LOCUS       sequence-252690-        8231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cuminic acid-inducible system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8231)
  AUTHORS   Gonzalez LM, Mukhitov N, Voigt CA
  TITLE     Resilient living materials built by printing bacterial spores.
  JOURNAL   Nat Chem Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi: 
            10.1038/s41589-019-0412-5.
  PUBMED    31792444
REFERENCE   2  (bases 1 to 8231)
  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: "Nat Chem 
            Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi: 
            10.1038/s41589-019-0412-5."
COMMENT     Sequence Label: pLG371
FEATURES             Location/Qualifiers
     source          1..8231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1246..1332
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1351..1378
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    1479..1506
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    1540..1567
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     misc_RNA        1574..1623
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             1671..2387
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKY"
     primer_bind     complement(1698..1726)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2321..2341
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2437..2531
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4158..4176)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4244..4348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4349..4417,4418..5209)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4567..4586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5379..5967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5868..5887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tactgcctga acgagaagct atcaccgccc agcctaaacg gattccccct atgcaagggt
       61 ttattgtttt ctaaaatctg attaccaatt agaatgaata tttcccaaat attaaataat
      121 aaaacaaaaa aattgaaaaa agtgtttcca ccattttttc aattttttta taattttttt
      181 aatctgttat ttaaatagtt tatagttaaa tttacatttt cattagtcca ttcaatattc
      241 tctccaagat aactacgaac tgctaacaaa attctctccc tatgttctaa tggagaagat
      301 tcagccactg catttcccgc aatatctttt ggtatgattt tacccgtgtc catagttaaa
      361 atcatacggc ataaagttaa tatagagttg gtttcatcat cctgataatt atctattaat
      421 tcctctgacg aatccataat ggctcttctc acatcagaaa atggaatatc aggtagtaat
      481 tcctctaagt cataatttcc gtatattctt ttattttttc gttttgcttg gtaaagcatt
      541 atggttaaat ctgaatttaa ttccttctga ggaatgtatc cttgttcata aagctcttgt
      601 aaccattctc cataaataaa ttcttgtttg ggaggatgat tccacggtac catttcttgc
      661 tgaataataa ttgttaattc aatatatcgt aagttgcttt tatctcctat tttttttgaa
      721 ataggtctaa ttttttgtat aagtatttct ttactttgat ctgtcaatgg ttcagatacg
      781 acgactaaaa agtcaagatc actatttggt tttagtccac tctcaactcc tgatccaaac
      841 atgtaagtac caataaggtt attttttaaa tgtttccgaa gtattttttt cactttatta
      901 atttgttcgt atgtattcaa atatatcctc ctcactattt tgattagtac ctattttata
      961 tccatagttg ttaattaaat aaacttaatt tagtttattt atagatttca ttggcttcta
     1021 aattttttat ctagataata attattttag ttaattttat tctagattat atatgatatg
     1081 atctttcatt tccataaaac taaagtaagt gtaaacctat tcattgtttt aaaaatatct
     1141 cttgccagtc acgttacgtt attagttata gttattataa catgtattca cgaacgaaaa
     1201 tcgccattcg ccagctgcag gaattcgacc tctagcttga ggcatcaaat aaaacgaaag
     1261 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     1321 agtaggacaa atccgccgct ctagctaagc agaaggccat cctgacggat ggcctttttg
     1381 cgtttctaca aactcttgtt aactctagag ctgcctgccg cgtttcggtg atgaagatct
     1441 tcccgatgat taattaattc agaacgctcg gttgccgcaa caaacagaca atctggtctg
     1501 tttgtattat ttgactttat ctacaaggtg tggcataata acaaacagac aatctggtct
     1561 gtttgtatta tagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa
