LOCUS       sequence-201461-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces kudriavzevii
            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: pILGFP4I
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..717
                     /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(962..978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(962..978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(975..997)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    986..1002
                     /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(1010..1016,1017..1034,1035..1040))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1055..1076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1193..1210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1364..1952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1444..1463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2123..2914,2915..2983))
                     /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     2746..2765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2984..3088)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3156..3174
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3212..3234)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3334..3353
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3547..3569
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3561..3578
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3562..3578
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4692..4725)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ggatcctcta aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat
       61 ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac
      121 ggtaaattga ccttaaaatt tatttgtact actggtaaat tgccagttcc atggccaacc
      181 ttagtcacta ctttcggtta tggtgttcaa tgttttgcga gatacccaga tcatatgaaa
      241 caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt
      301 ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta
      361 gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac
      421 aaattggaat acaactataa ctctcacaat gtttacatca tggctgacaa acaaaagaat
      481 ggtatcaaag ttaacttcaa aattagacac aacattgaag atggttctgt tcaattagct
      541 gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat
      601 tacttatcca ctcaatctgc cttatccaaa gatccaaacg aaaagagaga ccacatggtc
      661 ttgttagaat ttgttactgc tgctggtatt acccatggta tggatgaatt gtacaaaaga
      721 tcttaaacta gtaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta
      781 cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt
      841 attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca
      901 gttattaccc gggaatctcg gtcgtaatga tttaaatgca tgcaagcttg gcgtaatcat
      961 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1021 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1081 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1141 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1201 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1261 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1321 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1801 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1921 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1981 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2041 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2101 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2161 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2221 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2281 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2341 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2401 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2461 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2521 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2581 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2641 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2701 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2761 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2821 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2881 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2941 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3001 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3061 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     3121 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     3181 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     3241 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     3301 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     3361 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3421 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     3481 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     3541 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcatttaa atttttttga
     3601 ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata
     3661 tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca
     3721 cagaacaaaa acctgcagga aacgaagata aaacatgtcc acaaaatcat ataccagtag
     3781 agctgagact catgcaagtc cggttgcatc gaaactttta cgtttaatgg atgaaaagaa
     3841 gaccaatttg tgtgcttctc ttgacgttcg ttcgactgat gagctattga aacttgttga
     3901 aacgttgggt ccatacattt gccttttgaa aacacacgtt gatatcttgg atgatttcag
     3961 ttatgagggt actgtcgttc cattgaaagc attggcagag aaatacaagt tcttgatatt
     4021 tgaggacaga aaattcgccg atatcggtaa cacagtcaaa ttacaatata catcgggcgt
     4081 ttaccgtatc gcagaatggt ctgatatcac caacgcccac ggggttactg gtgctggtat
     4141 tgttgctggc ttgaaacaag gtgcgcaaga ggtcaccaaa gaaccaaggg gattattgat
     4201 gcttgctgaa ttgtcttcca agggttctct agcacacggt gaatatacta agggtaccgt
     4261 tgatattgca aagagtgata aagatttcgt tattgggttc attgctcaga acgatatggg
     4321 aggaagagaa gaagggtttg attggctaat catgacccca ggtgtaggtt tagacgacaa
     4381 aggcgatgca ttgggtcagc agtacagaac cgtcgacgaa gttgtaagtg gtggatcaga
     4441 tatcatcatt gttggcagag gacttttcgc caagggtaga gatcctaagg ttgaaggtga
     4501 aagatacaga aatgctggat gggaagcgta ccaaaagaga atcagcgctc cccattaatt
     4561 atacaggaaa cttaatagaa caaatcacat atttaatcta atagccacct gcattggcac
     4621 ggtgcaacac tacttcaact tcatcttaca aaaagatcac gtgatctgtt gtattgggat
     4681 ctctagacct aataacttcg tatagcatac attatacgaa gttatattaa gggttgctcg
     4741 agatgtttta tttgtagagt tcaatttttc tctcttactt caaaaatgta gtgagctgta
     4801 tcgtcacact tttacggagc gctattatca taagcacgga tatttctagt cttacttata
     4861 tgttagagaa attaccaata ccccttagta ccttaaccaa aagagaaagg acctttcaat
     4921 aaggctaaaa ctcaacgctt agctaccaat ttctttgcaa tgttgaagaa cggatcagaa
     4981 actgccgagc ggacgacagc tctccgacgg gatgctctac tccgcgcgcc attgtcttca
     5041 ttagttgtgt ccctgaaaac aaacatatct cgcaccgcca tgctccgaac aataaagagt
     5101 gaaagagttg agggcaaact cccctggttt cacgtcccgt gtaaccaatg gaacaagttc
     5161 attcgcatac gatggtatcg aaattagctt tgacatgctt ctatgatggt tctgtacaag
     5221 tgggtacttt ccttccagca aagatataaa tgctaaagta aaattgatat agttgcactg
     5281 atttttcatt ccgttcactt ttttagttgc atagtttctg attttaggtt gctttaagat
     5341 aaagaaaata gcaaacaaca aatatttgat atctgtttat acattactca taaaagataa
     5401 aaataaaaat ctataatg
//
