LOCUS       sequence-198121-        5445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces eubayanus 
            GAL2 promoter with yEGFP used as the reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5445)
  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 5445)
  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: pILGFP4M
FEATURES             Location/Qualifiers
     source          1..5445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..716
                     /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(961..977)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(961..977)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(974..996)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    985..1001
                     /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(1009..1015,1016..1033,1034..1039))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1054..1075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1192..1209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1363..1951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1443..1462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2122..2913,2914..2982))
                     /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     2745..2764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2983..3087)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3155..3173
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3211..3233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3333..3352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3546..3568
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3560..3577
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3561..3577
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4691..4724)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gatcctctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg
       61 gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg
      121 gtaaattgac cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct
      181 tagtcactac tttcggttat ggtgttcaat gttttgcgag atacccagat catatgaaac
      241 aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt
      301 tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag
      361 ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca
      421 aattggaata caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg
      481 gtatcaaagt taacttcaaa attagacaca acattgaaga tggttctgtt caattagctg
      541 accattatca acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt
      601 acttatccac tcaatctgcc ttatccaaag atccaaacga aaagagagac cacatggtct
      661 tgttagaatt tgttactgct gctggtatta cccatggtat ggatgaattg tacaaaagat
      721 cttaaactag taggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
      781 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
      841 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
      901 ttattacccg ggaatctcgg tcgtaatgat ttaaatgcat gcaagcttgg cgtaatcatg
      961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1801 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3121 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3181 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3241 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3301 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     3361 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     3421 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3481 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3541 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcatttaaa tttttttgat
     3601 tcggtaatct ccgagcagaa ggaagaacga aggaaggagc acagacttag attggtatat
     3661 atacgcatat gtggtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     3721 agaacaaaaa cctgcaggaa acgaagataa aacatgtcca caaaatcata taccagtaga
     3781 gctgagactc atgcaagtcc ggttgcatcg aaacttttac gtttaatgga tgaaaagaag
     3841 accaatttgt gtgcttctct tgacgttcgt tcgactgatg agctattgaa acttgttgaa
     3901 acgttgggtc catacatttg ccttttgaaa acacacgttg atatcttgga tgatttcagt
     3961 tatgagggta ctgtcgttcc attgaaagca ttggcagaga aatacaagtt cttgatattt
     4021 gaggacagaa aattcgccga tatcggtaac acagtcaaat tacaatatac atcgggcgtt
     4081 taccgtatcg cagaatggtc tgatatcacc aacgcccacg gggttactgg tgctggtatt
     4141 gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg attattgatg
     4201 cttgctgaat tgtcttccaa gggttctcta gcacacggtg aatatactaa gggtaccgtt
     4261 gatattgcaa agagtgataa agatttcgtt attgggttca ttgctcagaa cgatatggga
     4321 ggaagagaag aagggtttga ttggctaatc atgaccccag gtgtaggttt agacgacaaa
     4381 ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg tggatcagat
     4441 atcatcattg ttggcagagg acttttcgcc aagggtagag atcctaaggt tgaaggtgaa
     4501 agatacagaa atgctggatg ggaagcgtac caaaagagaa tcagcgctcc ccattaatta
     4561 tacaggaaac ttaatagaac aaatcacata tttaatctaa tagccacctg cattggcacg
     4621 gtgcaacact cacttcaact tcatcttaca aaagatcacg tgatctgttg tattgggatc
     4681 tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttgctcga
     4741 cccacagaga acaggagatt acaagatcca gatcaccaaa atgccggaac acaggctgaa
     4801 aaactccaag cagccgcaca cccgattttt agccagcaga ttcataccta actccaatga
     4861 attgcagcat atatcctgca ataacacatt aaggatcggg ccttggacgg ccgaatatat
     4921 gattatagtc cttttgtcgc caaaaagtcg cctcgaggtc acgtgcatcc gaaatcaccc
     4981 cctgagcact tcccgcccag aaaacaggga aggggcggca ccagcaggca ccggcattca
     5041 ctcgtccgtg cggcgtggtc tgtgccgttc aggggcctgc gtgcctgtga tgatatcgct
     5101 gcagggcacc gagtcggaga ggaccactcc gagccgaggt tagttaagcc attctctacg
     5161 ctaaaaaata aataatgtgg attaagctcc tactggaagg ttaagtgttt gtacactttc
     5221 aaacatgcgc aacgcaccag gcaactattg ttctgaggag ggatgtgatg cggagtaaaa
     5281 tgtggaggaa ttacggggct tgcagacata tataaactct agccattcgg ctgaatataa
     5341 gctagaaacc acttcttctc tttccatcta catctccctc atctatcaat aagaactata
     5401 atatcaacaa tactataata taatatatac acacatttac aatgg
//
