LOCUS       sequence-201478-        5445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces uvarum 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: pILGFP4R
FEATURES             Location/Qualifiers
     source          1..5445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..732
                     /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(977..993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(977..993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(990..1012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1001..1017
                     /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(1025..1031,1032..1049,1050..1055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1070..1091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1208..1225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1379..1967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1459..1478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2138..2929,2930..2998))
                     /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     2761..2780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2999..3103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3171..3189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3227..3249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3349..3368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3562..3584
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3576..3593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3577..3593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4707..4740)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 aatagataaa aaatgggatc ctctaaaggt gaagaattat tcactggtgt tgtcccaatt
       61 ttggttgaat tagatggtga tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa
      121 ggtgatgcta cttacggtaa attgacctta aaatttattt gtactactgg taaattgcca
      181 gttccatggc caaccttagt cactactttc ggttatggtg ttcaatgttt tgcgagatac
      241 ccagatcata tgaaacaaca tgactttttc aagtctgcca tgccagaagg ttatgttcaa
      301 gaaagaacta tttttttcaa agatgacggt aactacaaga ccagagctga agtcaagttt
      361 gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa agaagatggt
      421 aacattttag gtcacaaatt ggaatacaac tataactctc acaatgttta catcatggct
      481 gacaaacaaa agaatggtat caaagttaac ttcaaaatta gacacaacat tgaagatggt
      541 tctgttcaat tagctgacca ttatcaacaa aatactccaa ttggtgatgg tccagtcttg
      601 ttaccagaca accattactt atccactcaa tctgccttat ccaaagatcc aaacgaaaag
      661 agagaccaca tggtcttgtt agaatttgtt actgctgctg gtattaccca tggtatggat
      721 gaattgtaca aaagatctta aactagtagg actatttgca aagggaaggg atgctaaggt
      781 agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca
      841 aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc
      901 aatttaatta tatcagttat tacccgggaa tctcggtcgt aatgatttaa atgcatgcaa
      961 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1021 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1081 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1201 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1261 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1321 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1381 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1441 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1501 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1561 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1621 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1681 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1741 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1801 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1861 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1921 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1981 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2041 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2101 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2221 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2281 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3121 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3181 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3241 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3301 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     3361 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3421 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     3481 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     3541 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattca
     3601 tttaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     3661 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     3721 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaaaca tgtccacaaa
     3781 atcatatacc agtagagctg agactcatgc aagtccggtt gcatcgaaac ttttacgttt
     3841 aatggatgaa aagaagacca atttgtgtgc ttctcttgac gttcgttcga ctgatgagct
     3901 attgaaactt gttgaaacgt tgggtccata catttgcctt ttgaaaacac acgttgatat
     3961 cttggatgat ttcagttatg agggtactgt cgttccattg aaagcattgg cagagaaata
     4021 caagttcttg atatttgagg acagaaaatt cgccgatatc ggtaacacag tcaaattaca
     4081 atatacatcg ggcgtttacc gtatcgcaga atggtctgat atcaccaacg cccacggggt
     4141 tactggtgct ggtattgttg ctggcttgaa acaaggtgcg caagaggtca ccaaagaacc
     4201 aaggggatta ttgatgcttg ctgaattgtc ttccaagggt tctctagcac acggtgaata
     4261 tactaagggt accgttgata ttgcaaagag tgataaagat ttcgttattg ggttcattgc
     4321 tcagaacgat atgggaggaa gagaagaagg gtttgattgg ctaatcatga ccccaggtgt
     4381 aggtttagac gacaaaggcg atgcattggg tcagcagtac agaaccgtcg acgaagttgt
     4441 aagtggtgga tcagatatca tcattgttgg cagaggactt ttcgccaagg gtagagatcc
     4501 taaggttgaa ggtgaaagat acagaaatgc tggatgggaa gcgtaccaaa agagaatcag
     4561 cgctccccat taattataca ggaaacttaa tagaacaaat cacatattta atctaatagc
     4621 cacctgcatt ggcacggtgc aacactactt caacttcatc ttacaaaaag atcacgtgat
     4681 ctgttgtatt gggatctcta gacctaataa cttcgtatag catacattat acgaagttat
     4741 attaagggtt gctcgagaaa agaaggggct ttccaaaaac cttaatttgt tggcgcaacc
     4801 aaagtgtttt ccagctggct aatggttcat atgctcaagt gtcctcaaaa cgtttacaag
     4861 ctaaaaagga caagcccccc acgcaagcgt gggaactgcc gactgaggcc aatgccctgt
     4921 attagttcat gcaaagatat caacaccatc ctttttctta cctttgtaaa ttaatctcac
     4981 atacagaaga agtcatgctc ctacgggtgt tttgtggccg cagcccttgc cacgcgggcg
     5041 aagatcttcc gagcggacgt ctctcgaccg aagtggcccc ttagtgggca aacgtcctag
     5101 atgttccggt acgccgatac tgttcctgaa aaatgtggag tttctccgga aaagagacat
     5161 gttctttcgg gagacagtgc tgcggtgaca tgaaatagta tcacataagc tcaggatgcc
     5221 tatgcaagtg gggattttta ttgaagcatt gtatgtggga gggatgtgtc tggcgtaacc
     5281 ttgttttgat ataaaacgaa cggctgcatg ctgcagtctt gagcttttct tgcttattca
     5341 attcttgttc tacttggtaa atagccgtca agaaaaatac aaggtaatct ctccccttag
     5401 ttattgcatt attctaaata aatcaaagag aagatttttt tgaag
//
