LOCUS       sequence-198914-        9169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Methanol inducing HSA, and estrogen inducing hGH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9169)
  AUTHORS   Cao J, Perez-Pinera P, Lowenhaupt K, Wu MR, Purcell O, de la 
            Fuente-Nunez C, Lu TK
  TITLE     Versatile and on-demand biologics co-production in yeast.
  JOURNAL   Nat Commun. 2018 Jan 8;9(1):77. doi: 10.1038/s41467-017-02587-w.
  PUBMED    29311542
REFERENCE   2  (bases 1 to 9169)
  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: "2018-01-8"; volume: "9(1):77. doi"; pages: " 
            10.1038/s41467-017-02587-w"
COMMENT     Sequence Label: JC135
FEATURES             Location/Qualifiers
     source          1..9169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        157..568
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        576..623
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             642..1016
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      1082..1329
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1098..1116)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        1344..2282
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             join(2283..2339,2340..2537)
                     /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
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKR"
     terminator      4317..4563
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     rep_origin      complement(4624..5212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4704..4723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5332..5381
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     promoter        5502..5908
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5938..5958
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7177..7326
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             7333..7359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      7401..7647
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             join(8213..8269,8270..8467)
                     /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
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKR"
     terminator      join(9065..9169,1..142)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
ORIGIN
        1 tcttctcgta cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt
       61 aggggtttgg gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca
      121 gagtacagaa gattaagtga gaccttcgtt tgtgcggatc ccccacacac catagcttca
      181 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
      241 cacttcaaaa cacccaagca cagcatacta aattttccct ctttcttcct ctagggtgtc
      301 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
      361 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaat tttttttttt
      421 agtttttttc tctttcagtg acctccattg atatttaagt taataaacgg tcttcaattt
      481 ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc ttgttcatta
      541 gaaagaaagc atagcaatct aatctaaggg gcggtgttga caattaatca tcggcatagt
      601 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
      661 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
      721 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
      781 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
      841 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
      901 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
      961 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     1021 tccgacggcg gcccacgggt cccaggcctc ggagatccgt cccccttttc ctttgtcgat
     1081 atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
     1141 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
     1201 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
     1261 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
     1321 ttaatttgca agctggagac caagatctaa catccaaaga cgaaaggttg aatgaaacct
     1381 ttttgccatc cgacatccac aggtccattc tcacacataa gtgccaaacg caacaggagg
     1441 ggatacacta gcagcagacc gttgcaaacg caggacctcc actcctcttc tcctcaacac
     1501 ccacttttgc catcgaaaaa ccagcccagt tattgggctt gattggagct cgctcattcc
     1561 aattccttct attaggctac taacaccatg actttattag cctgtctatc ctggcccccc
     1621 tggcgaggtt catgtttgtt tatttccgaa tgcaacaagc tccgcattac acccgaacat