     1621 cagcctctac aaataatttt gtttaatcgg ctgaaaggtg gtgaactact atgaaaggag
     1681 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     1741 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     1801 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttcg
     1861 cgtatggtct tcaatgcttt gcgagatacc cagatcatat gaaacagcat gactttttca
     1921 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
     1981 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     2041 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     2101 ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact
     2161 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     2221 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat
     2281 ctaagctttc gaaagatccc aacgaaaaga gggaccacat ggtccttctt gagtttgtaa
     2341 cagctgctgg gattacacat ggcatggatg aactatacaa atactagtaa gcttgcggcc
     2401 gcgtcgtgac tgggaaaacc ctggcgacta gtcttggact cctgttgata gatccagtaa
     2461 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     2521 ttggtgagaa tccaggggtc cccaataatt acgatttaaa ttggagctta gtcgacagct
     2581 agccgcatgc aagctaattc ggtggaaacg aggtcatcat ttccttccga aaaaacggtt
     2641 gcatttaaat cttacatatg taatactttc aaagactaca tttgtaagat ttgatgtttg
     2701 tagcgcccgt cggctgaaag gtggtgaact actatgagcc cgaaacgtcg tacccaggca
     2761 gaacgtgcaa tggaaaccca gggtaaactg attgcagcag cactgggtgt tctgcgtgaa
     2821 aaaggttatg caggttttcg tattgcagat gttccgggtg cagccggtgt tagccgtggt
     2881 gcacagagcc atcattttcc gaccaaactg gaactgctgc tggcaacctt tgaatggctg
     2941 tatgagcaga ttaccgaacg tagccgtgca cgtctggcaa aactgaaacc ggaagatgat
     3001 gttattcagc agatgctgga tgatgcagca gaattttttc tggatgatga ttttagcatc
     3061 ggcctggatc tgattgttgc agcagatcgt gatccggcac tgcgtgaagg tattcagcgt
     3121 accgttgaac gtaatcgttt tgttgttgaa gatatgtggc tgggtgtgct ggtgagccgt
     3181 ggtctgagcc gtgatgatgc cgaagatatt ctgtggctga tttttaacag cgttcgtggt
     3241 ctggtagttc gtagcctgtg gcagaaagat aaagaacgtt ttgaacgtgt gcgtaatagc
     3301 accctggaaa ttgcacgtga acgttatgca aaattcaaac gttgataagg atcctaattg
     3361 gtaacgaatc agacaattga cggcgcgcaa cgcaattaat gtgagttagg atcctgagcg
     3421 ccggtcgcta ccattaccag ttggtctggt gtcaaaaata ataataaccg ggcaggccat
     3481 gtctgcccgt atttcgcgta aggcgatcag accagttttt aatttgtgtg tttccatgtg
     3541 tccagtttgg aatactctta acctcattgg aaatcgcggc ataatcactg gtggtatgat
     3601 tgatgaccgc gtcaacaatg acctttatgc catattcttc agcggctgca cacatttctt
     3661 taaattcttg ttcagtacct aagtaacggt tgccaatttg atacgatgtc ggctgataca
     3721 gccagtacca gttcgacatg cttttatctc cttgattccc ttcctttact tggttaatcg
     3781 gagatgtctg aatggctgta tatcctgcat catgaatatc cttcatattg tgttttaacg
     3841 tattgaacga ccaattccat gcatgaagaa tggttccgct tttgatcgac ggtgctgtaa
     3901 gctcattcga tttgttcgcc gtttcagcac tcgcagccgc cggtcctgcc agaaccaaat
     3961 gaaacagcaa taaaaatcca gcgaataacg gcagtaaaga ggttttgaat cgttttgcaa
     4021 acattcttga cactccttat ttgatttttt gaagacttac ttcggagtca aaaatccctc
     4081 ttacttcatt cttccgcttc ctcctttcaa accgatgtga agactggaga attttgttaa
     4141 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4201 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4261 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4321 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4381 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4441 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4501 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4561 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4621 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4681 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4741 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4801 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4861 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     4921 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4981 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5041 taaatctgga gccggtgagc gtgggtcgcg cggtatcatt gcagcactgg ggccagatgg
