LOCUS       sequence-181032-        8071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP under control of GAL1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8071)
  AUTHORS   Aranda-Diaz A, Mace K, Zuleta I, Harrigan P, El-Samad H
  TITLE     Robust Synthetic Circuits for Two-Dimensional Control of Gene 
            Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251.
  PUBMED    27930885
REFERENCE   2  (bases 1 to 8071)
  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"; journalName: "ACS Synth 
            Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251."
COMMENT     Sequence Label: pHES833
FEATURES             Location/Qualifiers
     source          1..8071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        306..493
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        2101..2542
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     2513..2536
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2557..3273
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3275..3291)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3275..3291)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4610..4629)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4611..4629)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4636..4653)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4636..4652)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4645..4667)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4793..5248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4880..4899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5090..5111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5604..5623)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5723..5745
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5783..5801)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5886..5990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5991..6059,6060..6851)
                     /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     complement(6209..6228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7022..7610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7511..7530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7764..7781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7898..7919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7934..7939,7940..7957,7958..7964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7972..7988
                     /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)."
     primer_bind     7977..7999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7996..8012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7996..8012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8033..8051
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atttaaatgt ttaaacggcc acgctggccg ctgcacggtc ctgttcccta gcatgtacgt
       61 gagcgtattt ccttttaaac cacgacgctt tgtcttcatt caacgtttcc cattgttttt
      121 ttctactatt gctttgctgt gggaaaaact tatcgaaaga tgacgacttt ttcttaattc
      181 tcgttttaag agcttggtga gcgctaggag tcactgccag gtatcgtttg aacacggcat
      241 tagtcaggga agtcataaca cagtcctttc ccgcaatttt ctttttctat tactcttggc
      301 ctcctctagt acactctata tttttttatg cctcggtaat gattttcatt tttttttttc
      361 cacctagcgg atgactcttt ttttttctta gcgattggca ttatcacata atgaattata
      421 cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc aggcaagata
      481 aacgaaggca aagaggctct atatgtatat aaagctgcgg atgccttctg atttaatcta
      541 gttctagttt acttcaggtt tacttaaagt tgagataatt gaagcaaaga gaagcatcga
      601 gcaaaggatg gcgtttgtta agagggttac gcaggagacg aatatacagc tggcgctgga
      661 tcttgacggt gggtctgttt ctgtacggga gagcatactg ggcaaggaat atgctagtgg
      721 tgatgggcag accatccatg tgcacactgg agttgggttt ttggaccaca tgttgactgc
      781 gctggcgaag catggcgggt ggtctctgat cctggagtgt ataggggact tgcacatcga
      841 tgaccaccac actgttgagg actgtgggat tgcgctgggc caagcgttca aggaggcgct
      901 tggctccgtg cgtggtatca agaggttcgg gcatgggttt gcaccactgg acgaggcgct
      961 gagccgcgct gtggttgact tctccaatag gcctttcgcc gtggtggagc tgggcctgaa
     1021 gcgagaacgc ataggccagc tatccacaga gatgatcccg cacttcttgg agagtttcgc
     1081 cactgaggcg cgtatcacca tgcatgtgga ctgtctgcgg ggcaccaacg accaccaccg
     1141 ctccgaatca gctttcaagg cgctcgccat cgccatcaga gaggcaagaa cacctacggg
     1201 tcgcgatgac gttccatcca ctaagggtgt cctagcatag aacacagccc acagctacca
     1261 ccatcaacaa tatttatata gataacgtac acatagaaat cacacaaaca gagtatttat
     1321 tcttaactac atgaactacc atcagaccgt ctgggccact atataatgtg ctattcataa
     1381 acgtgatcac tctacgtagc aggcgacccc aggtgaaaat ttttcagcga gctgccagat
     1441 tgtcaggtga aaaactgaaa aaaacttctg ggcgatgagc ttgtggcggg aaattaagta
     1501 tataaagcag ttagtttctt ctgcttcttg tggttctggg tattcttgta atatttggaa
     1561 taatagaaaa ccaagactac tgcctttctt ttcatattat agaggaagaa atacgcacga
     1621 acacgatata gaggtaaatt ttttgcaaaa agataccaaa ctataaaaga aatagaagac
     1681 taaactacaa aaaataaaga aagtaataca tatatccaat atgactgagt tagcagagca
     1741 agtttccatg cctgaagggc aacaaagacg tggtggttac gttccacctc acatccgtaa
     1801 cggtggtggc agaagaggcg attctgctcc tcgtggtggt ttcaacaacg gtggcggccg
     1861 tggtggtttc ttcagaaaca acggtgaccg tggtggattc ggctttgggt accgggccct
     1921 tggatggacg caaagaagtt taataatcat attacatggc attaccacca tatacatatc
     1981 catatctaat cttacttata tgttgtggaa atgtaaagag ccccattatc ttagcctaaa
     2041 aaaaccttct ctttggaact ttcagtaata cgcttaactg ctcattgcta tattgaagta
     2101 cggactagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
     2161 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcacgccaca ctgctccgaa
