggcgaaaagt accgcgccta gtggatgatc ttggatagat catggaggga ttttgcgaat
     2881 cgcccgcaaa cccgcgccgt tgctaggttt ttcagatttc ggcggggaac gttacgccgg
     2941 gggttgggga acgttacgcc gggggttggg gaacgttacg ccgggggttg ggcctgctga
     3001 tttcccactt gcccttggcg tactcgttca ccgtccaacc aacggcggca agctcgacaa
     3061 gcgccttgcg ggcggtctgg cggcgctttt tcattgcttc atcgttggct gcgtctggcc
     3121 atacataacc gcacagcgtg tcgatttcag ctttgccaga ctttccgggg tcaatccagc
     3181 cacatagccg ctggtgtagc agccgtgccg ggtctgtttc cagcttgcga acttctgcca
     3241 tgttgatgcg ggtgtgcggg cgctctccca taaccgcatc cgccagccgg ggattaagcg
     3301 caacaaataa cttgccttct tgctcgtcgc tcgcgtactc ggacagaatg cggaaccctt
     3361 gccgcttacc acctctttcg acaatcaccg ataccgccca aagccgttcg atgctttccc
     3421 ggatggtttt aaactgactt cctccatctt cggcgtagcc aagttctcgg gccaactggc
     3481 ggaagctgcc cttggctacc atcgtatctt tctctatggc gtcccaccgc atatcaagcc
     3541 ataggcggag ttgctggcct gcttctgatt ccgtttcgtg ccgtagcacg atgccgcgcc
     3601 cgttatctcc ggaaattgca gccattgcca ccagcccttg caatacgcgc aagtcatcgc
     3661 cgccaagtgg ttcaggcccc caaaaacgga ttgagtcatc tccgtagttg taggtcacat
     3721 cgagcttgag cctctttcgt tctccgcgct tgagactgcg gaaaagcccc ggcgtgaggc
     3781 aatgcgcggg gtcgtgccgc gcatgggtga ggtcatagct cattgcgctt tccccctttt
     3841 tttcctgctc cattgctgat tgcctcctgt agttcaacgg gcaacaacac cccgccgctg
     3901 tgccgctgat accagcggtc tagcggggtc gcgtcgtagt tctgcttgct cacgccaaag
     3961 cggacaaaaa ggccgcgccg ctcgttgccg atgggcgaac gatcaaagcc ccggtcactc
     4021 atgcgctcgg cttcctgctc cgtcattttg gcgacatagc cgcaccatct ggcgttgtcg
     4081 atcagggcag acgcgccccg cgctgcctgc tgctggtcgg tctggccgtc gcgggcgctg
     4141 cccttgttga cgtggtgcag gtacaggaca gccgccccgg tggtcgccgc gatgtgttcc
     4201 aacacggaaa caaggtgggc catgtcgcca ttgctgttct cgtcgaggct gtgaatccgg
     4261 ctcagggtgt ccagcacaat cagccgggcc cctgcgctgt agtcgatgac gcggcgcaag
     4321 tgcgcctcgt ccatcacgtt tagccgcttg cccatgatcg gctcaagcat caggttctca
     4381 gcgatggctt cgcgggccga ctggttgagg tgctggccga tggcgtggac acgtcgcaca
     4441 agggcgggct gtggatcttc gccagcgaga taaaccacgc gcccggtgtg cgccggggtt
     4501 agtcccacaa ggtcgccgcc tgccacactg caagcgattg acatggccgc ttcaagagca
     4561 aaaaagctct tgccagttgc gccaggtgcg acaagtgcgc ccacggttcc ggccaagaat
     4621 ccgggccaga tgaaatcaag ttctggcggc tcattggtga aggccgccat aatgtctaaa
     4681 gccatgcttg tctccttttc agacagttgt ggttaggcgc tgccggggcc gcaatcccct
     4741 gccgcgccgc tcaattttcg cccctcgatg ccagccacgc cgaagcctcg gccaaggcgt
