LOCUS       sequence-437622-        3986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mutant Norbelladine 4'-O-Methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3986)
  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 3986)
  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: pNb4OMT-A53M
FEATURES             Location/Qualifiers
     source          1..3986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             24..215
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     372..391
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(466..483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(637..1225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(717..736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_RNA        1494..1543
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    complement(2482..2515)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        2526..2597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2598..2666,2667..3458)
                     /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(2816..2835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    complement(3592..3625)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
       61 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gatgcggatg
      121 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
      181 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg cagacggtca
      241 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      301 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
      361 gcttaactat gcggcatcag agcagattgt actgagagtg caccggtgtg aaataccgca
      421 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
      481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      661 cgagcatcac aaaaattgac gctcaagtca gaggtggcga aacccgacag gactataaag
      721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1261 tcagtggaac gaaaactcac gttaaggccc tctccaagac cgagccatca acaaagcgtc
     1321 tcgctgaggt ttcatggagc ctctggttca tctccggcaa ttaaaaaagc ggctaaccac
     1381 gccgcttttt ttacgtctgc aggaacgggc tgtcgaccgg cgtgccgttc gtttaccgct
     1441 agctcagtcc taggtacaat tacagccatc gtacgagccc tggctgagca cagctgtcac
     1501 cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac aaataatttt
     1561 gtttaaacta gtgaaccacg aggcctacat atgggtgctt caattgatga ctactccttg
     1621 gtacataaaa acatcttgca ttccgaagat ctgctgaaat acattcttga aaccagtgca
     1681 tatcctcgcg aacacgaaca attaaagggt ctgcgtgagg ttactgaaaa gcacgaatgg
     1741 tcatccatgt tagtaccggc agacgaaggt ttattcctgt caatgttact gaagttgatg
     1801 aatgcaaaac gtactatcga aatcggcgtg tacacgggat acagcctgtt aacaacggca
     1861 ttggctttac cggaggatgg taaaattacg gcgatcgatg taaataagag ttattacgaa
     1921 atcggattgc cctttattca gaaggcgggc gtggaacaca agatcaactt tattgagtcg
     1981 gaagcgcttc ccgtgctgga tcaaatgtta gaggagatga aggaggagga tttatacgac
     2041 tacgcttttg tggatgctga taaaagcaat tatgccaatt accatgagcg tcttgtaaaa
     2101 cttgtacgta tcggtggtgc catcctgtac gacaatacac tgtggtatgg ttctgttgcg
     2161 tacccggaat accccggtct gcatccagag gaagaagtcg cgcgtctgag ctttcgtaac
     2221 ttgaatacct ttttagcagc agatcctcgc gtagaaatta gtcaagtctc aattggtgat
     2281 ggcgtgacca tttgtcgccg cttgtattaa taatcctatc gccactttca gccaaaaaac
     2341 ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg gtggacagga tcggcggttt
     2401 tcttttctct tctcaacacc cttcgcgtca acacttttcc gccaaggaga cggttggtca
     2461 ggttttcggg aggtgtggct ggaagttcct atactttcta gagaatagga acttctttct
     2521 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2581 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2641 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2701 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2761 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2821 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2881 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2941 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3001 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3061 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3121 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3181 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3241 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3301 ccggtgagcg tggatcgcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3361 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3421 tcgctgagat aggtgcctca ctgattaagc attggtaagt tgtgatggcg gtaggaatgt
     3481 aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt
     3541 ttagggaaag agcatttgtc atcccgttga atatggctcc cttaacgtga ggaagttcct
     3601 atactttcta gagaatagga acttctacag atggacttgg gttggcggtt tcaggagtct
     3661 gcaaaacgct gcgacctgag caacaacatg aatggtcatc ggtttccgtg tttcgtaaag
     3721 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
     3781 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg
     3841 agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc
     3901 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat
     3961 cggtatcatt acccccatga acagaa
//
