LOCUS       sequence-370798-        5501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Testing the SkGAL2 promoter inserted with 2 PZ4 element using a 
            green fluorescent protein as the reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5501)
  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 5501)
  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 (B11)
FEATURES             Location/Qualifiers
     source          1..5501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             772..1482
                     /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(1727..1743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1727..1743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1740..1762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1751..1767
                     /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(1775..1781,1782..1799,1800..1805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1820..1841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1958..1975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2129..2717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2209..2228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2888..3679,3680..3748))
                     /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     3511..3530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3749..3853)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3921..3939
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3977..3999)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4099..4118
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4312..4334
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4326..4343
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4327..4343
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5457..5490)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ctcgagtaaa ccaattttat ttgaacttgc cccgctaatc gtacagacgt accagatcca
       61 ggtcataacg gtgccattag aaggccgtag aatagaccca ctaaagctat cacaggcaac
      121 cttgcatcca agtctaatta aacggagcag aatctgttcc agtaatgtat aaagaattaa
      181 ctaccggaaa agcgtcttcc ggggcggctc aaccggatta cattcttttt tcttttttgc
      241 ggccagcatt cgagtcacgt gctcaaatac tgaggaaggg gttcttccgg taggcaccgg
      301 cgctcactct tccgtgcggc acactccccg cagttcaggg gccatgtgcc tgtgagagta
      361 tttccgcagg ccatagttcc ggggtggacc actccgagct aatgctaaag taagcccttc
      421 ccaacttgaa ataggaacaa aagtcaatta tctacctgaa tctaagattg ggccctagag
      481 tgagacgttg ccggcaatcc tcgagcagct cctcctgtaa agctaaacgt gtaaagattt
      541 tagagtgaga cgttgttagc agtcgttgtt caagggaggg atgagttgag gaataaaatg
      601 cggaaaaagt gtgggttaga cagacatata taaactctag ccattcggct gaaatatatc
      661 tgagttcatt tgtgataact tttttgatta cttgttcata tatccacaag aacaacgact
      721 attaaacaat agaaaatagt ataataaaaa aagtttttta caatgggatc ctctaaaggt
      781 gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt
      841 cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta
      901 aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt cactactttc
      961 ggttatggtg ttcaatgttt tgcgagatac ccagatcata tgaaacaaca tgactttttc
     1021 aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt
     1081 aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa
     1141 ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac
     1201 tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat caaagttaac
     1261 ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca ttatcaacaa
     1321 aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa
     1381 tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt
     1441 actgctgctg gtattaccca tggtatggat gaattgtaca aaagatctta aactagtagg
     1501 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
     1561 ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc
     1621 atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgggaa
     1681 tctcggtcgt aatgatttaa atgcatgcaa gcttggcgta atcatggtca tagctgtttc
     1741 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1801 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1861 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1921 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1981 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2041 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2101 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2701 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2761 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2821 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2881 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2941 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3001 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3061 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3121 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3181 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3241 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3301 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3361 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3421 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3481 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3541 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3601 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3661 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3721 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3781 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3841 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3901 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     3961 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4021 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4081 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     4141 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     4201 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     4261 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     4321 gacgttgtaa aacgacggcc agtgaattca tttaaatttt tttgattcgg taatctccga
     4381 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
     4441 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacctg
     4501 caggaaacga agataaaaca tgtccacaaa atcatatacc agtagagctg agactcatgc
     4561 aagtccggtt gcatcgaaac ttttacgttt aatggatgaa aagaagacca atttgtgtgc
     4621 ttctcttgac gttcgttcga ctgatgagct attgaaactt gttgaaacgt tgggtccata
     4681 catttgcctt ttgaaaacac acgttgatat cttggatgat ttcagttatg agggtactgt
     4741 cgttccattg aaagcattgg cagagaaata caagttcttg atatttgagg acagaaaatt
     4801 cgccgatatc ggtaacacag tcaaattaca atatacatcg ggcgtttacc gtatcgcaga
     4861 atggtctgat atcaccaacg cccacggggt tactggtgct ggtattgttg ctggcttgaa
     4921 acaaggtgcg caagaggtca ccaaagaacc aaggggatta ttgatgcttg ctgaattgtc
     4981 ttccaagggt tctctagcac acggtgaata tactaagggt accgttgata ttgcaaagag
     5041 tgataaagat ttcgttattg ggttcattgc tcagaacgat atgggaggaa gagaagaagg
     5101 gtttgattgg ctaatcatga ccccaggtgt aggtttagac gacaaaggcg atgcattggg
     5161 tcagcagtac agaaccgtcg acgaagttgt aagtggtgga tcagatatca tcattgttgg
     5221 cagaggactt ttcgccaagg gtagagatcc taaggttgaa ggtgaaagat acagaaatgc
     5281 tggatgggaa gcgtaccaaa agagaatcag cgctccccat taattataca ggaaacttaa
     5341 tagaacaaat cacatattta atctaatagc cacctgcatt ggcacggtgc aacactactt
     5401 caacttcatc ttacaaaaag atcacgtgat ctgttgtatt gggatctcta gacctaataa
     5461 cttcgtatag catacattat acgaagttat attaagggtt g
//
