LOCUS       sequence-370796-        5566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the SkGAL2 promoter inserted with 4 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 5566)
  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 5566)
  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: pILGFP10CF72
FEATURES             Location/Qualifiers
     source          1..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             842..1552
                     /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(1797..1813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1797..1813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1810..1832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1821..1837
                     /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(1845..1851,1852..1869,1870..1875))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1890..1911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2028..2045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2199..2787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2279..2298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2958..3749,3750..3818))
                     /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     3581..3600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3819..3923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3991..4009
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4047..4069)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4169..4188
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4382..4404
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4396..4413
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4397..4413
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5527..5560)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ggttgctcga gtaaaccaat tttatttgaa cttgccccgc taatcgtaca gacgtaccag
       61 atccaggtca taacggtgcc attagaaggc cgtagaatag acccactaaa gctatcacag
      121 gcaaccttgc atccaagtct aattaaacgg agcagaatct gttccagtaa tgtataaaga
      181 attaactacc ggaaaagcgt cttccggggc ggctcaaccg gattacattc ttttttcttt
      241 tttgcggcca gcattcgagt cacgtgctca aatactgagg aaggggttct tccggtaggc
      301 accggcgctc actcttccgt gcggcacact ccccgcagtt caggggccat gtgcctgtga
      361 gagtatttcc gcaggccata gttccggggt ggaccactcc gagctaatgc taaagtaagc
      421 ccttcccaac ttgaaatagg aacaaaagtc aattatctac ctgaatctaa gattgggccc
      481 tagagtgaga cgttaattgg tagagtgaga cgtttaagcg ggggcggtag agtgagacgt
      541 tgggcttaga gtgaacgttg ccggcaatcc tcgagcagct cctcctgtaa agctaaacgt
      601 gtaaagattt tagagtgaga cgttgttagc agtcgttgtt caagggaggg atgagttgag
      661 gaataaaatg cggaaaaagt gtgggttaga cagacatata taaactctag ccattcggct
      721 gaaatatatc tgagttcatt tgtgataact tttttgatta cttgttcata tatccacaag
      781 aacaacgact attaaacaat agaaaatagt ataataaaaa aagtttttta caatgggatc
      841 ctctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
      901 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
      961 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
     1021 cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata tgaaacaaca
     1081 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
     1141 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
     1201 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
     1261 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
     1321 caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca
     1381 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
     1441 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
     1501 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aaagatctta
     1561 aactagtagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     1621 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     1681 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     1741 tacccgggaa tctcggtcgt aatgatttaa atgcatgcaa gcttggcgta atcatggtca
     1801 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1861 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1921 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1981 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2041 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2101 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2161 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2221 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2281 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2341 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2401 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2461 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2521 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2581 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2641 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2701 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2761 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2821 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2881 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2941 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3001 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3061 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3121 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3181 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3241 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3301 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3361 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3421 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3481 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3541 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3601 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3661 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3721 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3781 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3841 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3901 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     3961 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4021 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4081 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4141 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4201 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     4261 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4321 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     4381 tcccagtcac gacgttgtaa aacgacggcc agtgaattca tttaaatttt tttgattcgg
     4441 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     4501 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     4561 caaaaacctg caggaaacga agataaaaca tgtccacaaa atcatatacc agtagagctg
     4621 agactcatgc aagtccggtt gcatcgaaac ttttacgttt aatggatgaa aagaagacca
     4681 atttgtgtgc ttctcttgac gttcgttcga ctgatgagct attgaaactt gttgaaacgt
     4741 tgggtccata catttgcctt ttgaaaacac acgttgatat cttggatgat ttcagttatg
     4801 agggtactgt cgttccattg aaagcattgg cagagaaata caagttcttg atatttgagg
     4861 acagaaaatt cgccgatatc ggtaacacag tcaaattaca atatacatcg ggcgtttacc
     4921 gtatcgcaga atggtctgat atcaccaacg cccacggggt tactggtgct ggtattgttg
     4981 ctggcttgaa acaaggtgcg caagaggtca ccaaagaacc aaggggatta ttgatgcttg
     5041 ctgaattgtc ttccaagggt tctctagcac acggtgaata tactaagggt accgttgata
     5101 ttgcaaagag tgataaagat ttcgttattg ggttcattgc tcagaacgat atgggaggaa
     5161 gagaagaagg gtttgattgg ctaatcatga ccccaggtgt aggtttagac gacaaaggcg
     5221 atgcattggg tcagcagtac agaaccgtcg acgaagttgt aagtggtgga tcagatatca
     5281 tcattgttgg cagaggactt ttcgccaagg gtagagatcc taaggttgaa ggtgaaagat
     5341 acagaaatgc tggatgggaa gcgtaccaaa agagaatcag cgctccccat taattataca
     5401 ggaaacttaa tagaacaaat cacatattta atctaatagc cacctgcatt ggcacggtgc
     5461 aacactactt caacttcatc ttacaaaaag atcacgtgat ctgttgtatt gggatctcta
     5521 gacctaataa cttcgtatag catacattat acgaagttat attaag
//
