LOCUS       sequence-201466-        5445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces mikatae 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: pILGFP4L
FEATURES             Location/Qualifiers
     source          1..5445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..722
                     /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(967..983)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(967..983)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(980..1002)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    991..1007
                     /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(1015..1021,1022..1039,1040..1045))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1060..1081
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1198..1215)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1369..1957)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1449..1468)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2128..2919,2920..2988))
                     /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     2751..2770
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2989..3093)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3161..3179
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3217..3239)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3339..3358
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3552..3574
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3566..3583
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3567..3583
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4697..4730)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 taatgggatc ctctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat
       61 tagatggtga tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta
      121 cttacggtaa attgacctta aaatttattt gtactactgg taaattgcca gttccatggc
      181 caaccttagt cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata
      241 tgaaacaaca tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta
      301 tttttttcaa agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata
      361 ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag
      421 gtcacaaatt ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa
      481 agaatggtat caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat
      541 tagctgacca ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca
      601 accattactt atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca
      661 tggtcttgtt agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca
      721 aaagatctta aactagtagg actatttgca aagggaaggg atgctaaggt agagggtgaa
      781 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
      841 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
      901 tatcagttat tacccgggaa tctcggtcgt aatgatttaa atgcatgcaa gcttggcgta
      961 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1021 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1201 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1261 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1321 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1381 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1441 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1501 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1561 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1621 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1681 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1741 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1801 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1861 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1921 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1981 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2041 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2101 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2161 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2221 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2281 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2341 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2401 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2461 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2521 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2581 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2641 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2701 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2761 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2821 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2881 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2941 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3001 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3061 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3121 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     3181 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3241 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3301 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     3361 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     3421 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     3481 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     3541 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattca tttaaatttt
     3601 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     3661 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     3721 ctgcacagaa caaaaacctg caggaaacga agataaaaca tgtccacaaa atcatatacc
     3781 agtagagctg agactcatgc aagtccggtt gcatcgaaac ttttacgttt aatggatgaa
     3841 aagaagacca atttgtgtgc ttctcttgac gttcgttcga ctgatgagct attgaaactt
     3901 gttgaaacgt tgggtccata catttgcctt ttgaaaacac acgttgatat cttggatgat
     3961 ttcagttatg agggtactgt cgttccattg aaagcattgg cagagaaata caagttcttg
     4021 atatttgagg acagaaaatt cgccgatatc ggtaacacag tcaaattaca atatacatcg
     4081 ggcgtttacc gtatcgcaga atggtctgat atcaccaacg cccacggggt tactggtgct
     4141 ggtattgttg ctggcttgaa acaaggtgcg caagaggtca ccaaagaacc aaggggatta
     4201 ttgatgcttg ctgaattgtc ttccaagggt tctctagcac acggtgaata tactaagggt
     4261 accgttgata ttgcaaagag tgataaagat ttcgttattg ggttcattgc tcagaacgat
     4321 atgggaggaa gagaagaagg gtttgattgg ctaatcatga ccccaggtgt aggtttagac
     4381 gacaaaggcg atgcattggg tcagcagtac agaaccgtcg acgaagttgt aagtggtgga
     4441 tcagatatca tcattgttgg cagaggactt ttcgccaagg gtagagatcc taaggttgaa
     4501 ggtgaaagat acagaaatgc tggatgggaa gcgtaccaaa agagaatcag cgctccccat
     4561 taattataca ggaaacttaa tagaacaaat cacatattta atctaatagc cacctgcatt
     4621 ggcacggtgc aacactactt caacttcatc ttacaaaaag atcacgtgat ctgttgtatt
     4681 gggatctcta gacctaataa cttcgtatag catacattat acgaagttat attaagggtt
     4741 gctcgagtta cggatcagag gagttctcca gatctgaatc acgatgattt cggaaaacac
     4801 agggcaaaga gaaacaaggt agccgcactc aattcatctt ttagtgacta cttttcatcg
     4861 aaatccaatt aaatgcagta gaatctgttc aaaaaaatgt acatagtgtc aattcggcaa
     4921 gctttcttcc ggaatggctt aaccagacca tatatctctt cattattttc attttaaggt
     4981 ccggagtatt gatcacgtga tcaaattccc attgaagggt tgccttttaa aggcaccggt
     5041 agtcactcat ccgtgcggag tgttctgcgc cattcagggg ccaatgtgcc tttgagaaga
     5101 ttagagcagg gagcagactc ggggcggacc actccgagcc gatgttagtt aagccttccc
     5161 atcttgaaat aggaacaaaa tggctttata ctcctgccag aaagcaaggc atgtgtatac
     5221 cttctcaaga atatacaacg ttttggcagc tattgttcag aagagggata gatcgtaagc
     5281 taaaatttgg agaaattaaa gatttgtaga catatataaa ctatcagcaa ttcactgcta
     5341 aaatacatct tgagagtaac tcttcgctta ttcatcttgt tcacttttcg atgaagaaaa
     5401 ccactgttaa gtaaaataaa gaataacata taaaaaactt tttca
//
