LOCUS       sequence-438552-        6085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in response to 4'-O-Methylnorbelladine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6085)
  AUTHORS   d'Oelsnitz S, Diaz DJ, Kim W, Acosta DJ, Dangerfield TL, Schechter 
            MW, Minus MB, Howard JR, Do H, Loy JM, Alper HS, Zhang YJ, Ellington
            AD
  TITLE     Biosensor and machine learning-aided engineering of an 
            amaryllidaceae enzyme.
  JOURNAL   Nat Commun. 2024 Mar 7;15(1):2084. doi: 10.1038/s41467-024-46356-y.
  PUBMED    38453941
REFERENCE   2  (bases 1 to 6085)
  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 
            Commun."; date: "2024-03-7"; volume: "15(1):2084. doi"; pages: " 
            10.1038/s41467-024-46356-y"
COMMENT     Sequence Label: pSens-4OmeNB
FEATURES             Location/Qualifiers
     source          1..6085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(832..866)
                     /label=BBa_J23112
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0"
     promoter        complement(832..866)
                     /label=BBa_J23103
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     CDS             complement(1294..2109)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1294..2109)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2017..2036
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2110..2212)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2604..2621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2738..3283)
                     /direction=LEFT
                     /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     complement(2818..2837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3508..4590)
                     /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     4552..4571
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4591..4668)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4705..4724
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             5836..5853
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 cttggaccaa aacgaaaaaa ggccgctttc gcggcctctt ttctggaatt tggtaccgag
       61 ctcacagcgg cgatagtcag atagctagac cgtatgttac cgagcctgca ctcctcgaat
      121 tcttaggatt attactgctc ttcgcgcgta agtgcgcgcc acatagcctc gaagcccaac
      181 gcaatgtact caccagcgcg agccgggtcg cgcgcagcga aatccatagt cgtctcagca
      241 agcgccaaga acaacccgtc gccgaaggcg cggtactcgt cggacataaa caccataaga
      301 accccacggt ggtccaggtc gcgtaactcc gggaacatat catccgcgcg ttgttcggtt
      361 tccttcgtca acttttcaga aaccgccaac tgacgaatgg cacgatggcg agctgggtgg
      421 ttcaatcccc agctaatata actgttccag ataaaacggg tcatcatctt agcgtcagta
      481 atagaacgat ccaattccat gatcattgat tggcacatgt cctgggtcaa atgtaagtaa
      541 agggtgttga tcaactcatc tttcgttgcg aaatagcgga acaacgtccc ttccgcaact
      601 cccgcattgc gtgcaattac agcggtacta gcggcaatgc ctgattgcgc gatggcttga
      661 gttgccgctt caagcaatgc ctgctttttg tcctcagact ttgggcgagc aaccatatac
      721 taacctcctt ctgatacgtg gttccgttaa acaaaattat ttgtagaggc cccatttcgt
      781 ccttttggac tcatcagggg tggtacacac caccctatgg ggctcgtaat tgctagcata
      841 atccctagga ctgagctagc tatcagggta cttttcaaag gtcgacagcc catgcccttg
      901 cgttcggcag gtgtacaatg atacgaggta atgaagatga agtccataca atcgatagat
      961 tgggaccaaa acgaaaaaag gggagcggtt tcccgctccc ctcttttctg gaatttggta
     1021 ccgagtcgca cctgattgcc cgacattatc gcacggtgtc tcatctctga taacgcatat
     1081 tgtcgttaga actcggcgcg gccgctcaca ctgcttccgg tagtcaataa accggtaaac
     1141 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     1201 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag
     1261 gcgtttaagg gcaccaataa ctgccttaaa aaattagaaa aactcatcga gcatcaaatg
     1321 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     1381 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     1441 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     1501 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     1561 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     1621 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcggtc
     1681 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     1741 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     1801 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     1861 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     1921 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     1981 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2041 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     2101 atggctcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     2161 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     2221 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
     2281 ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc
     2341 tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt
     2401 ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg
     2461 ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     2521 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     2581 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     2641 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     2701 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     2761 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     2821 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     2881 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     2941 tccgggcagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     3001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     3061 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     3121 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     3181 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     3241 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     3301 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     3361 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
     3421 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     3481 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3541 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3601 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     3661 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     3721 tttgatggtg gttaacggcg ggatataaca tgagctatct tcggtatcgt cgtatcccac
     3781 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     3841 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     3901 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     3961 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4021 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4081 gcccagtcgc gtaccatctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4141 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4201 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4261 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     4321 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     4381 actagaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     4441 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     4501 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     4561 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     4621 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     4681 gctctccctt atgcgacgcg gccgcggcat cagagcagat tgtactgtgt cctcaatggt
     4741 tcgttgtgat ggcggtagga atgtaatcgt taatccgcaa ataacgtaaa aacccgcttc
     4801 ggcgggtttt tttatggggg gagtttaggg aaagagcatt tgtcatcccg ttgaatatgg
     4861 ctcgcatctt atcgagcata ctatcacgtc ggcgaccact agtcagttaa cgcaagggca
     4921 tgggctgtcg acctttgaaa agtaccttga cggcgtatct ttgctttcta taatgagtgc
     4981 ttactcactc atacaatagt cagtcataag tctgggctaa gcccactgat gagtcgctga
     5041 aatgcgacga aacttatgac ctctacaaat aattttgttt aacgtaaacc tccgggttaa
     5101 taaggagtaa ttatggcatc caagggcgag gagctcttta ctggcgtagt accaattctc
     5161 gtagagctcg atggcgatgt aaatggccat aagttttccg tacgcggcga gggcgagggc
     5221 gatgcaacta acggcaagct cactctcaag tttatttgta ctactggcaa gctcccagta
     5281 ccatggccaa ctctcgtaac tactctgacc tatggcgtac aatgtttttc ccgctatcca
     5341 gatcacatga agcaacatga tttttttaag tccgcaatgc cagagggcta tgtacaagag
     5401 cgcactatta gctttaagga tgatggcacc tataagactc gcgcagaggt aaagtttgag
     5461 ggcgatactc tcgtaaatcg cattgagctc aagggcattg attttaagga ggatggcaat
     5521 attctcggcc ataagctgga gtataatttc aattcccata atgtatatat taccgcagat
     5581 aagcaaaaga atggcattaa ggcgaatttt aagattcgcc ataatgtgga ggatggctcc
     5641 gtacaactcg cagatcatta tcaacaaaat actccaattg gcgatggccc agtactcctc
     5701 ccagataatc attatctctc cactcaatcc gtgctctcca aagatccaaa tgagaagcgc
     5761 gatcacatgg tactcctgga gtttgtaact gcagcaggca ttactcatgg catggatgag
     5821 ctctataagc tcgagcacca ccaccaccac cactgataat ccaaacctgt tatatgttag
     5881 ctgagactag ttggaagtgt ggctgtcctc aagcgtttta gttcgtcggt cagtttcacc
     5941 tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag aaagatgaat
     6001 gactgtcggc cattcgatgg tgtcgggtag cataacccct tgtgatacct ttgccatgtt
     6061 tcagaaacaa ctctggcgca tcggg
//
