LOCUS       sequence-327686-        5674 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 5674)
  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 5674)
  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: pILGFP1F5
FEATURES             Location/Qualifiers
     source          1..5674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        217..630
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             653..1363
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      1373..1643
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(1908..1924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1908..1924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1921..1943)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1932..1948
                     /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(1956..1962,1963..1980,1981..1986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2001..2022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2139..2156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2310..2898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2390..2409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3069..3860,3861..3929))
                     /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     3692..3711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3930..4034)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4102..4120
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4158..4180)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4280..4299
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4493..4515
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4507..4524
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4508..4524
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5638..5671)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 aagggttgct cgagaaagag ctccagaaag cgaccaccca actttggctg ataatagcgt
       61 ataaacaatg catactttgt acgttcaaaa tacaatgcag tagatatatt tatgcatatt
      121 acatataata catatcacat aggaagcaac aggcgcgttg gacttttaat tttcgaggac
      181 cgcgaatcct tacatcacac ccaatccccc acaagtgatc ccccacacac catagcttca
      241 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
      301 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
      361 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
      421 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
      481 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat
      541 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
      601 tagaaagaaa gcatagcaat ctaatctaag ttttaattac aaaatgggat cctctaaagg
      661 tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg
      721 tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt
      781 aaaatttatt tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt
      841 cggttatggt gttcaatgtt ttgcgagata cccagatcat atgaaacaac atgacttttt
      901 caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg
      961 taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga
     1021 attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa
     1081 ctataactct cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa
     1141 cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca
     1201 aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact tatccactca
     1261 atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt
     1321 tactgctgct ggtattaccc atggtatgga tgaattgtac aaaagatctt aaattgaatt
     1381 gaattgaaat cgatagatca atttttttct tttctctttc cccatccttt acgctaaaat
     1441 aatagtttat tttatttttt gaatattttt tatttatata cgtatatata gactattatt
     1501 tatcttttaa tgattattaa gatttttatt aaaaaaaaat tcgctcctct tttaatgcct
     1561 ttatgcagtt tttttttccc attcgatatt tctatgttcg ggttcagcgt attttaagtt
     1621 taataactcg aaaattctgc gttcgttaaa gctttcgaga aggatattat ttactagtag
     1681 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     1741 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     1801 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccggga
     1861 atctcggtcg taatgattta aatgcatgca agcttggcgt aatcatggtc atagctgttt
     1921 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1981 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2101 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     2161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     2221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4081 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     4141 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4201 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4261 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     4321 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     4381 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     4441 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     4501 cgacgttgta aaacgacggc cagtgaattc atttaaattt ttttgattcg gtaatctccg
     4561 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     4621 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacct
     4681 gcaggaaacg aagataaaac atgtccacaa aatcatatac cagtagagct gagactcatg
     4741 caagtccggt tgcatcgaaa cttttacgtt taatggatga aaagaagacc aatttgtgtg
     4801 cttctcttga cgttcgttcg actgatgagc tattgaaact tgttgaaacg ttgggtccat
     4861 acatttgcct tttgaaaaca cacgttgata tcttggatga tttcagttat gagggtactg
     4921 tcgttccatt gaaagcattg gcagagaaat acaagttctt gatatttgag gacagaaaat
     4981 tcgccgatat cggtaacaca gtcaaattac aatatacatc gggcgtttac cgtatcgcag
     5041 aatggtctga tatcaccaac gcccacgggg ttactggtgc tggtattgtt gctggcttga
     5101 aacaaggtgc gcaagaggtc accaaagaac caaggggatt attgatgctt gctgaattgt
     5161 cttccaaggg ttctctagca cacggtgaat atactaaggg taccgttgat attgcaaaga
     5221 gtgataaaga tttcgttatt gggttcattg ctcagaacga tatgggagga agagaagaag
     5281 ggtttgattg gctaatcatg accccaggtg taggtttaga cgacaaaggc gatgcattgg
     5341 gtcagcagta cagaaccgtc gacgaagttg taagtggtgg atcagatatc atcattgttg
     5401 gcagaggact tttcgccaag ggtagagatc ctaaggttga aggtgaaaga tacagaaatg
     5461 ctggatggga agcgtaccaa aagagaatca gcgctcccca ttaattatac aggaaactta
     5521 atagaacaaa tcacatattt aatctaatag ccacctgcat tggcacggtg caacactact
     5581 tcaacttcat cttacaaaaa gatcacgtga tctgttgtat tgggatctct agacctaata
     5641 acttcgtata gcatacatta tacgaagtta tatt
//
