LOCUS       sequence-370794-        5452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the SkGAL2 promoter inserted with 3 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 5452)
  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 5452)
  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: pILGFP10E1B
FEATURES             Location/Qualifiers
     source          1..5452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             734..1444
                     /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(1689..1705)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1689..1705)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1702..1724)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1713..1729
                     /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(1737..1743,1744..1761,1762..1767))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1782..1803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1920..1937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2091..2679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2171..2190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2850..3641,3642..3710))
                     /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     3473..3492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3711..3815)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3883..3901
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3939..3961)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4061..4080
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4274..4296
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4288..4305
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4289..4305
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5419..5452)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 attaagggtt gctcgagtaa accaatttta tttgaacttg ccccgctaat cgtacagacg
       61 taccagatcc aggtcataac ggtgccatta gaaggccgta gaatagaccc actaaagcta
      121 tcacaggcaa ccttgcatcc aagtctaatt aaacggagca gaatctgttc cagtaatgta
      181 taaagaatta actaccggaa aagcgtcttc cggggcggct caaccggatt acattctttt
      241 ttcttttttg cggccagcat tcgagtcacg tgctcaaata ctgaggaagg ggttcttccg
      301 gtaggcaccg gcgctcactc ttccgtgcgg cacactcccc gcagttcagg ggccatgtgc
      361 ctgtgagagt atttccgcag gccatagttc cggggtggac cactccgagc taatgctaaa
      421 gtaagccctt cccaacttga aataggaaca aaagtcaatt atgctcctcc tgtaaagcta
      481 aactagagtg agacgttttt agagtgagac gttgttagag tgagacgttg ttcaagggag
      541 ggatgagttg aggaataaaa tgcggaaaaa gtgtgggtta gacagacata tataaactct
      601 agccattcgg ctgaaatata tctgagttca tttgtgataa cttttttgat tacttgttca
      661 tatatccaca agaacaacga ctattaaaca atagaaaata gtataataaa aaaagttttt
      721 tacaatggga tcctctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
      781 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
      841 tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg
      901 gccaacctta gtcactactt tcggttatgg tgttcaatgt tttgcgagat acccagatca
      961 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     1021 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     1081 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     1141 aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcatgg ctgacaaaca
     1201 aaagaatggt atcaaagtta acttcaaaat tagacacaac attgaagatg gttctgttca
     1261 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     1321 caaccattac ttatccactc aatctgcctt atccaaagat ccaaacgaaa agagagacca
     1381 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     1441 caaaagatct taaactagta ggactatttg caaagggaag ggatgctaag gtagagggtg
     1501 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     1561 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     1621 tatatcagtt attacccggg aatctcggtc gtaatgattt aaatgcatgc aagcttggcg
     1681 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1741 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1801 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1861 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1921 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1981 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2041 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2101 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2161 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2221 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2281 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2341 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2401 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2461 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2521 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2581 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2641 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2701 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2761 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2821 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2881 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2941 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3001 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3061 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3121 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3181 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3241 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3301 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3361 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3421 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3481 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3541 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3601 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3661 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3721 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3781 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3841 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3901 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3961 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4021 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4081 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4141 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     4201 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     4261 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt catttaaatt
     4321 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     4381 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     4441 aactgcacag aacaaaaacc tgcaggaaac gaagataaaa catgtccaca aaatcatata
     4501 ccagtagagc tgagactcat gcaagtccgg ttgcatcgaa acttttacgt ttaatggatg
     4561 aaaagaagac caatttgtgt gcttctcttg acgttcgttc gactgatgag ctattgaaac
     4621 ttgttgaaac gttgggtcca tacatttgcc ttttgaaaac acacgttgat atcttggatg
     4681 atttcagtta tgagggtact gtcgttccat tgaaagcatt ggcagagaaa tacaagttct
     4741 tgatatttga ggacagaaaa ttcgccgata tcggtaacac agtcaaatta caatatacat
     4801 cgggcgttta ccgtatcgca gaatggtctg atatcaccaa cgcccacggg gttactggtg
     4861 ctggtattgt tgctggcttg aaacaaggtg cgcaagaggt caccaaagaa ccaaggggat
     4921 tattgatgct tgctgaattg tcttccaagg gttctctagc acacggtgaa tatactaagg
     4981 gtaccgttga tattgcaaag agtgataaag atttcgttat tgggttcatt gctcagaacg
     5041 atatgggagg aagagaagaa gggtttgatt ggctaatcat gaccccaggt gtaggtttag
     5101 acgacaaagg cgatgcattg ggtcagcagt acagaaccgt cgacgaagtt gtaagtggtg
     5161 gatcagatat catcattgtt ggcagaggac ttttcgccaa gggtagagat cctaaggttg
     5221 aaggtgaaag atacagaaat gctggatggg aagcgtacca aaagagaatc agcgctcccc
     5281 attaattata caggaaactt aatagaacaa atcacatatt taatctaata gccacctgca
     5341 ttggcacggt gcaacactac ttcaacttca tcttacaaaa agatcacgtg atctgttgta
     5401 ttgggatctc tagacctaat aacttcgtat agcatacatt atacgaagtt at
//
