LOCUS       sequence-369680-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Characterisation of yeast 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 5057)
  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 5057)
  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: pILGFP1D5
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(287..303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(287..303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(300..322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    311..327
                     /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(335..341,342..359,360..365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    380..401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(518..535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(689..1277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(769..788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1448..2239,2240..2308))
                     /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     2071..2090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2309..2413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2481..2499
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2537..2559)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2659..2678
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2872..2894
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2886..2903
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2887..2903
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4017..4050)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             4089..4799
                     /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      join(4809..5057,1..22)
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
ORIGIN
        1 aataactcga aaattctgcg ttcgttaaag ctttcgagaa ggatattatt tactagtagg
       61 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
      121 ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc
      181 atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgggaa
      241 tctcggtcgt aatgatttaa atgcatgcaa gcttggcgta atcatggtca tagctgtttc
      301 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      361 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      421 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     2461 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     2521 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2581 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2641 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     2701 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     2761 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     2821 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     2881 gacgttgtaa aacgacggcc agtgaattca tttaaatttt tttgattcgg taatctccga
     2941 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
     3001 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacctg
     3061 caggaaacga agataaaaca tgtccacaaa atcatatacc agtagagctg agactcatgc
     3121 aagtccggtt gcatcgaaac ttttacgttt aatggatgaa aagaagacca atttgtgtgc
     3181 ttctcttgac gttcgttcga ctgatgagct attgaaactt gttgaaacgt tgggtccata
     3241 catttgcctt ttgaaaacac acgttgatat cttggatgat ttcagttatg agggtactgt
     3301 cgttccattg aaagcattgg cagagaaata caagttcttg atatttgagg acagaaaatt
     3361 cgccgatatc ggtaacacag tcaaattaca atatacatcg ggcgtttacc gtatcgcaga
     3421 atggtctgat atcaccaacg cccacggggt tactggtgct ggtattgttg ctggcttgaa
     3481 acaaggtgcg caagaggtca ccaaagaacc aaggggatta ttgatgcttg ctgaattgtc
     3541 ttccaagggt tctctagcac acggtgaata tactaagggt accgttgata ttgcaaagag
     3601 tgataaagat ttcgttattg ggttcattgc tcagaacgat atgggaggaa gagaagaagg
     3661 gtttgattgg ctaatcatga ccccaggtgt aggtttagac gacaaaggcg atgcattggg
     3721 tcagcagtac agaaccgtcg acgaagttgt aagtggtgga tcagatatca tcattgttgg
     3781 cagaggactt ttcgccaagg gtagagatcc taaggttgaa ggtgaaagat acagaaatgc
     3841 tggatgggaa gcgtaccaaa agagaatcag cgctccccat taattataca ggaaacttaa
     3901 tagaacaaat cacatattta atctaatagc cacctgcatt ggcacggtgc aacactactt
     3961 caacttcatc ttacaaaaag atcacgtgat ctgttgtatt gggatctcta gacctaataa
     4021 cttcgtatag catacattat acgaagttat attaagggtt gctcgagaaa gagctcaaaa
     4081 tgggatcctc taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag
     4141 atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt
     4201 acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa
     4261 ccttagtcac tactttcggt tatggtgttc aatgttttgc gagataccca gatcatatga
     4321 aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt
     4381 ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct
     4441 tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc
     4501 acaaattgga atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga
     4561 atggtatcaa agttaacttc aaaattagac acaacattga agatggttct gttcaattag
     4621 ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc
     4681 attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg
     4741 tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaaa
     4801 gatcttaaat tgaattgaat tgaaatcgat agatcaattt ttttcttttc tctttcccca
     4861 tcctttacgc taaaataata gtttatttta ttttttgaat attttttatt tatatacgta
     4921 tatatagact attatttatc ttttaatgat tattaagatt tttattaaaa aaaaattcgc
     4981 tcctctttta atgcctttat gcagtttttt tttcccattc gatatttcta tgttcgggtt
     5041 cagcgtattt taagttt
//
