LOCUS       sequence-201475-        5444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces pastorianus 
            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 5444)
  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 5444)
  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: pILGFP4P
FEATURES             Location/Qualifiers
     source          1..5444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             19..729
                     /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(974..990)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(974..990)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(987..1009)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    998..1014
                     /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(1022..1028,1029..1046,1047..1052))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1067..1088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1205..1222)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1376..1964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1456..1475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2135..2926,2927..2995))
                     /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     2758..2777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2996..3100)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3168..3186
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3224..3246)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3346..3365
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3559..3581
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3573..3590
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3574..3590
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4704..4737)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 acattcaaaa tgggatcctc taaaggtgaa gaattattca ctggtgttgt cccaattttg
       61 gttgaattag atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt
      121 gatgctactt acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt
      181 ccatggccaa ccttagtcac tactttcggt tatggtgttc aatgttttgc gagataccca
      241 gatcatatga aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa
      301 agaactattt ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa
      361 ggtgatacct tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac
      421 attttaggtc acaaattgga atacaactat aactctcaca atgtttacat catggctgac
      481 aaacaaaaga atggtatcaa agttaacttc aaaattagac acaacattga agatggttct
      541 gttcaattag ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta
      601 ccagacaacc attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga
      661 gaccacatgg tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa
      721 ttgtacaaaa gatcttaaac tagtaggact atttgcaaag ggaagggatg ctaaggtaga
      781 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
      841 ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
      901 ttaattatat cagttattac ccgggaatct cggtcgtaat gatttaaatg catgcaagct
      961 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1021 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1081 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1141 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1201 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1261 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1321 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1381 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1441 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1501 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1561 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1621 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1681 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1741 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1801 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1861 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1921 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1981 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2041 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2101 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2161 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2221 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2281 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2341 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2401 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2461 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2521 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2581 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2641 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2701 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2761 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2821 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2881 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2941 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3001 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3061 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3121 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3181 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     3241 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     3301 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     3361 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3421 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     3481 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     3541 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcattt
     3601 aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact
     3661 tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt
     3721 aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaaacatgt ccacaaaatc
     3781 atataccagt agagctgaga ctcatgcaag tccggttgca tcgaaacttt tacgtttaat
     3841 ggatgaaaag aagaccaatt tgtgtgcttc tcttgacgtt cgttcgactg atgagctatt
     3901 gaaacttgtt gaaacgttgg gtccatacat ttgccttttg aaaacacacg ttgatatctt
     3961 ggatgatttc agttatgagg gtactgtcgt tccattgaaa gcattggcag agaaatacaa
     4021 gttcttgata tttgaggaca gaaaattcgc cgatatcggt aacacagtca aattacaata
     4081 tacatcgggc gtttaccgta tcgcagaatg gtctgatatc accaacgccc acggggttac
     4141 tggtgctggt attgttgctg gcttgaaaca aggtgcgcaa gaggtcacca aagaaccaag
     4201 gggattattg atgcttgctg aattgtcttc caagggttct ctagcacacg gtgaatatac
     4261 taagggtacc gttgatattg caaagagtga taaagatttc gttattgggt tcattgctca
     4321 gaacgatatg ggaggaagag aagaagggtt tgattggcta atcatgaccc caggtgtagg
     4381 tttagacgac aaaggcgatg cattgggtca gcagtacaga accgtcgacg aagttgtaag
     4441 tggtggatca gatatcatca ttgttggcag aggacttttc gccaagggta gagatcctaa
     4501 ggttgaaggt gaaagataca gaaatgctgg atgggaagcg taccaaaaga gaatcagcgc
     4561 tccccattaa ttatacagga aacttaatag aacaaatcac atatttaatc taatagccac
     4621 ctgcattggc acggtgcaac actacttcaa cttcatctta caaaaagatc acgtgatctg
     4681 ttgtattggg atctctagac ctaataactt cgtatagcat acattatacg aagttatatt
     4741 aagggttgct cgagacccca ccagatccgg aagaccacag agacagacga ttacaagatc
     4801 cggttcacaa gaacaccgga ccacaggcta gaagggactc caatgtgccg cacatctgat
     4861 atgcagtaat taagctcgca cctagctcca ataagttgca gcatgaatat tgcaataaca
     4921 caccgagagt gttgggtctt gaacggccga ttgtagtccg tttgtcgcca taaaagtctc
     4981 cttgaggtca cgtgcaccta ggattacccc ttgagcaact ttccgcccag aaatcaagga
     5041 agagaggcac tggaaggcac cggctgccac cagtccgtgc agtgggtctg tgacattcag
     5101 ggaccagggt gcctttgaaa aaatcggtgc agggcacaaa tcggagagga ccactccgag
     5161 cagaggttag ttaagccatt ttctacgcta aaaaaaaata aggtggatta agctcctagc
     5221 ggaaagttaa gagtgtgtac gcttccagat atgcacaacg tgcgaggcaa ctattgttcc
     5281 aaagagggat gtgttgcgga gtataatgcg gaggtcatat ggggcttgca gacatatata
     5341 agctctggcc attcggctga aaataaacta ggatctatca tcgtctttcc atctatttct
     5401 cactcatcca acaatatcaa cagtacaaat aatattatat atac
//
