LOCUS       sequence-211934-        6950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent biosensor for gauging the activity of the PQS quorum 
            sensing system in Pseudomonas aeruginosa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6950)
  AUTHORS   Yang L, Barken KB, Skindersoe ME, Christensen AB, Givskov M, 
            Tolker-Nielsen T
  TITLE     Effects of iron on DNA release and biofilm development by 
            Pseudomonas aeruginosa.
  JOURNAL   Microbiology. 2007 May;153(Pt 5):1318-28. doi: 
            10.1099/mic.0.2006/004911-0.
  PUBMED    17464046
REFERENCE   2  (bases 1 to 6950)
  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: 
            "Microbiology."; date: "2007-05"; volume: "153(Pt 5):1318-28. doi"; 
            pages: " 10.1099/mic.0.2006/004911-0"
COMMENT     Sequence Label: pAC37
FEATURES             Location/Qualifiers
     source          1..6950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     primer_bind     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(230..257)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     RBS             706..717
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             730..1443
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             730..1443
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(760..788)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1383..1403
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1508..1602
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1646..2305
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1712..1731)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2370..2456
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2491..2508)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2491..2507)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2500..2522)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2668..3501)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      3515..3866
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     promoter        join(3916..3921,3922..3938,3939..3944)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4133..4666
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(4758..4777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4877..4899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4937..4955)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5023..5127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5128..5196,5197..5988)
                     /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     complement(5346..5365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6159..6747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6648..6667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6901..6918
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgaca tgattacgaa ttgcggccgc gcgaattcaa aaaggccatc
      241 cgtcaggatg gccttctagg atcaatgccg tcgccccctt ggagcccagg ccgagcgcct
      301 cgaactgtga gatttgggag gcgatttgcc gagcaaagtg ggttgtcatt ggtttgccat
      361 ctcatgggtt cggacgaggc ctcgagcaag ggttgtaacg gtttttgtct ggccaatggg
      421 ctcttgcgta aaaaggctgc cgcccttctt gcttggttgc cgttctcgga tcccgcgcag
      481 cccggtgggt gtgccaaatt tctcgcggtt tggatcgcgc cgattgccgc ggcctacgaa
      541 gcccgtggtt cttctccccg aaactttttc gttcggactc cgaatatcgc gcttcgccca
      601 gcgccgctag tttcccgttc ctgacaaagc aagcgctctg gctcaggtat ctcctgatcc
      661 ggatgcatat cgctgaagag ggaacgttct gtcatgggta ccattaaaga ggagaaatta
      721 actatgagca tgcgtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
      781 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
      841 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
      901 ccaacacttg tcactacttt cggttatggt gttcaatgct ttgcgagata cccagatcat
      961 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
     1021 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
     1081 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     1141 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     1201 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     1261 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     1321 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac
     1381 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     1441 aaaaggcctg cagcaaacga cgaaaactac gctgcagcag tttaataagc ttaattagct
     1501 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     1561 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag
     1621 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     1681 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     1741 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1801 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1861 atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1921 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     1981 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     2041 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     2101 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     2161 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     2221 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     2281 cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc
     2341 tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact
     2401 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     2461 cgctaggagc ttgcggccgc ccagcttggc actggccgtc gttttacaac gtcgtgactg
     2521 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2581 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaaagg
     2641 caggccgggc cgtggtggcc acggcctcta ggccagatcc agcggcatct gggttagtcg
     2701 agcgcgggcc gcttcccatg tctcaccagg gcgagcctgt ttcgcgatct cagcatctga
     2761 aatcttcccg gccttgcgct tcgctggggc cttacccacc gccttggcgg gcttcttcgg
     2821 tccaaaactg aacaacagat gtgtgacctt gcgcccggtc tttcgctgcg cccactccac
     2881 ctgtagcggg ctgtgctcgt tgatctgcgt cacggctgga tcaagcactc gcaacttgaa
     2941 gtccttgatc gagggatacc ggccttccag ttgaaaccac tttcgcagct ggtcaatttc
     3001 tatttcgcgc tggccgatgc tgtcccattg catgagcagc tcgtaaagcc tgatcgcgtg
     3061 ggtgctgtcc atcttggcca cgtcagccaa ggcgtatttg gtgaactgtt tggtgagttc
     3121 cgtcaggtac ggcagcatgt ctttggtgaa cctgagttct acacggccct caccctcccg
     3181 gtagatgatt gtttgcaccc agccggtaat catcacactc ggtcttttcc ccttgccatt
     3241 gggctcttgg gttaaccgga cttcccgccg tttcaggcgc agggccgctt ctttgagctg
     3301 gttgtaggaa gattcgatag ggacacccgc catcgtcgct atgtcctccg ccgtcactga
     3361 atacatcact tcatcggtga caggctcgct cctcttcacc tggctaatac aggccagaac
     3421 gatccgctgt tcctgaacac tgaggcgata cgcggcctcg accagggcat tgcttttgta
     3481 aaccattggg ggtgaggcca cgttcgacat tccttgtgta taaggggaca ctgtatctgc
     3541 gtcccacaat acaacaaatc cgtcccttta caacaacaaa tccgtccctt cttaacaaca
     3601 aatccgtccc ttaatggcaa caaatccgtc cctttttaaa ctctacaggc cacggattac
     3661 gtggcctgta gacgtcctaa aaggtttaaa agggaaaagg aagaaaaggg tggaaacgca
     3721 aaaaacgcac cactacgtgg ccccgttggg gccgcatttg tgcccctgaa ggggcggggg
     3781 aggcgtctgg gcaatccccg ttttaccagt cccctatcgc cgcctgagag ggcgcaggaa
     3841 gcgagtaatc agggtatcga ggcggattca cccttggcgt ccaaccagcg gcaccagcgg
     3901 cgcctgagag gcgaattgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt
     3961 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     4021 gcggttttca tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag
     4081 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
     4141 cagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag gtggctcaag
     4201 tatgggcatc attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc
     4261 tcttgatctt ttcggtcgtg agttcggaga cgtagccacc tactcccaac atcagccgga
     4321 ctccgattac ctcgggaact tgctccgtag taagacattc atcgcgcttg ctgccttcga
     4381 ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg cccaagtttg agcagccgcg
     4441 tagtgagatc tatatctatg atctcgcagt ctccggcgag caccggaggc agggcattgc
     4501 caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta
     4561 cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg
     4621 ggaagaagtg atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc
     4681 cgagatcggc ttcccggccg cggagttgtt cggtaaattg tcacaacgcc gcggccaatt
     4741 cgatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4801 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4861 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4921 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4981 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5041 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5101 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5161 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5221 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5281 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5341 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5401 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5461 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5521 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5581 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5641 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5701 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5761 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5821 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5881 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5941 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6001 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6061 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6121 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6181 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6241 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6301 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6361 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6421 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6481 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6541 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6601 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6661 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6721 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6781 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6841 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6901 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
//
