LOCUS       sequence-201470-        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             15..725
                     /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(970..986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(970..986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(983..1005)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    994..1010
                     /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(1018..1024,1025..1042,1043..1048))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1063..1084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1201..1218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1372..1960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1452..1471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2131..2922,2923..2991))
                     /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     2754..2773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2992..3096)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3164..3182
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3220..3242)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3342..3361
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3555..3577
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3569..3586
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3570..3586
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4700..4733)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ttacaatggg atcctctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg
       61 aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg
      121 ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat
      181 ggccaacctt agtcactact ttcggttatg gtgttcaatg ttttgcgaga tacccagatc
      241 atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa
      301 ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg
      361 ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt
      421 taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg gctgacaaac
      481 aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggttctgttc
      541 aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag
      601 acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa aagagagacc
      661 acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattgt
      721 acaaaagatc ttaaactagt aggactattt gcaaagggaa gggatgctaa ggtagagggt
      781 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
      841 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
      901 ttatatcagt tattacccgg gaatctcggt cgtaatgatt taaatgcatg caagcttggc
      961 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1021 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1081 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1141 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1201 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1261 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1321 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1381 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1441 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1501 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1561 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1621 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1681 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1741 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1801 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1861 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1921 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1981 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2041 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2161 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2221 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2281 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2341 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2401 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2461 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2521 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2581 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2641 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2701 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2761 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2821 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2881 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2941 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3061 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3121 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3181 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     3241 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     3301 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     3361 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     3421 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3481 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     3541 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcatttaaat
     3601 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     3661 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     3721 caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa acatgtccac aaaatcatat
     3781 accagtagag ctgagactca tgcaagtccg gttgcatcga aacttttacg tttaatggat
     3841 gaaaagaaga ccaatttgtg tgcttctctt gacgttcgtt cgactgatga gctattgaaa
     3901 cttgttgaaa cgttgggtcc atacatttgc cttttgaaaa cacacgttga tatcttggat
     3961 gatttcagtt atgagggtac tgtcgttcca ttgaaagcat tggcagagaa atacaagttc
     4021 ttgatatttg aggacagaaa attcgccgat atcggtaaca cagtcaaatt acaatataca
     4081 tcgggcgttt accgtatcgc agaatggtct gatatcacca acgcccacgg ggttactggt
     4141 gctggtattg ttgctggctt gaaacaaggt gcgcaagagg tcaccaaaga accaagggga
     4201 ttattgatgc ttgctgaatt gtcttccaag ggttctctag cacacggtga atatactaag
     4261 ggtaccgttg atattgcaaa gagtgataaa gatttcgtta ttgggttcat tgctcagaac
     4321 gatatgggag gaagagaaga agggtttgat tggctaatca tgaccccagg tgtaggttta
     4381 gacgacaaag gcgatgcatt gggtcagcag tacagaaccg tcgacgaagt tgtaagtggt
     4441 ggatcagata tcatcattgt tggcagagga cttttcgcca agggtagaga tcctaaggtt
     4501 gaaggtgaaa gatacagaaa tgctggatgg gaagcgtacc aaaagagaat cagcgctccc
     4561 cattaattat acaggaaact taatagaaca aatcacatat ttaatctaat agccacctgc
     4621 attggcacgg tgcaacacta cttcaacttc atcttacaaa aagatcacgt gatctgttgt
     4681 attgggatct ctagacctaa taacttcgta tagcatacat tatacgaagt tatattaagg
     4741 gttgctcgac ccacagagaa caggagatta caagatccag atcaccaaaa tgccggaaca
     4801 caggctgaaa aactccaagc agccgcacac ccgattttta gccagcagat tcatacctaa
     4861 ctccaatgaa ttgcagcata tatcctgcaa taacacatta aggatcgggc cttggacggc
     4921 cgaatatatg attatagtcc ttttgtcgcc aaaaagtcgc ctcgaggtca cgtgcatccg
     4981 aaatcacccc ctgagcactt cccgcccaga aaacagggaa ggggcggcac cagcaggcac
     5041 cggcattcac tcgtccgtgc ggcgtggtct gtgccgttca ggggcctgcg tgcctgtgat
     5101 gatatcgctg cagggcaccg agtcggagag gaccactccg agccgaggtt agttaagcca
     5161 ttctctacgc taaaaaataa ataatgtgga ttaagctcct actggaaggt taagtgtttg
     5221 tacactttca aacatgcgca acgcaccagg caactattgt tctgaggagg gatgtgatgc
     5281 ggagtaaaat gtggaggaat tacggggctt gcagacatat ataaactcta gccattcggc
     5341 tgaatataag ctagaaacca cttcttctct ttccatctac atctccctca tctatcaata
     5401 agaactataa tatcaacaat actataatat aatatataca cacat
//