     5101 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5161 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5221 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5281 ggtgaagatc cttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5341 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5401 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5461 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5521 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5581 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5641 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5701 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5761 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5821 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5881 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5941 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggcccgacc tcgagctgga
     6001 tacttcccgt ccgccagggg gacatgccgg cgatgctgaa ggtcgcgcgc attcccgatg
     6061 aagaggccgg ttaccgcctg tttgaggata tagtaatctt tctaaatagc tttggattgg
     6121 aggagtatgg ccactaatac taagttcagc taataaaaaa atttgctaaa gaactccagc
     6181 tggatttcac tgatgagaat atcgtcggag ataaatataa taattccacg gactatagac
     6241 tatactagta tactccgtct actgtacgat acacttccgc tcaggtcctt gtcctttaac
     6301 gaggattgtt acgtacgcta atggcgtcaa aacaaagact ctagacctag gccttaagat
     6361 ctgatcatat gcatccgcgg gcccgggtta acgcgtaatc catggatcaa gagacaggat
     6421 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gtgccctgaa
     6481 tgaactgcag aaagagctgg tagttggcgc actgttcgaa gaactgccga tgtccagtaa
     6541 gattcttact atgctggttg aaccggatgc tggtaaagct acttgggttg ctgcttctac
     6601 ttatggtacc gatacaacta ctggtgagga agttaaagga gctcttaaag aaatccactt
     6661 cagtaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     6721 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggt ataaatcgcc
     6781 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     6841 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     6901 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     6961 actaagaaaa tgccgtcaaa tccgctcgcc atgacttcac taacgatgcc tttgaaaatc
     7021 ttcaagttct tttctactaa ttcaaggcgt gtctcaccag gtttttggtt tgctccggcg
     7081 caaatgcaga caatatcagc atccttgcag ggtatgtttc tctttgatgt ctttttgttt
     7141 gtgaagtatt tcacatttat attgtgcaac acttcacaaa cttttgcaag agaaaagttt
     7201 tgtctgattt atgaacaaaa aagaaaccat cattgatggt ttctttcggt aagtcccgtc
     7261 tagccttgcc ctcaatgggg aagagaaccg cttaagcccg agtcattata taaaccattt
     7321 agcacgtaat caaagccagg ctgattctga ccgggcactt gggcgctgcc attattaaaa
     7381 atcacttttg cgttggttgt atccgtgtcc gcaggcagcg tcagcgtgta aattccgtct
     7441 gcatttttag tcattggttt tccaggccaa gatccggtca attcaattac tcggctccca
     7501 tcatgtttat agatataagc atttcctggc tccaatgatt cggattttga tagccgatgg
     7561 ttttggccga cgctggatct cttttaacaa aactgtattt ctcggtcctc gttacaccat
     7621 cactgttcgt tccttttaac atgatggtgt atgttttgcc aaatggatct ccttttccga
     7681 ttgtgaattg atctccatcc ttaaacgcct gtcgtctggt ccattattga tttgataaac
     7741 ggcttttgtt gtattcgcat ctgcacgcaa ggtaatcgtc agttgatcat tgaaagaatg
     7801 tgttacacct gttttgtaat tctcaaggaa aacatgaggc gcttttgcaa tatcatcagg
     7861 ataaagcaca gctacagacc tggcattgat cgtgcctgtc agtttaccat cgttcacttg
     7921 aaatgaaccc gctccagctt tattgtcata cctgccatca ggcaattttg ttgccgtatt
     7981 gatagagaca gaggatgaac ctgcatttgc cagcacaacg ccatgtgagc cgcgctgatt
     8041 cataaatatc tggttgtttc cattcgggtt cgagagttcc tcaggctgtc cagccatcac
     8101 attgtgaaat ctattgaccg cagtgatagc ctgatcttca aataaagcac tcccgcgatc
     8161 gcctatttgg cttttccccg ggaacctcac accatttccg cctccctcag gtctggaaaa
     8221 gaaaagaggc g
//