     4801 cctcataggc gtagctggcc gcttcttctg cgttctctgc tgtggcgtac tcggacaacg
     4861 ggaacactcc gccgccgact tcctcgccgc tttccaacag gcgcaaccgc cagccgccgc
     4921 caaggtcgga ggggcgcggg tcgatctggt agctgtattc gatcattggc ggtaaacctc
     4981 cttacgattg ccgatgcgca taacgaggat gcgcaaagca ccgtcctcga tgctggcgat
     5041 gattcgccaa tccccaacgc ggtatttcca gaaggctccc agtttggagc ctttgagggc
     5101 ttcgccaatg ctgcgcgggt cgtcgagctg ggcgacacgg ccatgcaaaa aagcgaggat
     5161 gcggcgggcg gtctgctggt cgatcttgcc tagctcgcgc tcggcggctg ggtcgagttc
     5221 aaccttccat gccatagcgt ttcatcactt cttcgagtgg cacggtttgg gttttgcctg
     5281 cgcgaatgtc gatcaggcgt tgctctgcaa ggtacaaatc ttcgaggtca tccaagtgct
     5341 caaggatggc ttcgcgggca tagaaagtct tggtccgccc tgtggcctgc gccagtgctt
     5401 caaggcgggt ttccacttcg gcgggcagtc ggattgcaag catgtttcgg ctcctgttct
     5461 gaaaacgcta tacaagtata tcacaaccac cacgcaacgc tagagactca ttcccggcga
     5521 gttgtcgcgc tcgatcttct gcgcctttgc ctgctcctgc tgtcgtacag gctcgcgccc
     5581 ttccagcttc tcccactgct gccggtactt gccaagatcg gcggcttgcc ggtcgcctgc
     5641 ggcgctgtag gcatagcggg cggtgcgctg cgcgtagtcg ttccaatcgc ggccctcgtc
     5701 tttctgccgg gtggccggtg cgcactggcg gatagcgccc tcgatggccg cttgatcgtg
     5761 gccggtgacg cgcatacgca cggcaatcat ggaatccacg cgggacaagt ccacctcgcc
     5821 gccacgctgc cgcttgagca cgtcgcggta atgccgctgg taggcgtcga tcgcgctgcc
     5881 gctggccgct gcaagctcca aggcgggctt ggctttgctg cgctcgggct gctgcgcctt
     5941 caaggccgct tgccgctgat actcggcgtc gatctggctg gacaaggcca gcgccttgac
     6001 gcactcccgg cgctcggcct tgagcaagcg cacctctgga tagctgccgt cctcccgctg
     6061 gtgcttgggc ttgcggttct cgtagccggg tgcgcggtgc gggtggatgg ctcccgatag
     6121 cttcgggtcg ccgtattcac ggttcagggc atcgctcagg cggttgccca cgtccttatc
     6181 gtgggccgtc cccagcttcg acacggtgat gatggcctga tagttgccgg ggctggattc
     6241 cagcacggcg gcgggctggt agccgtcttt gataagccgc tccagcttct cccggttcat
     6301 gtcgtcgatg aggatgtgat gcttcttgtc cgatagcggc gtgtagtaga ggttttcgcc
     6361 ccggcgctgt aggcgctgca tctccggcgt gcgctgctcg atctcctgcg gtgtgaaacc
     6421 ccgcgtgatg ccgtccttct tgtcgaggat gaaggtttgc ttccttccgt ctgcctgcat
     6481 cttgatggac gttacccggt agcgctccgc gcctaccgct tccgcgtacc gctcgaactg
     6541 ctttagctgc tccgatttca tcgcctgcgc cctctcctgc tgtatccgct gccgctcctg
     6601 ctggatgctc tcctgaagct ctgcatttgt gatcttgaag ccgtgctctg ctgctaactt
     6661 cccgcacatg gccttgtatt cgtcgttccc ggttacggtg aagctgcccc acttctgcgc
     6721 cgatagctgc aaggcggcaa gcgtgctgtc ccggttgcgc cagtcgtgaa tatcgatgct
