LOCUS       sequence-457688-        6892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2,4-Diacetylphloroglucinol (DAPG)-regulatable expression of 
            SARS-CoV-2 RBD omicron variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6892)
  AUTHORS   Tominaga M, Shima Y, Nozaki K, Ito Y, Someda M, Shoya Y, Hashii N, 
            Obata C, Matsumoto-Kitano M, Suematsu K, Matsukawa T, Hosoya K, 
            Hashiba N, Kondo A, Ishii J
  TITLE     Designing strong inducible synthetic promoters in yeasts.
  JOURNAL   Nat Commun. 2024 Dec 19;15(1):10653. doi: 
            10.1038/s41467-024-54865-z.
  PUBMED    39702268
REFERENCE   2  (bases 1 to 6892)
  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: "2024-12-19"; volume: "15(1):10653. doi"; pages: " 
            10.1038/s41467-024-54865-z"
COMMENT     Sequence Label: pMT1219
FEATURES             Location/Qualifiers
     source          1..6892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3646..3702,3703..3900)
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDAAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKR"
     CDS             4528..4545
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4623..4869
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             5266..6288
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(6498..6515)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6498..6514)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6507..6529)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6723..6742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6842..6864
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctttgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccgctagc tcatcaagag agctgaacag gaaatagacg tcttgatgcc
     2281 cggttatact cacttgcaaa gagctcaacc aatcagatgg tctcactggt tgagcatgta
     2341 tgctacctat ttcactgaag attatgagag actgaatcaa atcgttaaaa ggttgaacaa
     2401 atccccattg ggagctggag ctttggctgg tcatccttat ggaattgatc gtgaatacat
     2461 tgctgagaga ttagggtttg attctgttat tggtaattct ttggccgctg tttcagacag
     2521 agattttgta gtcgaaacca tgttctggtc ttcgttgttt atgaatcata tttctcgatt
     2581 ctcagaagat ttgatcattt actccactgg agagtttgga tttatcaagt tggcagatgc
     2641 ttattctact ggatcttctc tgatgcctca aaaaaaaaac ccagactctt tggagttatt
     2701 gaggggtaaa tctggtagat gttttggggc cttggctggt ttcctcatgt ctattaagtc
     2761 cattccgtca acctataaca aagatatgca agaggataag gagcctttat ttgatactct
     2821 aatcactgta gagcactcga ttttgatagc atccggtgta gtttctacct tgaacattga
     2881 tgccgaacga atgaagaatg ctctaactat ggatatgctg gctacagatc ttgccgacta
     2941 tttagttaga aggggagttc cattcagaga aactcaccac atttctggtg aatgtgtcag
     3001 acaagccgag gagttgaacc tttctggtat tgatcagttg tccctcgaac aattgaaatc
     3061 cattgactcc cgttttgagg ctgatgtggc ttcaacgttt gactttgaag ccagtgttga
     3121 aaaaagaact gccaccggag gaacttctaa gactgctgtt ttaaagcaat tggatgcact
     3181 gaatgaaaag ctagagtctt gactgcaggt cgactctaga ggatccatga tacgaaacgt
     3241 accgtatcgt taaggtgaat gatacgaaac gtaccgtatc gttaaggtca atgatacgaa
     3301 acgtaccgta tcgttaaggt caatgatacg aaacgtaccg tatcgttaag gtgtatgata
     3361 cgaaacgtac cgtatcgtta aggtagatga tacgaaacgt accgtatcgt taaggtggcg
     3421 ttgtgatccg gatgataatg cgcaaaattt aactgttcta acccctactt gacagcaata
     3481 tataaacaga aggaagctgc cctgtcttaa accttttttt ttatcatcat tattagctta
     3541 ctttcataat tgcgactggt tccaattgac aagcttttga ttttaacgac ttttaacgac
     3601 aacttgagaa gatcaaaaaa caactaatta ttcgaaacga ctagtatgag atttccttca
     3661 atttttactg cagttttatt cgcagcatcc tccgcattag ctgctccagt caacactaca
     3721 acagaagatg aaacggcaca aattccggct gaagctgtca tcggttactc agatttagaa
     3781 ggggatttcg atgctgctgt tttgccattt tccaacagca caaataacgg gttattgttt
