LOCUS       sequence-203154-        5974 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Estrogen inducing G-CSF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5974)
  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 5974)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..5974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..423
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             447..467
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1686..1835
                     /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             1842..1868
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      1910..2156
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             join(2717..2773,2774..2971)
                     /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             3518..3547
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3563..3580
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3604..3850
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3865..4278
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        4285..4332
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4351..4725
                     /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      4791..5038
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4807..4825)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(5113..5701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5193..5212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5821..5870
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
ORIGIN
        1 ttccatcacc tcgagtccac acaccatagc ttcaaaatgt ttctactcct tttttactct
       61 tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat
      121 actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg
      181 aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt
      241 tatcacgttt ctttttcttg aaaatttttt ttttgatttt tttctctttc gatgacctcc
      301 cattgatatt taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt
      361 gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct
      421 aaggactagt aaaaatgcgc ggagctccta agaaaaagcg caaagtccgg ccggcatcta
      481 gacccgggga gcgccccttc cagtgtcgca tttgcatgcg gaacttttcg cgccaggaca
      541 ggcttgacag gcatacccgt actcataccg gtgaaaaacc gtttcagtgt cggatctgta
      601 tgcgaaattt ctcccagaag gagcacttgg cggggcatct acgtacgcac accggcgaga
      661 agccattcca atgccgaata tgcatgcgca acttcagtcg ccgcgacaac ctgaaccggc
      721 acctaaaaac ccacctgagg aacatatgcg gcggaggcac acctgcagct gcgtcgactc
      781 tagaggatcc atctgctgga gacatgagag ctgccaacct ttggccaagc ccgctcatga
      841 tcaaacgctc taagaagaac agcctggcct tgtccctgac ggccgaccag atggtcagtg
      901 ccttgttgga tgctgagccc cccatactct attccgagta tgatcctacc agacccttca
      961 gtgaagcttc gatgatgggc ttactgacca acctggcaga cagggagctg gttcacatga
     1021 tcaactgggc gaagagggtg ccaggctttg tggatttgac cctccatgat caggtccacc
     1081 ttctagaatg tgcctggcta gagatcctga tgattggtct cgtctggcgc tccatggagc
     1141 acccagtgaa gctactgttt gctcctaact tgctcttgga caggaaccag ggaaaatgtg
     1201 tagagggcat ggtggagatc ttcgacatgc tgctggctac atcatctcgg ttccgcatga
     1261 tgaatctgca gggagaggag tttgtgtgcc tcaaatctat tattttgctt aattctggag
     1321 tgtacacatt tctgtccagc accctgaagt ctctggaaga gaaggaccat atccaccgag
     1381 tcctggacaa gatcacagac actttgatcc acctgatggc caaggcaggc ctgaccctgc
     1441 agcagcagca ccagcggctg gcccagctcc tcctcatcct ctcccacatc aggcacatga
     1501 gtaacaaagg catggagcat ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg
     1561 acctgctgct ggagatgctg gacgcccacc gcctacatgc gcccactagc cgtggagggg
     1621 catccgtgga ggagacggac caaagccact tggccactgc gggctctact tcatcgccta
     1681 gggccgacgc gctggacgat ttcgatctcg acatgctggg ttctgatgcc ctcgatgact
     1741 ttgacctgga tatgttggga agcgacgcat tggatgactt tgatctggac atgctcggct
     1801 ccgatgctct ggacgatttc gatctcgata tgttaattaa ctacccgtac gacgttccgg
     1861 actacgcttc ttgaggtacc atcggtagac cggtcttgct agattctaat caagaggatg
     1921 tcagaatgcc atttgcctga gagatgcagg cttcattttt gatacttttt tatttgtaac
     1981 ctatatagta taggattttt tttgtcattt tgtttcttct cgtacgagct tgctcctgat
     2041 cagcctatct cgcagctgat gaatatcttg tggtaggggt ttgggaaaat cattcgagtt
     2101 tgatgttttt cttggtattt cccactcctc ttcagagtac agaagattaa gtgagagcta
