LOCUS       sequence-405746-        8475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control plasmid expressing mScarlet-I fluorescent protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8475)
  AUTHORS   Geistodt-Kiener A, Totozafy JC, Le Goff G, Vergne J, Sakai K, 
            Ouazzani J, Mouille G, Viaud M, O'Connell RJ, Dallery JF
  TITLE     Yeast-based heterologous production of the colletochlorin family of 
            fungal secondary metabolites.
  JOURNAL   Metab Eng. 2023 Oct 18:S1096-7176(23)00142-8. doi: 
            10.1016/j.ymben.2023.10.002.
  PUBMED    37863177
REFERENCE   2  (bases 1 to 8475)
  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: "Metab Eng. 
            2023 Oct 18:S1096-7176(23)00142-8. doi: 
            10.1016/j.ymben.2023.10.002."
COMMENT     Sequence Label: pHYX143
FEATURES             Location/Qualifiers
     source          1..8475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(359..379)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(419..475)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(494..514)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(518..1213)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(793..812)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1062..1080
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1217..1273)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(1316..1327)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     1605..1624
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(1964..1984)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2528..2544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2528..2544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2556..2573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2727..3315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2807..2826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3563..4378)
                     /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="MSHIQRETSCSRPHLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4286..4305
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4370..4389
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        complement(4672..4776)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4803..6145
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5766..5813)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     6186..6204
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6242..6264)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6382..6787
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6788..7882
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     8139..8156
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8140..8156
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 actcgagcag gaccatgtgg tcgcgcttct cgttagggtc cttggagagg gcggactggt
       61 agctgaggta gtggttgtca gggaggagga cagggccgtc gccgataggg gtgttctgct
      121 ggtagtggtc ggcgagctgg acgccgccgt cctcgatgtt gtggcgaatc ttgaagttgg
      181 ccttgatgcc gttcttctgc ttgtcagagc cgcctccgcc tgagcctcct ccgcccccgg
      241 gttgggccag cttcttctcc agggcctggt tcttccactc cagctgggcc agcttcttct
      301 ccagggcctg cagcttcttc tccagctgct cgccggagcc gccggcggtg cccagctcca
      361 ccttgcgctt cttcttgggg ggctcgcgga tcagctcggc cttgtccata tttaaatgag
      421 ggccagggtt ttcttcaacg tcgccagcct gtttgaggag agagaagttg gtggcgccag
      481 agcctcccat cgcaccttga aaataaagat tttcgatctt gtacagctcg tccatgccgc
      541 cggtggagtg gcggccctcg gagcgttcgt actgttccac cacggtgtag tcctcgttgt
      601 gggaggtgat gtccaacttg cggtcgacgt tgtaggcgcc gggcatctgc acgggcttct
      661 tggccttgta ggtggtcttg aagtccgcca ggtagcgtcc gccgtccttc aggcgcaggg
      721 ccatcttaat gtcgcccttc agcacgccgt cctcggggta caaccgctcg gtggacgctt
      781 cccagcccat tgtcttcttc tgcattacgg ggccgtcagg agggaagttg gtgccgcgga
      841 gcttcacctt gtagatcagg gtgccgtcct ccagggaggt gtcctgggtc acggtcacgg
      901 cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggactgct
      961 tatagtagtc ggggatgtcg gcggggtgct tgatgaaggc cctggagccg tacatgaact
     1021 gaggggacag gatgtcccag gagaagggca gggggccacc cttggtcacc ttcagcttgg
     1081 cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc
     1141 cgttcatgga gccctccatg tgcaccttga accgcatgaa ctccttgatc actgcctcgc
     1201 ccttgctcac catatcaggg ccagggtttt cttcaacgtc gccagcctgt ttgaggagag
     1261 agaagttggt ggcgccagag ccgtttaaac attgagcgag ctccttcttc agggcctgca
     1321 gctcccactt cagctgcgcc agctccttct tgttggcctg cagctccttc ttcagggccc
     1381 cgggtcctcc gccagatccg ccgcctcctc tctcgatgtt gtggcgaatc ttgaagttgg
     1441 ccttgatgcc gttcttctgc ttgtcggcgg tgatgtagac gttgtgggag ttgtagttgt
     1501 actcgagctt gtggccgagg atgttgccat cttccttgaa gtcgatgccc ttcagctcga
     1561 tgcggttgac gagggtgtct ccctcgaact tgacctcagc gcgggtcttg tagttgccgt
     1621 cgtccttgaa gaaaatggtg cgctcttgga cgtagccctc aggcatggcg gacttgaaga
     1681 agtcgtgctg cttcatgtgg tcagggtagc gggcgaagca ctggaggccg tagccgaggg
     1741 tggtgacgag ggtaggccaa gggacaggga gcttgccggt ggtgcagatg agcttgaggg
     1801 tgagcttgcc gtaggtggcg tcgccctcgc cctcgccgct gacggagaac ttgtggccgt
     1861 tgacgtcgcc gtcaagctcg acgaggatag ggacgacgcc ggtgaagagt tcctcgccct
     1921 tgctgaccat cccggggccg gagccgccgg cggtacccaa ttcgaccttt ctcttctttt
     1981 ttggaggctc gggaattaat tccgctttat ccatgttgtt attttattat ggaataatta
     2041 gttgcgtgag tttggttttt ttttttttat ttttgagtta aacaaatata attattgttt
     2101 aagctgaaat aaagatatat atgtatatct gggaaaggct gttggttatc tgtcttactt
     2161 ctgtatattt aagaagtaag tagaaatact gattacacat ggaagagaag ttacacacat
     2221 tacaaagact tctctgccct tatatacctt ttttttgtgg tcttctatac tctgcccaat
     2281 gttgctatcc ccagccaagc cttacttaat acacaaacgt accattgtcc aaccagaagc
     2341 gtctaagctt tacaatcctg tttgggccct ttatccgcct atccataatg ttaagcagcc
     2401 taaaacttgc tagtctagcc ggcatctttc ccttgtataa cttaaaatac ccggagcttc
     2461 agtgatatgt agaacacagc cgcgccggat gaaaggaagc tcggtttttt cctatgcccg
     2521 ggccatggtc atagctgttt cctgtgtgac gcttcctcgc tcactgactc gctgcgctcg