     2221 caataaagat tctacaatac tagcttttat agttatgaag aggaaaaatt ggcagtaacc
     2281 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
     2341 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
     2401 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
     2461 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
     2521 tctatacttt aacgtcaagg agaactcgag aataaaatgt ctaaaggtga agaattattc
     2581 actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct
     2641 gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa attgatttgt
     2701 actactggta aattgccagt tccatggcca accttagtca ctactttagg ttatggtttg
     2761 caatgttttg ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg
     2821 ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc
     2881 agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt
     2941 gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac
     3001 aatgtttaca tcactgctga caaacaaaag aatggtatca aagctaactt caaaattaga
     3061 cacaacattg aagatggtgg tgttcaatta gctgaccatt atcaacaaaa tactccaatt
     3121 ggtgatggtc cagtcttgtt accagacaac cattacttat cctatcaatc tgccttatcc
     3181 aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt
     3241 attacccatg gtatggatga attgtacaaa taaggatcca ctagttctag acacctgcaa
     3301 catgcgtgag taaataagcg gccgccaccg cggtggagct ctaagcaaat agctaaatta
     3361 tatacgaatt aatattatga ttaagtgttt acgtgagtgc gatattttta ttactatctt
     3421 atacagttgt atatactcta taaaatgagt tgtctattaa ttaacgcgat gaatgctttc
     3481 tgggtttacc tctccaacaa ctctagttta cttctcaata cattcaattg tatttgattt
     3541 gtcaatactt catcattaat caattctata gttttgtttt tctcgtttat ttccaaattt
     3601 aatgcatcaa ttttattatt caatttgtcg ttgattttgg ttaatgattt tatggtttga
     3661 tctctggcat tgattgtttg tgttagtttt tcattattga taattaaatt atttaagtta
     3721 gttatcaact cggtgttttc aagtttcaag ttttcaattt ctttagagtt tattagattt
     3781 gtcaaagttt ctgaattgct tgattggtcc tgtagaagag tatttgttgt tgtggataat
     3841 tgattcaatt tttgagacaa ttgctggaag gcgttgaaat atctagcatc aatctcatgg
     3901 tttttttccc gagagtctcg tagattcaat tgttttaata tatcttggga ccactcttga
     3961 tttgaactca tggaaattaa actgggtgtt gtgttgtggt gtaatgattg tacccccttt
     4021 gcttataatt gtgtggtgac accgattatt taaagctgca gcatacgata tatatacatg
     4081 tgtatatatg tatacctatg aatgtcagta agtatgtata cgaacagtat gatactgaag
     4141 atgacaaggt aatgcatcat tctatacgtg tcattctgaa cgaggcgcgc tttccttttt
     4201 tctttttgct ttttcttttt ttttctcttg aactccagaa aaaaaatata aaagagatgg
     4261 aggaacggga aaaagttagt tgtggtgata ggtggcaagt ggtattccgt aagaacaaca
     4321 agaaaagcat ttcatattat ggctgaactg agcgaacaag tgcaaaattt aagcatcaac
     4381 gacaacaacg agaatggtta tgttcctcct cacttaagag gaaaaccaag aagtgccaga
     4441 aataacagta gcaactacaa taacaacaac ggcggctaca acggtggccg tggcggtggc
     4501 agcttcttta gcaacaaccg tcgtggtggt tacggcaacg gtggtttctt cggtggaaac
     4561 aacggtggca gcggccggcc atttaaatgt ttaaacggcc acgctggccc cctatagtga
     4621 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     4681 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     4741 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     4801 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4861 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4921 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4981 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5041 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5101 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5161 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5221 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     5281 gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt
     5341 gagtttagta ttaggatctc tacccttggc gaaaagtcct ctgccaacaa tgatgatatc
     5401 tgatccacca cttacaactt cgtcgacggt tctgtactgc tgacccaata tgcgtatata
     5461 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc
     5521 aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa
     5581 aaggtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5641 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5701 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5761 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5821 ataataatgg tttcttagac gtcggcgcgc cgaggacgtc aggtggcact tttcggggaa
     5881 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5941 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6001 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6061 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6121 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6181 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6241 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6301 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6361 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6421 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6481 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6541 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6601 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6661 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6721 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6781 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6841 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6901 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7141 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7801 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7861 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7921 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7981 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc ggaattaacc
     8041 ctcactaaag ggaacaaaag ctgggccggc c
//