     1681 cactccagat gagggctttc tgagtgtggg gtcaaatagt ttcatgttcc ccaaatggcc
     1741 caaaactgac agtttaaacg ctgtcttgga acctaatatg acaaaagcgt gatctcatcc
     1801 aagatgaact aagtttggtt cgttgaaatg ctaacggcca gttggtcaaa aagaaacttc
     1861 caaaagtcgg cataccgttt gtcttgtttg gtattgattg acgaatgctc aaaaataatc
     1921 tcattaatgc ttagcgcagt ctctctatcg cttctgaacc ccggtgcacc tgtgccgaaa
     1981 cgcaaatggg gaaacacccg ctttttggat gattatgcat tgtctccaca ttgtatgctt
     2041 ccaagattct ggtgggaata ctgctgatag cctaacgttc atgatcaaaa tttaactgtt
     2101 ctaaccccta cttgacagca atatataaac agaaggaagc tgccctgtct taaacctttt
     2161 tttttatcat cattattagc ttactttcat aattgcgact ggttccaatt gacaagcttt
     2221 tgattttaac gacttttaac gacaacttga gaagatcaaa aaacaactaa ttattcgaaa
     2281 cgatgagatt tccttcaatt tttactgctg ttttattcgc agcatcctcc gcattagctg
     2341 ctccagtcaa cactacaaca gaagatgaaa cggcacaaat tccggctgaa gctgtcatcg
     2401 gttactcaga tttagaaggg gatttcgatg ttgctgtttt gccattttcc aacagcacaa
     2461 ataacgggtt attgtttata aatactacta ttgccagcat tgctgctaaa gaagaagggg
     2521 tatctctcga gaagagagat gctcacaagt ccgaggttgc tcacagattc aaggacttgg
     2581 gtgaagaaaa cttcaaggct ttggttttga tcgctttcgc tcagtacttg cagcagtgtc
     2641 cattcgagga tcacgttaag ttggttaatg aggttactga gttcgctaag acttgtgttg
     2701 ctgacgaatc cgctgagaac tgtgataagt ccttgcacac tttgttcggt gacaagttgt
     2761 gtacagttgc tactttgaga gaaacttacg gtgagatggc tgactgttgt gctaagcaag
     2821 agccagaaag aaacgagtgt ttcttgcagc acaaggatga caacccaaac ttgcctagat
     2881 tggttagacc agaggttgac gttatgtgta ctgctttcca cgacaacgaa gagactttct
     2941 tgaagaagta cttgtacgag atcgctagaa gacacccata cttctacgct ccagagttgt
     3001 tgttttttgc taagagatac aaggctgctt tcactgagtg ttgtcaggct gctgacaaag
     3061 ctgcttgttt gttgccaaag ttggacgaat tgagagatga gggtaaggct tcctccgcta
     3121 agcagagatt gaagtgtgct tccttgcaga agtttggaga aagagctttt aaggcttggg
     3181 ctgttgctag attgtcccag agatttccta aggctgaatt cgctgaggtt tccaaattgg
     3241 ttactgactt gactaaggtt cacacagaat gttgtcacgg tgacttgttg gaatgtgctg
     3301 atgacagagc tgatttggct aagtacattt gtgagaacca ggactccatc tcctccaagt
     3361 tgaaagaatg ttgtgagaag cctttgttgg agaagtccca ctgtattgct gaggttgaga
     3421 acgacgaaat gccagctgac ttgccatctt tggctgctga tttcgttgag tccaaggatg
     3481 tttgtaagaa ctacgctgag gctaaggacg ttttcttggg tatgttcttg tatgaatatg
     3541 ctagaagaca tccagactac tccgttgttt tgttgttgag attggctaag acttacgaga
     3601 ctactttgga gaagtgttgt gctgctgctg acccacatga atgttacgct aaggttttcg
     3661 acgagttcaa gccattggtt gaggaaccac agaacttgat taagcagaat tgtgagttgt
     3721 tcgagcagtt gggtgagtac aagttccaga acgctttgtt ggttagatac actaagaagg
     3781 ttccacaggt ttccactcct actttggttg aagtttccag aaacttgggt aaggttggtt
     3841 ccaagtgttg taagcaccct gaagctaaga gaatgccatg tgctgaggac tacttgtctg
     3901 ttgttttgaa ccagttgtgt gttttgcacg aaaagactcc agtttccgac agagttacaa
     3961 aatgttgtac tgaatccttg gttaacagaa gaccttgttt ctccgctttg gaggttgacg
     4021 agacatacgt tccaaaagag ttcaacgctg agactttcac tttccacgct gacatctgta
     4081 ctttgtccga gaaagagaga cagattaaga agcagactgc tttggttgag ttggttaagc
     4141 acaagccaaa ggctacaaaa gagcagttga aggctgttat ggacgacttc gctgcttttg
     4201 ttgagaaatg ttgtaaggct gacgacaaag agacttgttt cgctgaagag ggtaagaagt
     4261 tggttgctgc ttctcaagct gctttgggtt tgtaaaccgg tcttgctaga ttctaatcaa
     4321 gaggatgtca gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat
     4381 ttgtaaccta tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc
     4441 tcctgatcag cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat
     4501 tcgagtttga tgtttttctt ggtatttccc actcctcttc agagtacaga agattaagtg
     4561 agacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4801 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5041 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5281 ggtcatgaga tcagatctga gctcgtttgg ccgtggccgt gctcgtcctc gtcggccggc
     5341 ttgtcgacga cggcggtctc cgtcgtcagg atcatccggg ccacaagctt gctgacagaa
     5401 gcctcaagaa aaaaaaaatt cttcttcgac tatgctggag gcagagatga tcgagccggt
     5461 agttaactat atatagctaa attggttcca tcacctcgag tccacacacc atagcttcaa
     5521 aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc
     5581 acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg
     5641 ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt
