LOCUS       sequence-201473-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces bayanus 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 5455)
  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 5455)
  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: pILGFP4N
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             18..728
                     /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(973..989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(973..989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(986..1008)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    997..1013
                     /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(1021..1027,1028..1045,1046..1051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1066..1087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1204..1221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1375..1963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1455..1474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2134..2925,2926..2994))
                     /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     2757..2776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2995..3099)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3167..3185
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3223..3245)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3345..3364
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3558..3580
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3572..3589
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3573..3589
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4703..4736)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cattcaaaat gggatcctct aaaggtgaag aattattcac tggtgttgtc ccaattttgg
       61 ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg
      121 atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa ttgccagttc
      181 catggccaac cttagtcact actttcggtt atggtgttca atgttttgcg agatacccag
      241 atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat gttcaagaaa
      301 gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc aagtttgaag
      361 gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa gatggtaaca
      421 ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc atggctgaca
      481 aacaaaagaa tggtatcaaa gttaacttca aaattagaca caacattgaa gatggttctg
      541 ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca gtcttgttac
      601 cagacaacca ttacttatcc actcaatctg ccttatccaa agatccaaac gaaaagagag
      661 accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt atggatgaat
      721 tgtacaaaag atcttaaact agtaggacta tttgcaaagg gaagggatgc taaggtagag
      781 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
      841 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
      901 taattatatc agttattacc cgggaatctc ggtcgtaatg atttaaatgc atgcaagctt
      961 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1021 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1081 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1141 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2401 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3121 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3181 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3241 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3301 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3361 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     3421 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     3481 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     3541 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcattta
     3601 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     3661 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     3721 acccaactgc acagaacaaa aacctgcagg aaacgaagat aaaacatgtc cacaaaatca
     3781 tataccagta gagctgagac tcatgcaagt ccggttgcat cgaaactttt acgtttaatg
     3841 gatgaaaaga agaccaattt gtgtgcttct cttgacgttc gttcgactga tgagctattg
     3901 aaacttgttg aaacgttggg tccatacatt tgccttttga aaacacacgt tgatatcttg
     3961 gatgatttca gttatgaggg tactgtcgtt ccattgaaag cattggcaga gaaatacaag
     4021 ttcttgatat ttgaggacag aaaattcgcc gatatcggta acacagtcaa attacaatat
     4081 acatcgggcg tttaccgtat cgcagaatgg tctgatatca ccaacgccca cggggttact
     4141 ggtgctggta ttgttgctgg cttgaaacaa ggtgcgcaag aggtcaccaa agaaccaagg
     4201 ggattattga tgcttgctga attgtcttcc aagggttctc tagcacacgg tgaatatact
     4261 aagggtaccg ttgatattgc aaagagtgat aaagatttcg ttattgggtt cattgctcag
     4321 aacgatatgg gaggaagaga agaagggttt gattggctaa tcatgacccc aggtgtaggt
     4381 ttagacgaca aaggcgatgc attgggtcag cagtacagaa ccgtcgacga agttgtaagt
     4441 ggtggatcag atatcatcat tgttggcaga ggacttttcg ccaagggtag agatcctaag
     4501 gttgaaggtg aaagatacag aaatgctgga tgggaagcgt accaaaagag aatcagcgct
     4561 ccccattaat tatacaggaa acttaataga acaaatcaca tatttaatct aatagccacc
     4621 tgcattggca cggtgcaaca ctacttcaac ttcatcttac aaaaagatca cgtgatctgt
     4681 tgtattggga tctctagacc taataacttc gtatagcata cattatacga agttatatta
     4741 agggttgctc gacccacaac cgcaccagat ccggaagaca cagagaacag gagattacca
     4801 gatccagatc agcagaacac cggaacaaag gccggaagga ctccaagcag ccgcacacct
     4861 gatttgtagc cagtgagttc gcacctaact ccaataaagt gcagaataaa tcctgcatta
     4921 acacactgag agtcgggcct tggacggccg attatagtcc ttttgtcgcc ataaaaagtc
     4981 ccctcgaggt cacgtgcgcg aaggaacccc ccctgagcag cttttcgccc agaaatcaag
     5041 gaaggggcgg caccagcagg caccggcggc cactagtccg tgcggcgggg tctgtgacat
     5101 tcaggggcca gcgtgccttt agtgatatcg ccgcagcgcg caaactcgga gaggaccact
     5161 ccgagcagaa gttagttaag caattctcta cgctaaaaaa aaaagtaaag tggattaagc
     5221 tcctactgga gagttaagta tgtgtacact tccagatatg cacaacgtgc taggcaacta
     5281 ttgttctaag gagagatgtg gtgcggagta aaatgcggag gtagtatggg gcttgcagac
     5341 atatataaag tctagtcatt cggctgaaaa taaactagaa actacttctt ccctttccat
     5401 ctacatccca ctcatccaac aatatcaaca atacaatagt aatataatat acaca
//