     6781 tttgcccttg tccacgaaag acacaccccc gccccgcccc ccttcttcct tccggctgta
     6841 atgcacctgc tggcccacga tttcaggctg ataggcgcga atgtcgcgcg gcgtcggcgg
     6901 ctcgctgcgg tcgccctcga tgcgctgcgg ctcgctggcc cggtgtcgcc actgctcggc
     6961 aagctccggg cttttctgca tccgctgcca ctgctccagg tcgggatagg gtctgaactg
     7021 ctgccggtgc tgctcgcgct gcttctggtg cttctccttc aaggcggctt tctctgcggc
     7081 ctgctcggcg gcaatcacgc tgcgcatggc gttgagcact tcgcccttgc ctttccagtt
     7141 gccgcgcatg agttcatcgc ggcgggcctt ctgctgctcg gcaagttgct tgcgctcctg
     7201 ctcctgccgc ttgtcctgta ccagcttgtc cgcgttcttc tccgagtagt gcgccttgcg
     7261 cccggtgatg tagtccttcc atcccggcac gtctggcttg atcggctccg gctcgcgctg
     7321 tgctacctgc tgacgctgcg gcggctggta agctccgagt cgcttttgca acttggatag
     7381 gctcgcgtcc cggtctgcgc tgctggcctt cacgccaaca tcaccgacaa agagcgtcgc
     7441 cccgctgcct gttttctcgt agcgcatccc cttggctgct aactcgcggt gcaactgctc
     7501 ccacgtctgc gcctttttga tgatcggtgc gccgtcctca atggcgatgc gttcggcgga
     7561 tttctccccg gtgcggtttt ccatgtcgcg cttcggctgg gctggctggc gcggcttgtc
     7621 cttgtcgatg tgctcccggc ctaactcacc attctccaac acctgataac gcccgttctg
     7681 ctcacgctgc cagccttgtg cgttctcgat tcgtgctatc gccttgtgcg ctgcctcaat
     7741 gtcgaagccg ttattgatct tcaccacctt gagcgtttca ggatgcacgc ggttgatggc
     7801 aaggtgcagg tgcaagttgt ccgtgtcggc gtgcaaacca tagatggctt gatggtcttt
     7861 aacacctagc tcatccatga aaatgcttac cgcttcttcg acctgctccg ggctgggctg
     7921 ctcgccttct cgccaactca ggacatagtg attgatcgta tccttgcttc gcactgcctc
     7981 ttgtgacaag gcaatcattt ctgccgtctg gctctgtggt tcatcggtga tgaatccacg
     8041 cgccccggca tggatgcact tctcctgcga gttctcacgc tcaggctcac ggatgtagcc
     8101 agttaggcgg gaaacgcgaa ccgccttgct tgctgccttc tgcgggttct taaccttctt
     8161 gacgatcacg gctcagtttc ccgatatagg ctttcaagac cacaagcgcc ccggctgtgt
     8221 ccttgctgta cacgcctccg ctctcgttgt ggatgtgctt aagcaatcca ccgatgcggc
     8281 gcaactcttt gagcattacc gcgtcggcgc tggcaatgat cggcctgcca aagtagcggc
     8341 ggcggacaag ctccgacatg ctcaacccgg caaggtcggc atcctctttc aagcgggcct
     8401 tctccgctgc ggtcagccgt acattgataa cggcgtccag cggttccggc ccctgcgttt
     8461 caaatggcat cgtgtctcct tctcgcgggg ggcgggcggg gtgtgtcttg gacagtaaga
     8521 agcgagagcg ccgcactgtc cggggtcaag gggatacccc ttgcaggata agccagaccg
     8581 tgagaagcga agcgccgaat gggcaggcgt ccgttggaac cacgtggcga agccatgtgt
     8641 gttacaacgg tcatcctgta ttgctcaacc gctctactat catatcagca gtttaggcac
     8701 actagatgca ctttgcaaca agcataaaag gcgctatgct gtcaactccg aaaccag
//
