LOCUS       sequence-147733-        3640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supplemental Figure 4. Promoter short ppTEF1. Expresses GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3640)
  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 3640)
  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: PP164
FEATURES             Location/Qualifiers
     source          1..3640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      279..526
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(295..313)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(601..1189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(681..700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1309..1358
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     CDS             join(1906..1908,1909..1911,1912..2622)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1906..1908,1909..1911,1912..2622)
                     /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(1951..1972)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2212..2231)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2559..2580
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2647..2676
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2692..2709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2733..2979
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2994..3405
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3413..3460
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(3479..3640,1..213)
                     /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 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
       61 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
      121 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
      181 tgcgtgcact tcgtggccga ggagcaggac tgacacgtcc gacggcggcc cacgggtccc
      241 aggcctcgga gatccgtccc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
      301 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
      361 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
      421 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
      481 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc tggagaccaa
      541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      781 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1021 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     1081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1201 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1261 catgagatca gatctgagct cgtttggccg tggccgtgct cgtcctcgtc ggccggcttg
     1321 tcgacgacgg cggtctccgt cgtcaggatc atccgggcca caagcttgct gacagaagcc
     1381 tcaagaaaaa aaaaattctt cttcgactat gctggaggca gagatgatcg agccggtagt
     1441 taactatata tagctaaatt ggttccatca cctcgaataa ctgtcgcctc ttttatctgc
     1501 cgcactgcat gaggtgtccc cttagtggga aagagtactg agccaaccct ggaggacagc
     1561 aagggaaaaa tacctacaac ttgcttcata atggtcgtaa aaacaatcct tgtcggatat
     1621 aagtgttgta gactgtccct tatcctctgc gatgttcttc ctctcaaagt ttgcgatttc
     1681 tctctatcag aattgccatc aagagactca ggactaattt cgcagtccca cacgcactcg
     1741 tacatgattg gctgaaattt ccctaaagaa tttctttttc acgaaaattt ttttttacac
     1801 aagattttca gcagatataa aatggagagc aggacctccg ctgtgactct tctttttttt
     1861 cttttattct cactacatac attttagtta ttcgccaacg aattcatggt gagcaagggc
     1921 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1981 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2041 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2101 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2161 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2221 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2281 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2341 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2401 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2461 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     2521 agcgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2581 accgccgccg ggatcactct cggcatggac gagctgtaca aggccgcggc ggccgccagc
     2641 tttctagaac aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat
     2701 catcatcatt gaccggtctt gctagattct aatcaagagg atgtcagaat gccatttgcc
     2761 tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata gtataggatt
     2821 ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta tctcgcagct
     2881 gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt tttcttggta
     2941 tttcccactc ctcttcagag tacagaagat taagtgagac cttcgtttgt gcggatcccc
     3001 cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac
     3061 tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat tttccctctt
     3121 tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc
     3181 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc
     3241 ttgaaatttt tttttttagt ttttttctct ttcagtgacc tccattgata tttaagttaa
     3301 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
     3361 ttacttcttg ttcattagaa agaaagcata gcaatctaat ctaaggggcg gtgttgacaa
     3421 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     3481 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
     3541 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
     3601 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
//
