LOCUS       sequence-369628-        5453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the CYC1 core promoter inserted with 4 Z268 elements using a
            green fluorescence protein as the reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5453)
  AUTHORS   Peng B, Bandari NC, Lu Z, Howard CB, Scott C, Trau M, Dumsday G, 
            Vickers CE
  TITLE     Engineering eukaryote-like regulatory circuits to expand artificial 
            control mechanisms for metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Commun Biol. 2022 Feb 16;5(1):135. doi: 10.1038/s42003-022-03070-z.
  PUBMED    35173283
REFERENCE   2  (bases 1 to 5453)
  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: "Commun 
            Biol."; date: "2022-02-16"; volume: "5(1):135. doi"; pages: " 
            10.1038/s42003-022-03070-z"
COMMENT     Sequence Label: pILGFP4D41
FEATURES             Location/Qualifiers
     source          1..5453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1998..2016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2054..2076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2176..2195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2389..2411
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2403..2420
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2404..2420
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3534..3567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3999..4712
                     /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      4722..4992
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(5257..5273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5257..5273)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5270..5292)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5281..5297
                     /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(5305..5311,5312..5329,5330..5335))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5350..5371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2221 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     2281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2341 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2401 gttgtaaaac gacggccagt gaattcattt aaattttttt gattcggtaa tctccgaaca
     2461 gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt
     2521 tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag
     2581 gaaacgaaga taaaacatgt ccacaaaatc atataccagt agagctgaga ctcatgcaag
     2641 tccggttgca tcgaaacttt tacgtttaat ggatgaaaag aagaccaatt tgtgtgcttc
     2701 tcttgacgtt cgttcgactg atgagctatt gaaacttgtt gaaacgttgg gtccatacat
     2761 ttgccttttg aaaacacacg ttgatatctt ggatgatttc agttatgagg gtactgtcgt
     2821 tccattgaaa gcattggcag agaaatacaa gttcttgata tttgaggaca gaaaattcgc
     2881 cgatatcggt aacacagtca aattacaata tacatcgggc gtttaccgta tcgcagaatg
     2941 gtctgatatc accaacgccc acggggttac tggtgctggt attgttgctg gcttgaaaca
     3001 aggtgcgcaa gaggtcacca aagaaccaag gggattattg atgcttgctg aattgtcttc
     3061 caagggttct ctagcacacg gtgaatatac taagggtacc gttgatattg caaagagtga
     3121 taaagatttc gttattgggt tcattgctca gaacgatatg ggaggaagag aagaagggtt
     3181 tgattggcta atcatgaccc caggtgtagg tttagacgac aaaggcgatg cattgggtca
     3241 gcagtacaga accgtcgacg aagttgtaag tggtggatca gatatcatca ttgttggcag
     3301 aggacttttc gccaagggta gagatcctaa ggttgaaggt gaaagataca gaaatgctgg
     3361 atgggaagcg taccaaaaga gaatcagcgc tccccattaa ttatacagga aacttaatag
     3421 aacaaatcac atatttaatc taatagccac ctgcattggc acggtgcaac actacttcaa
     3481 cttcatctta caaaaagatc acgtgatctg ttgtattggg atctctagac ctaataactt
     3541 cgtatagcat acattatacg aagttatatt aagggttgct cgagaaagag ctcacggcgg
     3601 ccaagcacgc ggggataatg aactagattt ccgtgtgaga cgacatcgtc gatactgatg
     3661 taatcaagtt cctacctgaa tctaagattg ggcccgcgtg ggcgaattgg tgcgtgggcg
     3721 taagcggggg cgggcgtggg cgtagcgtgg gcggccggca atcctcgagc agatccgcca
     3781 ggcgtgtata tatagcgtgg atggccaggc aactttagtg ctgacacata caggcatata
     3841 tatatgtgtg cgacgacaca tgatcatatg gcatgcatgt gctctgtatg tatataaaac
     3901 tcttgttttc ttcttttctc taaatattct ttccttatac attaggacct ttgcagcata
     3961 aattactata cttctataga cacacaaacg gatccaaaat gtctaaaggt gaagaattat
     4021 tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt
     4081 ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt
     4141 gtactactgg taaattgcca gttccatggc caaccttagt cactactttc ggttatggtg
     4201 ttcaatgttt tgcgagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca
     4261 tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga
     4321 ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta
     4381 ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc
     4441 acaatgttta catcatggct gacaaacaaa agaatggtat caaagttaac ttcaaaatta
     4501 gacacaacat tgaagatggt tctgttcaat tagctgacca ttatcaacaa aatactccaa
     4561 ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa tctgccttat
     4621 ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg
     4681 gtattaccca tggtatggat gaattgtaca aaagatctta aattgaattg aattgaaatc
     4741 gatagatcaa tttttttctt ttctctttcc ccatccttta cgctaaaata atagtttatt
     4801 ttattttttg aatatttttt atttatatac gtatatatag actattattt atcttttaat
     4861 gattattaag atttttatta aaaaaaaatt cgctcctctt ttaatgcctt tatgcagttt
     4921 ttttttccca ttcgatattt ctatgttcgg gttcagcgta ttttaagttt aataactcga
     4981 aaattctgcg ttcgttaaag ctttcgagaa ggatattatt tactagtagg actatttgca
     5041 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     5101 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     5161 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgggaa tctcggtcgt
     5221 aatgatttaa atgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5281 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5341 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5401 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg gga
//
