LOCUS       sequence-327687-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Characterisation of TEF1 promoter using yEGFP as reporter gene and 
            PGK1 terminator. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  AUTHORS   Lu Z, Peng B, Ebert BE, Dumsday G, Vickers CE
  TITLE     Auxin-mediated protein depletion for metabolic engineering in 
            terpene-producing yeast.
  JOURNAL   Nat Commun. 2021 Feb 16;12(1):1051. doi: 10.1038/s41467-021-21313-1.
  PUBMED    33594068
REFERENCE   2  (bases 1 to 5668)
  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: "2021-02-16"; volume: "12(1):1051. doi"; pages: " 
            10.1038/s41467-021-21313-1"
COMMENT     Sequence Label: pILGFP1H5
FEATURES             Location/Qualifiers
     source          1..5668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             422..1135
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1145..1415
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(1680..1696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1680..1696)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1693..1715)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1704..1720
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1728..1734,1735..1752,1753..1758))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1773..1794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1911..1928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2082..2670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2162..2181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2841..3632,3633..3701))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3464..3483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3702..3806)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3874..3892
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3930..3952)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4052..4071
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4265..4287
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4279..4296
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4280..4296
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5410..5443)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(5663..5668,1..408)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga
       61 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct
      121 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac
      181 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt
      241 cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
      301 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
      361 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa
      421 aatgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg aattagatgg
      481 tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg
      541 taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat ggccaacctt
      601 agtcactact ttcggttatg gtgttcaatg ttttgcgaga tacccagatc atatgaaaca
      661 acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa ctattttttt
      721 caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg ataccttagt
      781 taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt taggtcacaa
      841 attggaatac aactataact ctcacaatgt ttacatcatg gctgacaaac aaaagaatgg
      901 tatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggttctgttc aattagctga
      961 ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag acaaccatta
     1021 cttatccact caatctgcct tatccaaaga tccaaacgaa aagagagacc acatggtctt
     1081 gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattgt acaaaagatc
     1141 ttaaattgaa ttgaattgaa atcgatagat caattttttt cttttctctt tccccatcct
     1201 ttacgctaaa ataatagttt attttatttt ttgaatattt tttatttata tacgtatata
     1261 tagactatta tttatctttt aatgattatt aagattttta ttaaaaaaaa attcgctcct
     1321 cttttaatgc ctttatgcag tttttttttc ccattcgata tttctatgtt cgggttcagc
     1381 gtattttaag tttaataact cgaaaattct gcgttcgtta aagctttcga gaaggatatt
     1441 atttactagt aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     1501 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     1561 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     1621 tattacccgg gaatctcggt cgtaatgatt taaatgcatg caagcttggc gtaatcatgg
     1681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3841 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3901 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3961 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4021 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     4081 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     4141 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     4201 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     4261 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcatttaaat ttttttgatt
     4321 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     4381 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     4441 gaacaaaaac ctgcaggaaa cgaagataaa acatgtccac aaaatcatat accagtagag
     4501 ctgagactca tgcaagtccg gttgcatcga aacttttacg tttaatggat gaaaagaaga
     4561 ccaatttgtg tgcttctctt gacgttcgtt cgactgatga gctattgaaa cttgttgaaa
     4621 cgttgggtcc atacatttgc cttttgaaaa cacacgttga tatcttggat gatttcagtt
     4681 atgagggtac tgtcgttcca ttgaaagcat tggcagagaa atacaagttc ttgatatttg
     4741 aggacagaaa attcgccgat atcggtaaca cagtcaaatt acaatataca tcgggcgttt
     4801 accgtatcgc agaatggtct gatatcacca acgcccacgg ggttactggt gctggtattg
     4861 ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga accaagggga ttattgatgc
     4921 ttgctgaatt gtcttccaag ggttctctag cacacggtga atatactaag ggtaccgttg
     4981 atattgcaaa gagtgataaa gatttcgtta ttgggttcat tgctcagaac gatatgggag
     5041 gaagagaaga agggtttgat tggctaatca tgaccccagg tgtaggttta gacgacaaag
     5101 gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt tgtaagtggt ggatcagata
     5161 tcatcattgt tggcagagga cttttcgcca agggtagaga tcctaaggtt gaaggtgaaa
     5221 gatacagaaa tgctggatgg gaagcgtacc aaaagagaat cagcgctccc cattaattat
     5281 acaggaaact taatagaaca aatcacatat ttaatctaat agccacctgc attggcacgg
     5341 tgcaacacta cttcaacttc atcttacaaa aagatcacgt gatctgttgt attgggatct
     5401 ctagacctaa taacttcgta tagcatacat tatacgaagt tatattaagg gttgctcgag
     5461 aaagagctcc agaaagcgac cacccaactt tggctgataa tagcgtataa acaatgcata
     5521 ctttgtacgt tcaaaataca atgcagtaga tatatttatg catattacat ataatacata
     5581 tcacatagga agcaacaggc gcgttggact tttaattttc gaggaccgcg aatccttaca
     5641 tcacacccaa tcccccacaa gtgatccc
//
