LOCUS       sequence-310187-        5952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 episomal expression plasmid for S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5952)
  AUTHORS   Pullmann P, Knorrscheidt A, Munch J, Palme PR, Hoehenwarter W, 
            Marillonnet S, Alcalde M, Westermann B, Weissenborn MJ
  TITLE     A modular two yeast species secretion system for the production and 
            preparative application of unspecific peroxygenases.
  JOURNAL   Commun Biol. 2021 May 12;4(1):562. doi: 10.1038/s42003-021-02076-3.
  PUBMED    33980981
REFERENCE   2  (bases 1 to 5952)
  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: "Commun 
            Biol."; date: "2021-05-12"; volume: "4(1):562. doi"; pages: " 
            10.1038/s42003-021-02076-3"
COMMENT     Sequence Label: pAGT572_Nemo
FEATURES             Location/Qualifiers
     source          1..5952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..483
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    42..159
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     454..477
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    558..579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(594..599,600..617,618..624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    632..648
                     /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     637..659
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     656..672
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     656..672
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             668..991
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    685..741
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(742..759)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(742..758)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(751..773)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(967..986)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      2149..3026
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3035..3139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3140..3208,3209..4000)
                     /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(3358..3377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4212..4800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4701..4720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4923..5726)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5727..5947)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 tggtcactct gtgaagacaa tgccgaattc ggatccggag acggattaga agccgccgag
       61 cgggtgacag ccctccgaag ggagactctc ctccgtgcgt cctcgtcgtc accggtcgcg
      121 ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata
      181 ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa
      241 atgaacgaat caaattaaca accataggat gataatgcga ttagtttttt agccttattt
      301 ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat ataaatgcaa
      361 aaactgcata accactttaa ctaatacttt caacattttc ggtttgtatt acttcttatt
      421 caaatgtaat aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag
      481 gagaaaaaac caatgtgaga ccgcagctgg cacgacaggt ttcccgactg gaaagcgggc
      541 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
      601 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
      661 aacagctatg accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc
      721 ccgggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
      781 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
      841 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
      901 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
      961 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1021 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1081 ggtctcagct ttaaagtaag agcgctacat tggtctacct ttttgttctt ttacttaaac
     1141 attagttagt tcgttttctt tttctcattt ttttatgttt cccccccaaa gttctgattt
     1201 tataatattt tatttcacac aattccattt aacagagggg gaatagattc tttagcttag
     1261 aaaattagtg atcaatatat atttgccttt cttttcatct tttcagtgat attaatggtt
     1321 tcgagacact gcaatggccc tagttgtcta agaggataga tgttactgtc aaagatgata
     1381 ttttgaattt caattgacgt aattaatgat actattaata atacagagcg tatatgaagt
     1441 attgcaaata acatgcacag ttcttttggg atgagaatga taatgaaagg cgaaggcggg
     1501 cgttcagaaa agcgttgcgg agtaacgctg caattgtctt ctgcacgaag tggccgttat
     1561 cgatgataag ctgtcaaaca tgagaattaa ttccacggac tatagactat acctagtata
     1621 ctccgtctac tgtacgatac acttccgctc aggtccttgt cctttaacga ggccttacca
     1681 ctcttttgtt actctattga tccagctcag caaaggcagt gtgatctaag attctatctt
     1741 cgcgatgtag taaaactagc tagaccgaga aagagactag aaatgcaaaa ggcacttcta
     1801 caatggctgc catcattatt atgcgatgtg acgctgcagc ttctcaatga tattcgaata
     1861 cgctttgagg agatacagcc taatatccga caaactgttt tacagattta cgatcgtact
     1921 tgttacccat cattgaattt tgaacatccg aacctgggag ttttccctga aacagatagt
     1981 atatttgaac ctgtataata atatatagtc tagcgcttta cggaagagaa tgtatgtatt
     2041 tcggttcctg gagaaactat tgcatctatt gcataggtaa tcttgcacgt cgcatccccg
     2101 gttcattttc tgcgtttcca tcttgcactt caatagcata tctttgttaa cgaagcatct
     2161 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2221 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     2281 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2341 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     2401 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     2461 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     2521 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     2581 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     2641 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     2701 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     2761 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     2821 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     2881 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     2941 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     3001 atgtttgtgg aagcggtatt cgcaatgccg agtgcgcgga acccctattt gtttattttt
     3061 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3301 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3361 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3841 agccggtgag cgtggttctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3961 gatcgctgag ataggtgcct cactgattaa gcattggtaa tttcgcgtat ccaacggctg
     4021 cccaggcagc cgtttttatt cctgtcagac caagtttact catatatact ttagattgat
     4081 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4141 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4201 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4261 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4321 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4381 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4441 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4501 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4561 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4621 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4681 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4741 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4801 aacgccagca acgcggcctt tttacggttc ctgccttttt caatgggtaa taactgatat
     4861 aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt
     4921 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt
     4981 tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg
     5041 tcagatcctg tagataccac atcatccacg gttctatact gttgacccaa tgcgtcaccc
     5101 ttgtcatcta aacccactcc gggtgtcata atcaaccaat cgtaaccttc atctcttcca
     5161 cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt ggcaatgtca
     5221 acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac
     5281 atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca
     5341 atacctgggc ccacgacacc gtgagcattc gtaatgtctg cccattctgc tattctgtat
     5401 acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcctcg
     5461 aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg
     5521 gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct
     5581 tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt
     5641 tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca
     5701 cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg
     5761 tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata
     5821 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
     5881 tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
     5941 aattgaaaag ct
//
