LOCUS       sequence-343351-        4789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MusF2 prenyltransferases from Nostoc sp. UHCC 0398.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4789)
  AUTHORS   Mattila A, Andsten RM, Jumppanen M, Assante M, Jokela J, Wahlsten M,
            Mikula KM, Sigindere C, Kwak DH, Gugger M, Koskela H, Sivonen K, Liu
            X, Yli-Kauhaluoma J, Iwai H, Fewer DP
  TITLE     Biosynthesis of the Bis-Prenylated Alkaloids Muscoride A and B.
  JOURNAL   ACS Chem Biol. 2019 Dec 20;14(12):2683-2690. doi: 
            10.1021/acschembio.9b00620. Epub 2019 Nov 19.
  PUBMED    31674754
REFERENCE   2  (bases 1 to 4789)
  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"; doi: 
            "10.1021/acschembio.9b00620"; journalName: "ACS Chem Biol"; date: 
            "2019-12-20- 20"; volume: "14"; issue: "12"; pages: "2683-2690"
COMMENT     Sequence Label: pBHRSF228
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             124..414
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             1343..1387
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1425..1443)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1439..1486
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1716..2531)
                     /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(1716..2531)
                     /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     2439..2458
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2532..2623)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2639..3388)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2649..2666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2889..2908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3584..4666)
                     /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     4628..4647
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4667..4744)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 actagtgact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct
      181 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag aaatcttctt taagatcaaa
      241 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg
      301 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat
      361 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggatccgtg
      421 acatctaatc cgattcaaac taatagcgaa cagtacttag aatacattgg taaacataaa
      481 cttgcttttg atatttacga agatttatat cctcttaagc tgtttgaaga ttttgtagaa
      541 gtagaggcta aaaaaagagg tttattttac attctttcta atgtagataa agatgaaata
      601 tatccggctc ggttctgttt gagatttcct tcactagaag aagctcaact gctttataat
      661 ccacaacaac aactacaaac cgcccttaac ttttttcgtc aagtggaaag ccgaccagaa
      721 gttaagctca attatcacca tattcagcaa ttttttggta ctatttctga ctttcaaggt
      781 attgttttga tggctgttgc catagatgcc cgtacagtaa ttactgagtc gcgtctaaag
      841 ctgtatattt ggctcaaaaa tgctccggaa aaagtagaaa cagccattgc tttgtgtggt
      901 gatagtccaa cattgcgggc atttttagtt aatgatcaac tgcaagtagg ttttgattta
      961 tttttcaatg gtgaaagcga aatagaagtt tatccaatca tcagtcaaga tgagttgcag
     1021 cagtttcata ttcgcgatcg aataatccca ctcttaccac ctcgtgcttt accgctacta
     1081 cagcaatgtg cagtttttca agtaggtttt agtgaagcga atgaaagtaa tattctctat
     1141 tttgattatg tccatgaccc caatagcttt gtcgataatc tcggtaatga gatgactaag
     1201 aaaattcatg cttattatcg ccatcagcct attaaatctt tgacagtagg cattcccgaa
     1261 cacaattttt atggacgggc aattgagcat gtgaaactat attatgatat gaattaagct
     1321 tgcggccgca ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat
     1381 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
     1441 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     1501 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     1561 gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac gaccgggtct
     1621 ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1681 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     1741 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1801 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1861 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1921 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     1981 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2041 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcgg
     2101 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2161 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2221 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2281 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2341 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2401 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2461 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     2521 atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2581 atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat gcagcgctct
     2641 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggtgtca
     2701 gctcactcaa aagcggtaat acggttatcc acagaatcag gggataaagc cggaaagaac
     2761 atgtgagcaa aaagcaaagc accggaagaa gccaacgccg caggcgtttt tccataggct
     2821 ccgcccccct gacgagcatc acaaaaatcg acgctcaagc cagaggtggc gaaacccgac
     2881 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2941 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3001 tcatagctca cgctgttggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3061 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3121 gtccaacccg gtaagacacg acttatcgcc actggcagca gccattggta actgatttag
     3181 aggactttgt cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca gattttggtg
     3241 agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag cgaaccttga
     3301 gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa tcaatcggtc
     3361 tatcaagtca acgaacagct attccgttac tctagatttc agtgcaattt atctcttcaa
     3421 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc
     3481 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     3541 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     3601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3661 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     3721 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     3781 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     3841 tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg
     3901 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     3961 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     4021 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     4081 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     4141 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     4201 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     4261 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     4321 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     4381 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     4441 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     4501 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     4561 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     4621 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     4681 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     4741 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaat
//
