LOCUS       sequence-198912-        5979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Estrogen inducing G-CSF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5979)
  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 5979)
  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: JC021
FEATURES             Location/Qualifiers
     source          1..5979
                     /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"
     rep_origin      complement(1404..1992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1484..1503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2112..2161
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     promoter        2282..2688
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2718..2738
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3957..4106
                     /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             4113..4139
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      4181..4427
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             join(4988..5044,5045..5242)
                     /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             5789..5818
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5834..5851
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      join(5875..5979,1..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 caacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1381 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1441 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1501 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1561 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     1621 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1741 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1801 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     1861 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1921 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1981 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2041 aactcacgtt aagggatttt ggtcatgaga tcagatctga gctcgtttgg ccgtggccgt
     2101 gctcgtcctc gtcggccggc ttgtcgacga cggcggtctc cgtcgtcagg atcatccggg
     2161 ccacaagctt gctgacagaa gcctcaagaa aaaaaaaatt cttcttcgac tatgctggag
     2221 gcagagatga tcgagccggt agttaactat atatagctaa attggttcca tcacctcgag
     2281 tccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
     2341 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
     2401 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
     2461 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
     2521 tcttgaaaat tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
     2581 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
     2641 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagga cggcaactag
     2701 taaaaatgcg cggagctcct aagaaaaagc gcaaagtccg gccggcatct agacccgggg
     2761 agcgcccctt ccagtgtcgc atttgcatgc ggaacttttc gcgccaggac aggcttgaca
     2821 ggcatacccg tactcatacc ggtgaaaaac cgtttcagtg tcggatctgt atgcgaaatt
     2881 tctcccagaa ggagcacttg gcggggcatc tacgtacgca caccggcgag aagccattcc
     2941 aatgccgaat atgcatgcgc aacttcagtc gccgcgacaa cctgaaccgg cacctaaaaa
     3001 cccacctgag gaacatatgc ggcggaggca cacctgcagc tgcgtcgact ctagaggatc
     3061 catctgctgg agacatgaga gctgccaacc tttggccaag cccgctcatg atcaaacgct
     3121 ctaagaagaa cagcctggcc ttgtccctga cggccgacca gatggtcagt gccttgttgg
     3181 atgctgagcc ccccatactc tattccgagt atgatcctac cagacccttc agtgaagctt
     3241 cgatgatggg cttactgacc aacctggcag acagggagct ggttcacatg atcaactggg
     3301 cgaagagggt gccaggcttt gtggatttga ccctccatga tcaggtccac cttctagaat
     3361 gtgcctggct agagatcctg atgattggtc tcgtctggcg ctccatggag cacccagtga
     3421 agctactgtt tgctcctaac ttgctcttgg acaggaacca gggaaaatgt gtagagggca
     3481 tggtggagat cttcgacatg ctgctggcta catcatctcg gttccgcatg atgaatctgc
     3541 agggagagga gtttgtgtgc ctcaaatcta ttattttgct taattctgga gtgtacacat
     3601 ttctgtccag caccctgaag tctctggaag agaaggacca tatccaccga gtcctggaca
     3661 agatcacaga cactttgatc cacctgatgg ccaaggcagg cctgaccctg cagcagcagc
     3721 accagcggct ggcccagctc ctcctcatcc tctcccacat caggcacatg agtaacaaag
     3781 gcatggagca tctgtacagc atgaagtgca agaacgtggt gcccctctat gacctgctgc
     3841 tggagatgct ggacgcccac cgcctacatg cgcccactag ccgtggaggg gcatccgtgg
     3901 aggagacgga ccaaagccac ttggccactg cgggctctac ttcatcgcct agggccgacg
     3961 cgctggacga tttcgatctc gacatgctgg gttctgatgc cctcgatgac tttgacctgg
     4021 atatgttggg aagcgacgca ttggatgact ttgatctgga catgctcggc tccgatgctc
     4081 tggacgattt cgatctcgat atgttaatta actacccgta cgacgttccg gactacgctt
     4141 cttgaggtac catcggtaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc
     4201 catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt
     4261 ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc
     4321 tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt
     4381 tcttggtatt tcccactcct cttcagagta cagaagatta agtgagagct agcatatcat
     4441 atagaagtca tcgcggcaga tcaattcatc aatcggagtg aggatattgc gagtttacca
     4501 ccatcaatcg gagtgaggat ctccaattgg tgacggtcca gtcatcaatc ggagtgagga
     4561 ttcagctgct tctcgaggcc gcacatcaat cggagtgagg atgtacaggg tgggctgttc
     4621 caccatcaat cggagtgagg atttgcgtca atggggcgga gttcatcaat cggagtgagg
     4681 atatcgaagt catcgagagc actcatcaat cggagtgagg atatactcca cccattgacg
     4741 tcacatcaat cggagtgagg attggaacca gaagggtctc ttcatcaatc ggagtgagga
     4801 taccctatgg gcgcgccgtc gctggcaata atagcgggcg gacgcatgtc atgagattat
     4861 tggaaaccac cagaatcgaa tataaaaggc gaacaccttt cccaattttg gtttcccctg
     4921 acccaaagac tttaaattta atttatttgt ccctatttca atcaattgaa caactatcaa
     4981 aacacagatg agatttcctt caatttttac tgctgtttta ttcgcagcat cctccgcatt
     5041 agctgctcca gtcaacacta caacagaaga tgaaacggca caaattccgg ctgaagctgt
     5101 catcggttac tcagatttag aaggggattt cgatgttgct gttttgccat tttccaacag
     5161 cacaaataac gggttattgt ttataaatac tactattgcc agcattgctg ctaaagaaga
     5221 aggggtatct ctcgagaaga gaatgactcc tttgggtcca gcttcttcct tgcctcaatc
     5281 cttcttgttg aagtgtttgg agcaggttag aaagatccag ggtgatggtg ctgctttgca
     5341 agagaagttg tgtgctactt acaagttgtg tcacccagaa gagttggttt tgttgggtca
     5401 ctccttgggt attccttggg ctccattgtc ctcttgtcca tcccaagctt tgcaattggc
     5461 tggttgtttg tcccaattgc actccggttt gttcttgtac cagggtttgt tgcaagcttt
     5521 ggagggtatt tctccagagt tgggtccaac tttggacaca ttgcagttgg acgttgctga
     5581 cttcgctact actatctggc aacagatgga agaattgggt atggctccag ctttgcagcc
     5641 aactcaaggt gctatgccag cttttgcttc tgctttccag agaagagctg gtggtgtttt
     5701 ggttgcttct cacttgcagt ctttcttgga ggtttcctac agagttttga gacacttggc
     5761 tcaaccataa gcggccgcca gctttctaga acaaaaactc atctcagaag aggatctgaa
     5821 tagcgccgtc gaccatcatc atcatcatca ttgaccggtc ttgctagatt ctaatcaaga
     5881 ggatgtcaga atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt
     5941 gtaacctata tagtatagga ttttttttgt cattttgtt
//
