LOCUS       sequence-201440-        5439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces cerevisiae 
            PGM2 promoter with yEGFP used as the reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5439)
  AUTHORS   Peng B, Wood RJ, Nielsen LK, Vickers CE
  TITLE     An Expanded Heterologous GAL Promoter Collection for 
            Diauxie-Inducible Expression in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):748-751. doi: 
            10.1021/acssynbio.7b00355. Epub 2018 Jan 23.
  PUBMED    29301066
REFERENCE   2  (bases 1 to 5439)
  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"; doi: 
            "10.1021/acssynbio.7b00355"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "748-751"
COMMENT     Sequence Label: pILGFP4G
FEATURES             Location/Qualifiers
     source          1..5439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             11..721
                     /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(966..982)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(966..982)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(979..1001)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    990..1006
                     /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(1014..1020,1021..1038,1039..1044))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1059..1080
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1197..1214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1368..1956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1448..1467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2127..2918,2919..2987))
                     /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     2750..2769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2988..3092)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3160..3178
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3216..3238)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3338..3357
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3551..3573
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3565..3582
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3566..3582
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4696..4729)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 catgggatcc tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt
       61 agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac
      121 ttacggtaaa ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc
      181 aaccttagtc actactttcg gttatggtgt tcaatgtttt gcgagatacc cagatcatat
      241 gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat
      301 ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac
      361 cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg
      421 tcacaaattg gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa
      481 gaatggtatc aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt
      541 agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa
      601 ccattactta tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat
      661 ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa
      721 aagatcttaa actagtagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
      781 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
      841 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
      901 atcagttatt acccgggaat ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa
      961 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1021 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1081 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1141 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1201 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1261 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1321 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1381 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1441 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1501 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1561 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1621 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1681 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1741 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1801 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1861 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1921 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1981 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2041 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2101 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2161 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2221 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2281 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2341 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2401 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2461 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2521 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2581 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2641 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2701 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2761 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2821 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2881 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2941 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3001 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3061 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3121 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3181 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3241 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3301 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     3361 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     3421 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     3481 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     3541 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcat ttaaattttt
     3601 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     3661 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     3721 tgcacagaac aaaaacctgc aggaaacgaa gataaaacat gtccacaaaa tcatatacca
     3781 gtagagctga gactcatgca agtccggttg catcgaaact tttacgttta atggatgaaa
     3841 agaagaccaa tttgtgtgct tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg
     3901 ttgaaacgtt gggtccatac atttgccttt tgaaaacaca cgttgatatc ttggatgatt
     3961 tcagttatga gggtactgtc gttccattga aagcattggc agagaaatac aagttcttga
     4021 tatttgagga cagaaaattc gccgatatcg gtaacacagt caaattacaa tatacatcgg
     4081 gcgtttaccg tatcgcagaa tggtctgata tcaccaacgc ccacggggtt actggtgctg
     4141 gtattgttgc tggcttgaaa caaggtgcgc aagaggtcac caaagaacca aggggattat
     4201 tgatgcttgc tgaattgtct tccaagggtt ctctagcaca cggtgaatat actaagggta
     4261 ccgttgatat tgcaaagagt gataaagatt tcgttattgg gttcattgct cagaacgata
     4321 tgggaggaag agaagaaggg tttgattggc taatcatgac cccaggtgta ggtttagacg
     4381 acaaaggcga tgcattgggt cagcagtaca gaaccgtcga cgaagttgta agtggtggat
     4441 cagatatcat cattgttggc agaggacttt tcgccaaggg tagagatcct aaggttgaag
     4501 gtgaaagata cagaaatgct ggatgggaag cgtaccaaaa gagaatcagc gctccccatt
     4561 aattatacag gaaacttaat agaacaaatc acatatttaa tctaatagcc acctgcattg
     4621 gcacggtgca acactacttc aacttcatct tacaaaaaga tcacgtgatc tgttgtattg
     4681 ggatctctag acctaataac ttcgtatagc atacattata cgaagttata ttaagggttg
     4741 ctcgagaaaa ggtctaacat cctttgagtt atgttttcgt caccacagtt aaagagaaaa
     4801 aaaaataaat agcgggtaaa taattgtcat actactgatt tgcattgtta ccttgtaact
     4861 ttgaacacta gccatgcagg ttgcttttcc tcctttacac cttcccccct gtcgctgttc
     4921 aaaagtaaat gcaacaacaa taaaacgtgg gtagaggacg ttccctgtat gtactaaaat
     4981 catactatca tccgttgagg tcgagcggtt ctctaatctc aacacacgct gctgcccctg
     5041 agcttgcgag ctaaggacaa cagggccaac ttgttctttc taggggggag aggcccaact
     5101 taggctcggt gaagagaagc gcggagctca cgaaaagggg tggtgtcgcg cactactttt
     5161 ttcgtgggga acttacgtga aaggggacgt tatacttttg ttatgtgcag ttccaggaac
     5221 aaacaggggt tgaaaagtgt aaaagaaata gttgattctt tcagaatctg gatgactgcc
     5281 gcccgaacaa tgtgggaaat ataaaaaggt accgttaata ttgtttcatt ttcgaagata
     5341 gtgttccatg tctttcttca gcgttgcttg aacaatagga taataagaag aagatcaacc
     5401 aatctttctc agtaaaaaaa gtaacaaaag ttaacataa
//