     5701 cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttg
     5761 atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt
     5821 ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta
     5881 gaaagaaagc atagcaatct aatctaagga cggcaactag taaaaatgcg cggagctcct
     5941 aagaaaaagc gcaaagtccg gccggcatct agacccgggg agcgcccctt ccagtgtcgc
     6001 atttgcatgc ggaacttttc gcgccaggac aggcttgaca ggcatacccg tactcatacc
     6061 ggtgaaaaac cgtttcagtg tcggatctgt atgcgaaatt tctcccagaa ggagcacttg
     6121 gcggggcatc tacgtacgca caccggcgag aagccattcc aatgccgaat atgcatgcgc
     6181 aacttcagtc gccgcgacaa cctgaaccgg cacctaaaaa cccacctgag gaacatatgc
     6241 ggcggaggca cacctgcagc tgcgtcgact ctagaggatc catctgctgg agacatgaga
     6301 gctgccaacc tttggccaag cccgctcatg atcaaacgct ctaagaagaa cagcctggcc
     6361 ttgtccctga cggccgacca gatggtcagt gccttgttgg atgctgagcc ccccatactc
     6421 tattccgagt atgatcctac cagacccttc agtgaagctt cgatgatggg cttactgacc
     6481 aacctggcag acagggagct ggttcacatg atcaactggg cgaagagggt gccaggcttt
     6541 gtggatttga ccctccatga tcaggtccac cttctagaat gtgcctggct agagatcctg
     6601 atgattggtc tcgtctggcg ctccatggag cacccagtga agctactgtt tgctcctaac
     6661 ttgctcttgg acaggaacca gggaaaatgt gtagagggca tggtggagat cttcgacatg
     6721 ctgctggcta catcatctcg gttccgcatg atgaatctgc agggagagga gtttgtgtgc
     6781 ctcaaatcta ttattttgct taattctgga gtgtacacat ttctgtccag caccctgaag
     6841 tctctggaag agaaggacca tatccaccga gtcctggaca agatcacaga cactttgatc
     6901 cacctgatgg ccaaggcagg cctgaccctg cagcagcagc accagcggct ggcccagctc
     6961 ctcctcatcc tctcccacat caggcacatg agtaacaaag gcatggagca tctgtacagc
     7021 atgaagtgca agaacgtggt gcccctctat gacctgctgc tggagatgct ggacgcccac
     7081 cgcctacatg cgcccactag ccgtggaggg gcatccgtgg aggagacgga ccaaagccac
     7141 ttggccactg cgggctctac ttcatcgcct agggccgacg cgctggacga tttcgatctc
     7201 gacatgctgg gttctgatgc cctcgatgac tttgacctgg atatgttggg aagcgacgca
     7261 ttggatgact ttgatctgga catgctcggc tccgatgctc tggacgattt cgatctcgat
     7321 atgttaatta actacccgta cgacgttccg gactacgctt cttgaggtac catcggtaga
     7381 ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag
     7441 gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt
     7501 ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt
     7561 gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct
     7621 cttcagagta cagaagatta agtgagagct agcatatcat atagaagtca tcgcggcaga
     7681 tcaattcatc aatcggagtg aggatattgc gagtttacca ccatcaatcg gagtgaggat
     7741 ctccaattgg tgacggtcca gtcatcaatc ggagtgagga ttcagctgct tctcgaggcc
     7801 gcacatcaat cggagtgagg atgtacaggg tgggctgttc caccatcaat cggagtgagg
     7861 atttgcgtca atggggcgga gttcatcaat cggagtgagg atatcgaagt catcgagagc
     7921 actcatcaat cggagtgagg atatactcca cccattgacg tcacatcaat cggagtgagg
     7981 attggaacca gaagggtctc ttcatcaatc ggagtgagga taccctatgg gcgcgccgtc
     8041 gctggcaata atagcgggcg gacgcatgtc atgagattat tggaaaccac cagaatcgaa
     8101 tataaaaggc gaacaccttt cccaattttg gtttcccctg acccaaagac tttaaattta
     8161 atttatttgt ccctatttca atcaattgaa caactatcaa aacacagaat tcatgagatt
     8221 tccttcaatt tttactgctg ttttattcgc agcatcctcc gcattagctg ctccagtcaa
     8281 cactacaaca gaagatgaaa cggcacaaat tccggctgaa gctgtcatcg gttactcaga
     8341 tttagaaggg gatttcgatg ttgctgtttt gccattttcc aacagcacaa ataacgggtt
     8401 attgtttata aatactacta ttgccagcat tgctgctaaa gaagaagggg tatctctcga
     8461 gaagagattc ccaaccattc ccttatctag actttttgac aacgctatgc tccgcgccca
     8521 tcgtctgcac cagctggcct ttgacaccta ccaggagttt gaagaagcct atatcccaaa
     8581 ggaacagaag tattcattcc tgcagaaccc ccagacctcc ctctgtttct cagagtctat
     8641 tccgacaccc tccaacaggg aggaaacaca acagaaatcc aacctagagc tgctccgcat
     8701 ctccctgctg ctcatccagt cgtggctgga gcccgtgcag ttcctcagga gtgtcttcgc
     8761 caacagcctg gtgtacggcg cctctgacag caacgtctat gacctcctaa aggacctaga
     8821 ggaaggcatc caaacgctga tggggaggct ggaagatggc agcccccgga ctgggcagat
     8881 cttcaagcag acctacagca agttcgacac aaactcacac aacgatgacg cactactcaa
     8941 gaactacggg ctgctctact gcttcaggaa ggacatggac aaggtcgaga cattcctgcg
     9001 catcgtgcag tgccgctctg tggagggcag ctgtggcttc tagaccggtc ttgctagatt
     9061 ctaatcaaga ggatgtcaga atgccatttg cctgagagat gcaggcttca tttttgatac
     9121 ttttttattt gtaacctata tagtatagga ttttttttgt cattttgtt
//
