LOCUS       sequence-180719-        8641 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 8641)
  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 8641)
  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: pHES840
FEATURES             Location/Qualifiers
     source          1..8641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..56)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        141..245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(246..314,315..1106)
                     /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(464..483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1277..1865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1766..1785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2019..2036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2153..2174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2189..2194,2195..2212,2213..2219)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2227..2243
                     /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     2232..2254
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2251..2267
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2251..2267
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2288..2306
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2594..3001
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        5008..5449
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     5420..5443
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5464..6180
                     /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(6182..6198)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6182..6198)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7506..7525)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7507..7525)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7532..7549)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7532..7548)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7541..7563)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7689..8144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7776..7795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7986..8007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8500..8519)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8619..8641
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
       61 taggttaatg tcatgataat aatggtttct tagacgtcgg cgcgccgagg acgtcaggtg
      121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      481 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      781 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      901 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2041 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2101 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2161 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2221 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2281 agctcggaat taaccctcac taaagggaac aaaagctggg ccggccattt aaatgtttaa
     2341 acggccacgc tggccgagga gaacttctag tatatctaca tacctaatat tattgcctta
     2401 ttaaaaatgg aatcccaacg attacatcaa aatccacatt ctcttcaaaa tcaattgtcc
     2461 tgtacttcct tgttcatgtg tgttcaaaaa cgttatattt ataggataat tatactctat
     2521 ttctcaacaa gtaattggtt gtttggccga gcggtctaag gcgcctgatt caagaaatat
     2581 cttgaccgca gttaactgtg ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc
     2641 ttagcaacca ttattttttt cctcaacata acgagaacac acaggggcgc tatcgcacag
     2701 aatcaaattc gatgactgga aattttttgt taatttcaga ggtcgcctga cgcatatacc
     2761 tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac cggcttttca
     2821 tatagaatag agaagcgttc atgactaaat gcttgcatca caatacttga agttgacaat
     2881 attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt actttctaac
     2941 ttttcttacc ttttacattt cagcaatata tatatatata tttcaaggat ataccattct
     3001 aatgtctgcc cctaagttac caatagatag cggtttaata taaggtttca acaagaaata
     3061 ttttgtgaaa atcaaaagtt aatagtttca ctgtcgaaaa tatgaaagtt taagttttca
     3121 gttcatccca ttgtagttat caaatttctg tgtttcccgt atatgtgtgc tctgtattca
     3181 tcgctatctt gaaactggtt atgatcgttg cgcatgtgac ctgtatcgct caaaagagcc
     3241 cggcaccggt accgggcaaa agcccctagc aagctaaaat gattcaactt tcgtagtaat
     3301 ttgttcaacc ggtcacagta aagtcaatga ctaacttttg aatcgaggat gtatttccaa
     3361 ttgaaagcaa tgtatatata aaccattgtg ttgaagcttg tcttacagtt ttcttgtatc
     3421 ctcccattgt tcatatagta cttaaagtac cataatcctt aaagaacaaa cttgaaagag
     3481 aatacactat acacaactaa taaataaaga ataatcatgg ctgtgaccaa gacaattgta
     3541 gttctaccag gtgaccatgt tggtcaagaa atcactgaag aggccattaa agttttaaat
     3601 gctattcagg aatgtcgtcc agacaaggtc aatttcaagt ttgatcatca tttgatcggt
     3661 ggtgctgcaa ttgatgccac tggtgttcca ttaccagacg aagctttgga ggcctctaag
     3721 aaagctgatg ccgtgcttct gggtgctgtt ggtggtccaa aatggggtac tggcgctgtc
     3781 agaccagaac aaggtctttt gaaaatccgt aaggagttgc aattgtatgc taatctaaga
     3841 ccatgtaatt ttgcatctga ttccttacta gatctatcgc cattgaagcc tgaaattgca
     3901 agaggtacag atttcgtcgt tgttagagaa ctagtgggtg gtatttattt cggggagaga
     3961 aaagaagatg aaggtgatgg tgtcgcctgg gatagcgaaa agtattctgt gcccgaagtt
     4021 caaagaatca ctagaatggc tgctttcatg gctctgcaac acaacccacc tttacctatt
     4081 tggtctctag ataaagccaa cgttctagca tcatcaagat tatggagaaa aacagtggaa
     4141 gagacgatta agaatgaatt cccacaattg actgttcaac atcaattgat tgattccgct
