LOCUS       sequence-147732-        3623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supplemental Figure 4. Promoter scTEF1. Expresses GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3623)
  AUTHORS   Perez-Pinera P, Han N, Cleto S, Cao J, Purcell O, Shah KA, Lee K, 
            Ram R, Lu TK
  TITLE     Synthetic biology and microbioreactor platforms for programmable 
            production of biologics at the point-of-care.
  JOURNAL   Nat Commun. 2016 Jul 29;7:12211. doi: 10.1038/ncomms12211.
  PUBMED    27470089
REFERENCE   2  (bases 1 to 3623)
  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: "2016-07-29"; volume: "7:12211. doi"; pages: " 
            10.1038/ncomms12211"
COMMENT     Sequence Label: PP87
FEATURES             Location/Qualifiers
     source          1..3623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      257..504
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(273..291)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(579..1167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(659..678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1287..1336
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     promoter        1456..1860
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             join(1867..1869,1870..1872,1873..2583)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1867..1869,1870..1872,1873..2583)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1912..1933)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2173..2192)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2520..2541
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2608..2637
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2653..2670
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2694..2940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2955..3366
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3374..3421
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3440..3623,1..191)
                     /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"
ORIGIN
        1 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
       61 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
      121 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
      181 agcaggactg acacgtccga cggcggccca cgggtcccag gcctcggaga tccgtccccc
      241 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
      301 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
      361 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
      421 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
      481 ggacgctcga aggctttaat ttgcaagctg gagaccaaca tgtgagcaaa aggccagcaa
      541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
      781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     1021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagatcaga tctgagctcg
     1261 tttggccgtg gccgtgctcg tcctcgtcgg ccggcttgtc gacgacggcg gtctccgtcg
     1321 tcaggatcat ccgggccaca agcttgctga cagaagcctc aagaaaaaaa aaattcttct
     1381 tcgactatgc tggaggcaga gatgatcgag ccggtagtta actatatata gctaaattgg
     1441 ttccatcacc tcgagccaca caccatagct tcaaaatgtt tctactcctt ttttactctt
     1501 ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata
     1561 ctaaattttc cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga
     1621 aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt
     1681 atcacgtttc tttttcttga aatttttttt tttagttttt ttctctttca gtgacctcca
     1741 ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt
     1801 tctattacaa ctttttttac ttcttgttca ttagaaagaa acatagcaat ctaatctaag
     1861 gaattcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1921 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1981 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2041 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2101 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2161 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2221 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2281 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2341 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2401 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2461 aaccactacc tgagcaccca gagcgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2521 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2581 aaggccgcgg cggccgccag ctttctagaa caaaaactca tctcagaaga ggatctgaat
     2641 agcgccgtcg accatcatca tcatcatcat tgaccggtct tgctagattc taatcaagag
     2701 gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact tttttatttg
     2761 taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg agcttgctcc
     2821 tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga aaatcattcg
     2881 agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga ttaagtgaga
     2941 ccttcgtttg tgcggatccc ccacacacca tagcttcaaa atgtttctac tcctttttta
     3001 ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca
     3061 gcatactaaa ttttccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt
     3121 ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa
     3181 tttttatcac gtttcttttt cttgaaattt ttttttttag tttttttctc tttcagtgac
     3241 ctccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt
     3301 cttgttctat tacaactttt tttacttctt gttcattaga aagaaagcat agcaatctaa
     3361 tctaaggggc ggtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3421 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
     3481 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
     3541 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
     3601 ggaccaggtg gtgccggaca aca
//