     2581 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2641 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3361 gaaaactcac gttaagggat tttggtcatg acctttttac ggttcctggc cttttgctgg
     3421 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3481 gcctttgagt gagctgatac cgctcgccgc gtcaagtcag cgtaatgctc tgccagtgtt
     3541 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     3601 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     3661 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     3721 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     3781 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     3841 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     3901 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     3961 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     4021 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     4081 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     4141 gcataaattc cgtcagccag tttaaacgga ccatctcatc tgtaacatca ttggcaacgc
     4201 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     4261 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     4321 ccatgttgga atttaaatgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     4381 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     4441 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc
     4501 gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacccactc gtgcacccaa
     4561 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4621 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4681 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4741 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4801 tgaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt
     4861 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat
     4921 tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct
     4981 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc
     5041 gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga
     5101 gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa
     5161 tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt
     5221 ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac
     5281 tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg
     5341 atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc
     5401 agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta
     5461 cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta
     5521 aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa
     5581 ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga
     5641 gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag
     5701 tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc
     5761 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac ttcaaagcgt
     5821 ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc
     5881 acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag aacggcatag
     5941 tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt tatgtaggat gaaaggtagt
     6001 ctagtacctc ctgtgatatt atcccattcc atgcggggta tcgtatgctt ccttcagcac
     6061 taccctttag ctgttctata tgctgccact cctcaattgg attagtctca tccttcaatg
     6121 ctatcatttc ctttgatatt ggatcatact aagaaaccat tattatcatg acattaacct
     6181 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6241 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6301 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6361 atgcggcagg ggatcctcta gaactgtggg aatactcagg tatcgtaaga tgcaagagtt
     6421 cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta
     6481 tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg
     6541 catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagc gccggaaccg
     6601 gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca atacttgaag
     6661 ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca atcgtcttac
     6721 tttctaactt ttcttacctt ttacatttca gcaatatata tatatatttc aaggatatac
     6781 cattctaatg tctgccccta agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga
     6841 aatcacagcc gaagccatta aggttcttaa agctatttct gatgttcgtt ccaatgtcaa
     6901 gttcgatttc gaaaatcatt taattggtgg tgctgctatc gatgctacag gtgttccact
     6961 tccagatgag gcgctggaag cctccaagaa ggctgatgcc gttttgttag gtgctgtggg
     7021 tggtcctaaa tggggtaccg gtagtgttag acctgaacaa ggtttactaa aaatccgtaa
     7081 agaacttcaa ttgtacgcca acttaagacc atgtaacttt gcatccgact ctcttttaga
     7141 cttatctcca atcaagccac aatttgctaa aggtactgac ttcgttgttg tcagagaatt
     7201 agtgggaggt atttactttg gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga
     7261 tagtgaacaa tacaccgttc cagaagtgca aagaatcaca agaatggccg ctttcatggc
     7321 cctacaacat gagccaccat tgcctatttg gtccttggat aaagctaatg ttttggcctc
     7381 ttcaagatta tggagaaaaa ctgtggagga aaccatcaag aacgaattcc ctacattgaa
     7441 ggttcaacat caattgattg attctgccgc catgatccta gttaagaacc caacccacct
     7501 aaatggtatt ataatcacca gcaacatgtt tggtgatatt atctccgatg aagcctccgt
     7561 tatcccaggt tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa
     7621 caccgcattt ggtttgtacg aaccatgcca cggttctgct ccagatttgc caaagaataa
     7681 ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt
     7741 gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag gttttggatg caggtatcag
     7801 aactggtgat ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga
     7861 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt
     7921 tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg
     7981 tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatgt
     8041 aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg
     8101 cactttaaca ttaatattga caaggaggag ggcaccgttg taaaacgacg gccagtggcc
     8161 cgggcctcta gaggcatttt caacatcgta ttttccgaag cgttgcataa tgaaaatgtg
     8221 ataataaggc agtgagctac tttgtcacaa caaagtacga gaacagatga ggttccgtgg
     8281 tgtttttggg gcaaccgcac tatttggagc gctgtatata tttatatcct actacaaaaa
     8341 atcataaatg acactgaatt ggacacatgc tagcgtcaaa taccttgcct ggtaaagttg
     8401 tgtgctagtg tctcccgtct tctgtctact tgtacagctc gtccatgccg agggtgatgc
     8461 cagcggcggt gacga
//