     3841 ataaatacta ctattgccag cattgctgct aaagaagaag gggtatcttt ggataaaaga
     3901 atcaccaact tgtgcccatt cgacgaggtt ttcaacgcta ctagattcgc ttccgtttac
     3961 gcctggaaca gaaagagaat ctccaactgc gttgctgact actccgtctt gtataacctg
     4021 gctccattct tcaccttcaa gtgctacggt gtttccccaa ctaagttgaa cgacctgtgt
     4081 ttcactaacg tctacgccga ctccttcgtt attagaggtg acgaggttag acagatcgct
     4141 ccaggtcaaa ctggtaatat cgctgactac aactacaagc tgccagacga cttcaccggt
     4201 tgtgttattg cttggaactc caacaagctg gactccaagg tttccggtaa ctacaattac
     4261 ctgtaccgtc tgttcagaaa gtccaacctg aagccattcg agagagacat ctccaccgaa
     4321 atctaccaag ctggtaacaa gccatgtaac ggtgttgccg gtttcaactg ttacttccca
     4381 ttgagatcct acagcttccg tccaacttac ggtgttggtc atcagccata cagagtcgtc
     4441 gttttgtcct tcgagttgtt gcatgctcca gctactgttt gcggtccaaa gaagtccact
     4501 aacctggtca agaacaaggg tggtggtcat catcatcacc atcactagac gcgtccttag
     4561 acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt gctagattct
     4621 aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt tttgatactt
     4681 ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct tctcgtacga
     4741 gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa
     4801 aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag tacagaagat
     4861 taagtgagac gttcgtttgt gcaagcttag atccccgggt accgagctcg aattcgccgt
     4921 accacttcaa aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg
     4981 tcgttaatta cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt
     5041 cttcgtcgaa aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt
     5101 tttgattttt ttctctttcg atgacctccc attgatattt aagttaataa acggtcttca
     5161 atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta cttcttgctc
     5221 attagaaaga aagcatagca atctaatcta agttttaatt acaaaatgaa gaagccagag
     5281 ttgactgcta cttccgttga gaagttcttg atcgagaagt tcgactccgt ttccgacttg
     5341 atgcaattgt ctgaaggtga agagtccaga gctttctcct ttgacgttgg tggtagaggt
     5401 tacgttttga gagttaattc ctgtgctgac ggtttctaca aggacagata cgtttacaga
     5461 cacttcgctt ccgctgcttt gccaattcca gaggttttgg acattggtga gttctccgag
     5521 tccttgactt actgtatctc cagaagagct cagggagtta cattgcaaga cttgccagaa
     5581 actgagttgc cagctgtttt gcaaccagtt gctgaagcta tggacgctat tgctgctgct
     5641 gacttgtctc aaacttctgg ttttggtcca ttcggtccac agggtattgg tcagtacact
     5701 acttggagag acttcatctg tgctatcgct gacccacatg tttaccactg gcaaactgtt
     5761 atggacgata ctgtttccgc ttccgttgct caggctttgg acgaattgat gttgtgggct
     5821 gaagattgtc ctgaggttag acatttggtt cacgctgact tcggttccaa caacgttttg
     5881 actgacaacg gtagaatcac tgctgttatc gactggtccg aagctatgtt cggtgactct
     5941 caatacgagg ttgctaacat cttcttttgg agaccatggt tggcttgtat ggaacagcag
     6001 actagatact tcgagagaag acacccagaa ttggctggtt ccccaagatt gagagcttac
     6061 atgttgagaa tcggtttgga ccagttgtac cagtccttgg ttgatggtaa cttcgatgat
     6121 gctgcttggg ctcaaggtag atgtgacgct atcgttagat ccggtgctgg tactgttggt
     6181 agaactcaga ttgctagaag atccgctgct gtttggactg acggttgtgt tgaagttttg
     6241 gctgactccg gtaacagaag accatccact agaccaagag ctaaagaggt tggtagagtt
     6301 taagtattaa gaacgttatt tatatttcaa atttttcttt tttttctcta cagacgctgt
     6361 acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaagatcc
     6421 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6481 ccgcttcctc ggaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     6541 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     6601 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     6661 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     6721 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6781 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6841 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc ga
//
