LOCUS       sequence-203180-        6026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Estrogen inducing hGH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6026)
  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 6026)
  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: PP364
FEATURES             Location/Qualifiers
     source          1..6026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..410
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             434..454
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1673..1822
                     /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             1829..1855
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      1897..2143
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             join(2709..2765,2766..2963)
                     /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"
     CDS             3557..3586
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3602..3619
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3643..3889
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3904..4317
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        4324..4371
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4390..4764
                     /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      4830..5077
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4846..4864)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(5152..5740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5232..5251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5860..5909
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
ORIGIN
        1 agtccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc
       61 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc
      121 tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga
      181 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt
      241 tttcttgaaa attttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa
      301 gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac
      361 tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag gactagtaaa
      421 aatgcgcgga gctcctaaga aaaagcgcaa agtccggccg gcatctagac ccggggagcg
      481 ccccttccag tgtcgcattt gcatgcggaa cttttcgcgc caggacaggc ttgacaggca
      541 tacccgtact cataccggtg aaaaaccgtt tcagtgtcgg atctgtatgc gaaatttctc
      601 ccagaaggag cacttggcgg ggcatctacg tacgcacacc ggcgagaagc cattccaatg
      661 ccgaatatgc atgcgcaact tcagtcgccg cgacaacctg aaccggcacc taaaaaccca
      721 cctgaggaac atatgcggcg gaggcacacc tgcagctgcg tcgactctag aggatccatc
      781 tgctggagac atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa
      841 gaagaacagc ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc
      901 tgagcccccc atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat
      961 gatgggctta ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa
     1021 gagggtgcca ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc
     1081 ctggctagag atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagtgaagct
     1141 actgtttgct cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt
     1201 ggagatcttc gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg
     1261 agaggagttt gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct
     1321 gtccagcacc ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat
     1381 cacagacact ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca
     1441 gcggctggcc cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat
     1501 ggagcatctg tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga
     1561 gatgctggac gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga
     1621 gacggaccaa agccacttgg ccactgcggg ctctacttca tcgcctaggg ccgacgcgct
     1681 ggacgatttc gatctcgaca tgctgggttc tgatgccctc gatgactttg acctggatat
     1741 gttgggaagc gacgcattgg atgactttga tctggacatg ctcggctccg atgctctgga
     1801 cgatttcgat ctcgatatgt taattaacta cccgtacgac gttccggact acgcttcttg
     1861 aggtaccatc ggtagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt
     1921 tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag
     1981 gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc
     2041 agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt
     2101 ggtatttccc actcctcttc agagtacaga agattaagtg agagctagca tatcatatag
     2161 aagtcatcgc ggcagatcaa ttcatcaatc ggagtgagga tattgcgagt ttaccaccat
     2221 caatcggagt gaggatctcc aattggtgac ggtccagtca tcaatcggag tgaggattca
     2281 gctgcttctc gaggccgcac atcaatcgga gtgaggatgt acagggtggg ctgttccacc
     2341 atcaatcgga gtgaggattt gcgtcaatgg ggcggagttc atcaatcgga gtgaggatat
     2401 cgaagtcatc gagagcactc atcaatcgga gtgaggatat actccaccca ttgacgtcac
     2461 atcaatcgga gtgaggattg gaaccagaag ggtctcttca tcaatcggag tgaggatacc
     2521 ctatgggcgc gccgtcgctg gcaataatag cgggcggacg catgtcatga gattattgga
     2581 aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt cccctgaccc
     2641 aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac tatcaaaaca
     2701 cagaattcat gagatttcct tcaattttta ctgctgtttt attcgcagca tcctccgcat
     2761 tagctgctcc agtcaacact acaacagaag atgaaacggc acaaattccg gctgaagctg
     2821 tcatcggtta ctcagattta gaaggggatt tcgatgttgc tgttttgcca ttttccaaca
     2881 gcacaaataa cgggttattg tttataaata ctactattgc cagcattgct gctaaagaag
     2941 aaggggtatc tctcgagaag agattcccaa ccattccctt atctagactt tttgacaacg
     3001 ctatgctccg cgcccatcgt ctgcaccagc tggcctttga cacctaccag gagtttgaag
     3061 aagcctatat cccaaaggaa cagaagtatt cattcctgca gaacccccag acctccctct
     3121 gtttctcaga gtctattccg acaccctcca acagggagga aacacaacag aaatccaacc
     3181 tagagctgct ccgcatctcc ctgctgctca tccagtcgtg gctggagccc gtgcagttcc
     3241 tcaggagtgt cttcgccaac agcctggtgt acggcgcctc tgacagcaac gtctatgacc
     3301 tcctaaagga cctagaggaa ggcatccaaa cgctgatggg gaggctggaa gatggcagcc
     3361 cccggactgg gcagatcttc aagcagacct acagcaagtt cgacacaaac tcacacaacg
     3421 atgacgcact actcaagaac tacgggctgc tctactgctt caggaaggac atggacaagg
     3481 tcgagacatt cctgcgcatc gtgcagtgcc gctctgtgga gggcagctgt ggcttctagc
     3541 ggccgccagc tttctagaac aaaaactcat ctcagaagag gatctgaata gcgccgtcga
     3601 ccatcatcat catcatcatt gaccggtctt gctagattct aatcaagagg atgtcagaat
     3661 gccatttgcc tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata
     3721 gtataggatt ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta
     3781 tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt
     3841 tttcttggta tttcccactc ctcttcagag tacagaagat taagtgagac cttcgtttgt
     3901 gcggatcccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
     3961 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
     4021 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
     4081 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
     4141 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
     4201 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
     4261 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagggc
     4321 ggtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     4381 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     4441 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     4501 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     4561 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     4621 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     4681 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     4741 ttcgtggccg aggagcagga ctgacacgtc cgacgcggcc cgacgggtcc gaggcctcgg
     4801 agatccgtcc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     4861 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     4921 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     4981 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     5041 tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag ctggagacca acatgtgagc
     5101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5161 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5221 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5281 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5341 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5401 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5461 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5521 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5581 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5641 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5701 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5761 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatc
     5821 agatctgagc tcgtttggcc gtggccgtgc tcgtcctcgt cggccggctt gtcgacgacg
     5881 gcggtctccg tcgtcaggat catccgggcc acaagcttgc tgacagaagc ctcaagaaaa
     5941 aaaaaattct tcttcgacta tgctggaggc agagatgatc gagccggtag ttaactatat
     6001 atagctaaat tggttccatc acctcg
//