     4201 gctatgatct tggttaagaa cccaactcat ctaaatggta ttattatcac aaacaatatg
     4261 tttggtgata ttatctccga tgaagcctcc gtgatcccag gttcgttagg attacttcca
     4321 tctgcctcct tggcatctct tccagataag aatacggcat ttggtttata tgagccatgt
     4381 catggctctg ctccagactt accaaaggga aaggttaacc ccgtagcaac aatcctatct
     4441 gctgctatga tgttgaagtt gtctctggac ctgttcgaag aaggtgaaat tatagaacaa
     4501 gcagtcaaga aagtgttgga ttctggtatc agaacagcgg atctaaaggg taccaactct
     4561 actactgagg tcggtgatgc agtagcgaaa gctgtcaggg aactattagc ttagacttag
     4621 tagatgtaca gttgcataca taagtctgaa cagaaagaag taaatatcat atctagttcg
     4681 gtttagaaat cagtgactat attggttttg accagatcgc aatccttgca agcatttagt
     4741 aatgactatc aaaccggtga atcatctgat aaggctatat tagcttgtgc attcgcatgt
     4801 atcgggaaac gaacggtacc gggcccttgg atggacgcaa agaagtttaa taatcatatt
     4861 acatggcatt accaccatat acatatccat atctaatctt acttatatgt tgtggaaatg
     4921 taaagagccc cattatctta gcctaaaaaa accttctctt tggaactttc agtaatacgc
     4981 ttaactgctc attgctatat tgaagtacgg actagaagcc gccgagcggg cgacagccct
     5041 ccgacggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag
     5101 atgtgcctca cgccacactg ctccgaacaa taaagattct acaatactag cttttatagt
     5161 tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatta acgaatcaaa
     5221 ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg ggtaattaat
     5281 cagcgaagcg atgatttttg atctattaac agatatataa atggaaaagc tgcataacca
     5341 ctttaactaa tactttcaac attttcagtt tgtattactt cttattcaaa tgtcataaaa
     5401 gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga actcgagaat
     5461 aaaatgtcta aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat
     5521 ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac
     5581 ggtaaattga ccttaaaatt gatttgtact actggtaaat tgccagttcc atggccaacc
     5641 ttagtcacta ctttaggtta tggtttgcaa tgttttgcta gatacccaga tcatatgaaa
     5701 caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt
     5761 ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta
     5821 gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac
     5881 aaattggaat acaactataa ctctcacaat gtttacatca ctgctgacaa acaaaagaat
     5941 ggtatcaaag ctaacttcaa aattagacac aacattgaag atggtggtgt tcaattagct
     6001 gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat
     6061 tacttatcct atcaatctgc cttatccaaa gatccaaacg aaaagagaga ccacatggtc
     6121 ttgttagaat ttgttactgc tgctggtatt acccatggta tggatgaatt gtacaaataa
     6181 ggatccacta gttctagaca cctgcaacat gcgtgagtaa ataagcggcc gccaccgcgg
     6241 tggagctcta agcaaatagc taaattatat acgaattaat attatgatta agtgtttacg
     6301 tgagtgcgat atttttatta ctatcttata cagttgtata tactctataa aatgagttgt
     6361 ctattaatta acgcgatgaa tgctttctgg gtttacctct ccaacaactc tagtttactt
     6421 ctcaatacat tcaattgtat ttgatttgtc aatacttcat cattaatcaa ttctatagtt
     6481 ttgtttttct cgtttatttc caaatttaat gcatcaattt tattattcaa tttgtcgttg
     6541 attttggtta atgattttat ggtttgatct ctggcattga ttgtttgtgt tagtttttca
     6601 ttattgataa ttaaattatt taagttagtt atcaactcgg tgttttcaag tttcaagttt
     6661 tcaatttctt tagagtttat tagatttgtc aaagtttctg aattgcttga ttggtcctgt
     6721 agaagagtat ttgttgttgt ggataattga ttcaattttt gagacaattg ctggaaggcg
     6781 ttgaaatatc tagcatcaat ctcatggttt ttttcccgag agtctcgtag attcaattgt
     6841 tttaatatat cttgggacca ctcttgattt gaactcatgg aaattaaact gggtgttgtg
     6901 ttgtggtgta atgattgtac cccctttgct tataattgtg tggccaccga agtcggtgat
     6961 gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat
     7021 ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga
     7081 atatgttcat agggtagacg aaactatata cgcaatctac atacatttat caagaaggag
     7141 aaaaaggagg atgtaaagga atacaggtaa gcaaattgat actaatggct caacgtgata
     7201 aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcacc acacaaaaag
     7261 ttaggtgtaa cagaaaatca tgaaactatg attcctaatt tatatattgg aggattttct
     7321 ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg acctgaccat ttgatggagt
     7381 ttaagtcaat acctccttga accatttccc ataatggtga aagttccctc aagaatttta
     7441 ctctgtcaga aacggcctta acgacgtagg ccggccattt aaatgtttaa acggccacgc
     7501 tggcccccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     7561 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     7621 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     7681 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     7741 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     7801 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     7861 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     7921 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     7981 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     8041 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     8101 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     8161 ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat
     8221 tgaagctcta atttgtgagt ttagtattag gatctctacc cttggcgaaa agtcctctgc
     8281 caacaatgat gatatctgat ccaccactta caacttcgtc gacggttctg tactgctgac
     8341 ccaatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt
     8401 cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa
     8461 aaaatgatga attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc
     8521 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     8581 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     8641 g
//
