LOCUS       sequence-201474-        5445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces paradoxus 
            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: pILGFP4O
FEATURES             Location/Qualifiers
     source          1..5445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             17..727
                     /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(972..988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(972..988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(985..1007)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    996..1012
                     /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(1020..1026,1027..1044,1045..1050))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1065..1086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1203..1220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1374..1962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1454..1473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2133..2924,2925..2993))
                     /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     2756..2775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2994..3098)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3166..3184
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3222..3244)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3344..3363
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3557..3579
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3571..3588
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3572..3588
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4702..4735)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     RBS             4756..4764
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 tttcataatg ggatcctcta aaggtgaaga attattcact ggtgttgtcc caattttggt
       61 tgaattagat ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga
      121 tgctacttac ggtaaattga ccttaaaatt tatttgtact actggtaaat tgccagttcc
      181 atggccaacc ttagtcacta ctttcggtta tggtgttcaa tgttttgcga gatacccaga
      241 tcatatgaaa caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag
      301 aactattttt ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg
      361 tgatacctta gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat
      421 tttaggtcac aaattggaat acaactataa ctctcacaat gtttacatca tggctgacaa
      481 acaaaagaat ggtatcaaag ttaacttcaa aattagacac aacattgaag atggttctgt
      541 tcaattagct gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc
      601 agacaaccat tacttatcca ctcaatctgc cttatccaaa gatccaaacg aaaagagaga
      661 ccacatggtc ttgttagaat ttgttactgc tgctggtatt acccatggta tggatgaatt
      721 gtacaaaaga tcttaaacta gtaggactat ttgcaaaggg aagggatgct aaggtagagg
      781 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
      841 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
      901 aattatatca gttattaccc gggaatctcg gtcgtaatga tttaaatgca tgcaagcttg
      961 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1021 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1081 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1141 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2701 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2761 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2821 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2881 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2941 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3001 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3061 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3121 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3181 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     3241 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     3301 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     3361 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3421 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     3481 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     3541 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcatttaa
     3601 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     3661 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     3721 cccaactgca cagaacaaaa acctgcagga aacgaagata aaacatgtcc acaaaatcat
     3781 ataccagtag agctgagact catgcaagtc cggttgcatc gaaactttta cgtttaatgg
     3841 atgaaaagaa gaccaatttg tgtgcttctc ttgacgttcg ttcgactgat gagctattga
     3901 aacttgttga aacgttgggt ccatacattt gccttttgaa aacacacgtt gatatcttgg
     3961 atgatttcag ttatgagggt actgtcgttc cattgaaagc attggcagag aaatacaagt
     4021 tcttgatatt tgaggacaga aaattcgccg atatcggtaa cacagtcaaa ttacaatata
     4081 catcgggcgt ttaccgtatc gcagaatggt ctgatatcac caacgcccac ggggttactg
     4141 gtgctggtat tgttgctggc ttgaaacaag gtgcgcaaga ggtcaccaaa gaaccaaggg
     4201 gattattgat gcttgctgaa ttgtcttcca agggttctct agcacacggt gaatatacta
     4261 agggtaccgt tgatattgca aagagtgata aagatttcgt tattgggttc attgctcaga
     4321 acgatatggg aggaagagaa gaagggtttg attggctaat catgacccca ggtgtaggtt
     4381 tagacgacaa aggcgatgca ttgggtcagc agtacagaac cgtcgacgaa gttgtaagtg
     4441 gtggatcaga tatcatcatt gttggcagag gacttttcgc caagggtaga gatcctaagg
     4501 ttgaaggtga aagatacaga aatgctggat gggaagcgta ccaaaagaga atcagcgctc
     4561 cccattaatt atacaggaaa cttaatagaa caaatcacat atttaatcta atagccacct
     4621 gcattggcac ggtgcaacac tacttcaact tcatcttaca aaaagatcac gtgatctgtt
     4681 gtattgggat ctctagacct aataacttcg tatagcatac attatacgaa gttatattaa
     4741 gggttgctcg acatttaagg aggttttacg gaccagagga gctttccaga tccagatcac
     4801 cggaatatag ggcaagagaa catcaagcag cagccgcact cgagttatca ctggcaacta
     4861 ctttgcatca aactccaatt atatgcagca gaatcttttc ttttaaaacg cacttagcgt
     4921 caactcggag ggcttccttc cgggatgact taaccagatt gcaatttctt ttccatttta
     4981 cagttaaaaa taggtcacgt gatctaaaat cgagaaaggg ttgcctcatg aaggcaccgg
     5041 ccgtcactca tccgtgcgga gaggtctgcg ccattcaggg gtccatgtgc cttagagagt
     5101 attgatgcag agggcatcgt cggggcggac cactccgaac cgacattggt taagcccttc
     5161 ccatctcaag ataggaacaa aattgcatta tactcctact agaaagctaa atgtgtacat
     5221 gttctcatga atatacaacg ttctaggtag ctattgttga gaggagggat actcagtagg
     5281 ctaaaatgtg gggaaaggat gagctttgta gacatatata aacaattaac aattggacta
     5341 aaagtttggt gttgagaatc gctcttcgct atacatcttg ttcacttatc attgagatta
     5401 accaacatta agtaaaataa agattaacat aatataaaat aattt
//
