LOCUS       sequence-370793-        5449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the SkGAL2 promoter inserted with 2 PZ4 elements using a 
            green fluorescent protein as the reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5449)
  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 5449)
  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: pILGFP10E1A
FEATURES             Location/Qualifiers
     source          1..5449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             721..1431
                     /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"
     primer_bind     complement(1676..1692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1676..1692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1689..1711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1700..1716
                     /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(1724..1730,1731..1748,1749..1754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1769..1790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1907..1924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2078..2666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2158..2177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2837..3628,3629..3697))
                     /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     3460..3479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3698..3802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3870..3888
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3926..3948)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4048..4067
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4261..4283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4275..4292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4276..4292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5406..5439)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gctcgagtaa accaatttta tttgaacttg ccccgctaat cgtacagacg taccagatcc
       61 aggtcataac ggtgccatta gaaggccgta gaatagaccc actaaagcta tcacaggcaa
      121 ccttgcatcc aagtctaatt aaacggagca gaatctgttc cagtaatgta taaagaatta
      181 actaccggaa aagcgtcttc cggggcggct caaccggatt acattctttt ttcttttttg
      241 cggccagcat tcgagtcacg tgctcaaata ctgaggaagg ggttcttccg gtaggcaccg
      301 gcgctcactc ttccgtgcgg cacactcccc gcagttcagg ggccatgtgc ctgtgagagt
      361 atttccgcag gccatagttc cggggtggac cactccgagc taatgctaaa gtaagccctt
      421 cccaacttga aataggaaca aaagtcaatt atgctcctcc tgtaaagcta aactagagtg
      481 agacgttttt agagtgagac gttgttagca gtcgttgttc aagggaggga tgagttgagg
      541 aataaaatgc ggaaaaagtg tgggttagac agacatatat aaactctagc cattcggctg
      601 aaatatatct gagttcattt gtgataactt ttttgattac ttgttcatat atccacaaga
      661 acaacgacta ttaaacaata gaaaatagta taataaaaaa agttttttac aatgggatcc
      721 tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat
      781 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa
      841 ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc
      901 actactttcg gttatggtgt tcaatgtttt gcgagatacc cagatcatat gaaacaacat
      961 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
     1021 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
     1081 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
     1141 gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc
     1201 aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat
     1261 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
     1321 tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta
     1381 gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa aagatcttaa
     1441 actagtagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
     1501 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
     1561 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
     1621 acccgggaat ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa tcatggtcat
     1681 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1741 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1801 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1861 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2821 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2941 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3121 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3421 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3661 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3841 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     3901 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3961 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4021 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4081 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     4141 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4201 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4261 cccagtcacg acgttgtaaa acgacggcca gtgaattcat ttaaattttt ttgattcggt
     4321 aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg
     4381 catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac
     4441 aaaaacctgc aggaaacgaa gataaaacat gtccacaaaa tcatatacca gtagagctga
     4501 gactcatgca agtccggttg catcgaaact tttacgttta atggatgaaa agaagaccaa
     4561 tttgtgtgct tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt
     4621 gggtccatac atttgccttt tgaaaacaca cgttgatatc ttggatgatt tcagttatga
     4681 gggtactgtc gttccattga aagcattggc agagaaatac aagttcttga tatttgagga
     4741 cagaaaattc gccgatatcg gtaacacagt caaattacaa tatacatcgg gcgtttaccg
     4801 tatcgcagaa tggtctgata tcaccaacgc ccacggggtt actggtgctg gtattgttgc
     4861 tggcttgaaa caaggtgcgc aagaggtcac caaagaacca aggggattat tgatgcttgc
     4921 tgaattgtct tccaagggtt ctctagcaca cggtgaatat actaagggta ccgttgatat
     4981 tgcaaagagt gataaagatt tcgttattgg gttcattgct cagaacgata tgggaggaag
     5041 agaagaaggg tttgattggc taatcatgac cccaggtgta ggtttagacg acaaaggcga
     5101 tgcattgggt cagcagtaca gaaccgtcga cgaagttgta agtggtggat cagatatcat
     5161 cattgttggc agaggacttt tcgccaaggg tagagatcct aaggttgaag gtgaaagata
     5221 cagaaatgct ggatgggaag cgtaccaaaa gagaatcagc gctccccatt aattatacag
     5281 gaaacttaat agaacaaatc acatatttaa tctaatagcc acctgcattg gcacggtgca
     5341 acactacttc aacttcatct tacaaaaaga tcacgtgatc tgttgtattg ggatctctag
     5401 acctaataac ttcgtatagc atacattata cgaagttata ttaagggtt
//