     2161 gcatatcata tagaagtcat cgcggcagat caattcatca atcggagtga ggatattgcg
     2221 agtttaccac catcaatcgg agtgaggatc tccaattggt gacggtccag tcatcaatcg
     2281 gagtgaggat tcagctgctt ctcgaggccg cacatcaatc ggagtgagga tgtacagggt
     2341 gggctgttcc accatcaatc ggagtgagga tttgcgtcaa tggggcggag ttcatcaatc
     2401 ggagtgagga tatcgaagtc atcgagagca ctcatcaatc ggagtgagga tatactccac
     2461 ccattgacgt cacatcaatc ggagtgagga ttggaaccag aagggtctct tcatcaatcg
     2521 gagtgaggat accctatggg cgcgccgtcg ctggcaataa tagcgggcgg acgcatgtca
     2581 tgagattatt ggaaaccacc agaatcgaat ataaaaggcg aacacctttc ccaattttgg
     2641 tttcccctga cccaaagact ttaaatttaa tttatttgtc cctatttcaa tcaattgaac
     2701 aactatcaaa acacagatga gatttccttc aatttttact gctgttttat tcgcagcatc
     2761 ctccgcatta gctgctccag tcaacactac aacagaagat gaaacggcac aaattccggc
     2821 tgaagctgtc atcggttact cagatttaga aggggatttc gatgttgctg ttttgccatt
     2881 ttccaacagc acaaataacg ggttattgtt tataaatact actattgcca gcattgctgc
     2941 taaagaagaa ggggtatctc tcgagaagag aatgactcct ttgggtccag cttcttcctt
     3001 gcctcaatcc ttcttgttga agtgtttgga gcaggttaga aagatccagg gtgatggtgc
     3061 tgctttgcaa gagaagttgt gtgctactta caagttgtgt cacccagaag agttggtttt
     3121 gttgggtcac tccttgggta ttccttgggc tccattgtcc tcttgtccat cccaagcttt
     3181 gcaattggct ggttgtttgt cccaattgca ctccggtttg ttcttgtacc agggtttgtt
     3241 gcaagctttg gagggtattt ctccagagtt gggtccaact ttggacacat tgcagttgga
     3301 cgttgctgac ttcgctacta ctatctggca acagatggaa gaattgggta tggctccagc
     3361 tttgcagcca actcaaggtg ctatgccagc ttttgcttct gctttccaga gaagagctgg
     3421 tggtgttttg gttgcttctc acttgcagtc tttcttggag gtttcctaca gagttttgag
     3481 acacttggct caaccataag cggccgccag ctttctagaa caaaaactca tctcagaaga
     3541 ggatctgaat agcgccgtcg accatcatca tcatcatcat tgaccggtct tgctagattc
     3601 taatcaagag gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact
     3661 tttttatttg taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg
     3721 agcttgctcc tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga
     3781 aaatcattcg agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga
     3841 ttaagtgaga ccttcgtttg tgcggatccc ccacacacca tagcttcaaa atgtttctac
     3901 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     3961 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     4021 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     4081 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct
     4141 ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt ctcaagtttc
     4201 agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc
     4261 atagcaatct aatctaaggg cggtgttgac aattaatcat cggcatagta tatcggcata
     4321 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     4381 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     4441 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     4501 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     4561 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     4621 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     4681 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt ccgacgcggc
     4741 ccgacgggtc cgaggcctcg gagatccgtc ccccttttcc tttgtcgata tcatgtaatt
     4801 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     4861 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     4921 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     4981 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa
     5041 gctggagacc aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5161 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5221 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5281 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5341 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5401 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5461 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5521 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5581 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5641 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5761 agggattttg gtcatgagat cagatctgag ctcgtttggc cgtggccgtg ctcgtcctcg
     5821 tcggccggct tgtcgacgac ggcggtctcc gtcgtcagga tcatccgggc cacaagcttg
     5881 ctgacagaag cctcaagaaa aaaaaaattc ttcttcgact atgctggagg cagagatgat
     5941 cgagccggta gttaactata tatagctaaa ttgg
//
