LOCUS       sequence-201463-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces paradoxus 
            GAL1 promoter with yEGFP used as the reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  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 5418)
  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: pILGFP4K
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..720
                     /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(965..981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(965..981)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(978..1000)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    989..1005
                     /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(1013..1019,1020..1037,1038..1043))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1058..1079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1196..1213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1367..1955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1447..1466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2126..2917,2918..2986))
                     /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     2749..2768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2987..3091)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3159..3177
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3215..3237)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3337..3356
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3550..3572
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3564..3581
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3565..3581
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4695..4728)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 atgggatcct ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta
       61 gatggtgatg ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact
      121 tacggtaaat tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca
      181 accttagtca ctactttcgg ttatggtgtt caatgttttg cgagataccc agatcatatg
      241 aaacaacatg actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt
      301 tttttcaaag atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc
      361 ttagttaata gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt
      421 cacaaattgg aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag
      481 aatggtatca aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta
      541 gctgaccatt atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac
      601 cattacttat ccactcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg
      661 gtcttgttag aatttgttac tgctgctggt attacccatg gtatggatga attgtacaaa
      721 agatcttaaa ctagtaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
      781 ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
      841 tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
      901 tcagttatta cccgggaatc tcggtcgtaa tgatttaaat gcatgcaagc ttggcgtaat
      961 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1021 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1081 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1141 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1201 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1801 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2041 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2101 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2161 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2221 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2281 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2341 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2401 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2461 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2521 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2581 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2641 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2701 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2761 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2821 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2881 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2941 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3001 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3061 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3121 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3181 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3241 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3301 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     3361 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3421 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     3481 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     3541 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcatt taaatttttt
     3601 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     3661 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     3721 gcacagaaca aaaacctgca ggaaacgaag ataaaacatg tccacaaaat catataccag
     3781 tagagctgag actcatgcaa gtccggttgc atcgaaactt ttacgtttaa tggatgaaaa
     3841 gaagaccaat ttgtgtgctt ctcttgacgt tcgttcgact gatgagctat tgaaacttgt
     3901 tgaaacgttg ggtccataca tttgcctttt gaaaacacac gttgatatct tggatgattt
     3961 cagttatgag ggtactgtcg ttccattgaa agcattggca gagaaataca agttcttgat
     4021 atttgaggac agaaaattcg ccgatatcgg taacacagtc aaattacaat atacatcggg
     4081 cgtttaccgt atcgcagaat ggtctgatat caccaacgcc cacggggtta ctggtgctgg
     4141 tattgttgct ggcttgaaac aaggtgcgca agaggtcacc aaagaaccaa ggggattatt
     4201 gatgcttgct gaattgtctt ccaagggttc tctagcacac ggtgaatata ctaagggtac
     4261 cgttgatatt gcaaagagtg ataaagattt cgttattggg ttcattgctc agaacgatat
     4321 gggaggaaga gaagaagggt ttgattggct aatcatgacc ccaggtgtag gtttagacga
     4381 caaaggcgat gcattgggtc agcagtacag aaccgtcgac gaagttgtaa gtggtggatc
     4441 agatatcatc attgttggca gaggactttt cgccaagggt agagatccta aggttgaagg
     4501 tgaaagatac agaaatgctg gatgggaagc gtaccaaaag agaatcagcg ctccccatta
     4561 attatacagg aaacttaata gaacaaatca catatttaat ctaatagcca cctgcattgg
     4621 cacggtgcaa cactacttca acttcatctt acaaaaagat cacgtgatct gttgtattgg
     4681 gatctctaga cctaataact tcgtatagca tacattatac gaagttatat taagggttgc
     4741 tcgagctatg ttgatctttt cagaattttt cactatatta agatgggtgc aaagaagtgt
     4801 gattattata ttacatcgct ttcctatcat acacatatcc atatctagtc ttacttatat
     4861 gttgtgagag tgttgataac cccagtatct taacccaaga aagcctttct atgaaacttg
     4921 aactgtacgc taaactgctc attgcaatat tgaagtacgg atcagaagcc gccgagcgga
     4981 cgacagccct ccgacggaat attcccctcc gtgcgtcgcc gtcttcgtcg gttgtgtccc
     5041 ttaacatcga tgtacctcgc gccgccctgc tccgaacaat aaggattcta caagaaatac
     5101 ttgttttttt atggttatga caggaacaaa ataagcagcc cactgacccc atataccttt
     5161 caaactattg aatcaaattg gccagcatat ggtaatagta cagttagggt tttccttatt
     5221 cctgagacaa ttcatccgca aaaaataatg gtttttggtc tattagcaaa catataaatg
     5281 caaaagttgc atagccactt taaatacatt tgctcctcca agatttttaa tttcgttttg
     5341 ttttattgtc atggaaatat taacaacaag tagttaatat acatctatac tttatcgtca
     5401 aggaaagaac aaactata
//
