LOCUS       sequence-180370-        7906 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 7906)
  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 7906)
  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: pHES820
FEATURES             Location/Qualifiers
     source          1..7906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    110..131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(146..151,152..169,170..176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    184..200
                     /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     189..211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     208..224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     208..224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        245..263
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2403..2844
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     2815..2838
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2859..3575
                     /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(3577..3593)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3577..3593)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4728..4747)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4729..4747)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4754..4771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4754..4770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4763..4785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4911..5366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4998..5017)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5208..5229
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5722..5741)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5841..5863
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5901..5919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6004..6108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6109..6177,6178..6969)
                     /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(6327..6346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7140..7728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7629..7648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7882..7899
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
       61 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      121 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      181 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      241 tcggaattaa ccctcactaa agggaacaaa agctgggccg gccatttaaa tgtttaaacg
      301 gccacgctgg ccatatttct accaatctct caacactgag taatggtagt tataagaaag
      361 agaccgagtt agggacagtt agaggcggtg gagatattcc ttatggcatg tctggcgatg
      421 ataaaacttt tcaaacggca gccccgatct aaaagagctg acagggaaat ggtcagaaaa
      481 agaaacgtgc acccgcccgt ctggacgcgc cgctcacccg cacggcagag accaatcagt
      541 aaaaatcaac ggttaacgac attactatat atataatata ggaagcattt aatagaacag
      601 catcgtaata tatgtgtact ttgcagttat gacgccagat ggcagtagtg gaagatattc
      661 tttattgaaa aatagcttgt caccttacgt acaatcttga tccggagccc gtcgctttga
      721 gagaaatcag aagattccaa aagtctactg aacttttgat cagaaagttg cctttccaaa
      781 gactagtcag agaaatcgcc caagatttca agaccgatct aagattccaa tcctctgcca
      841 tcggtgctct acaggaatcc gttgaagctt acttggtctc tttgttcgaa gacaccaact
      901 tggccgctat ccacgctaag cgtgttacca tccaaaagaa ggatatcaag ttggctagaa
      961 gattgagagg tgaaagatcc taagcacaca gggattgttg tacaattact ttttatgatt
     1021 ttagatacac acaaatcttt tctacgatta cctttcattt attttcgttt catgttttat
     1081 taatcattct ttttgtatat ttcacatcgg gtggggtttt tatattaaat tttaataaca
     1141 taatctttta aagtttctac tttctatcct gtatattatt tccatatcta tatctatatt
     1201 ttcaagcttg ttcacatcct tatcggaatt gttccagcgt ttttgtacag aaaatgatgt
     1261 tatggtaacg agctatatca gacctactag tcttatagag ctgtacagag tcttggaaac
     1321 ataaccaaca caccaacaag atgtcatttg attcgttact cgacaagaat gataagctgg
     1381 taaaagtttg cgggattcaa accgtcgagg ctgccgaaac tgcgcttcaa gcaggcgctg
     1441 atttgatagg gatcatatgt gtccccaaca ggaagcggac tatcgagagc gctgtggctc
     1501 gtgaaatatc caaattgatt cacaaatcag gtactacaaa gctggtgggg gtgttcagga
     1561 atcaatctgt tgaggacgta catcggcttt ctgaggaata tgaccttgac ataatccaat
     1621 tacatggtga tgaatcatgg ccagagtact ataacgtcat taagaaacca ataatcaaaa
     1681 gagtcatatt ccctagagat gtcgatgttg taacacaagt gtgtcaaaga aaacccttgg
     1741 tatgtctacc attgttcgac tccgaggcag gcggtacagg tgaaaagctg gactggtcca
     1801 gtatttcctc ctgggcaagc gaacagaata acgtatttta tatactagcg ggtggactaa
     1861 ctgcagaaaa cgtcatggag gcggtggggt tgcctggtgt aattggtgtt gacgtctcag
     1921 gtggtgtaga gactgacggc gtcaaagaca atgacaagat aattaaatat gtacaaaatg
     1981 caaagaaaca atgaattacg aagagaagtc tcaatgaggt tttacaaaca atcctataat
     2041 acatatatgc cacttagata tactttgaat actgaatata ctgactcatt atattcttta
     2101 attgtagttt ctcaaaaata ttcatacagg ttagctatga attcatgcta taatttccat
     2161 tttgttcatg taagatacac atagggataa aatgtgataa ctaatcagcg gtaccgggcc
     2221 cttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac catatacata
     2281 tccatatcta atcttactta tatgttgtgg aaatgtaaag agccccatta tcttagccta
     2341 aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc tatattgaag
     2401 tacggactag aagccgccga gcgggcgaca gccctccgac ggaagactct cctccgtgcg
     2461 tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcacgcca cactgctccg
     2521 aacaataaag attctacaat actagctttt atagttatga agaggaaaaa ttggcagtaa
     2581 cctggcccca caaaccttca aattaacgaa tcaaattaac aaccatagga tgataatgcg
     2641 attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat ttttgatcta
     2701 ttaacagata tataaatgga aaagctgcat aaccacttta actaatactt tcaacatttt
     2761 cagtttgtat tacttcttat tcaaatgtca taaaagtatc aacaaaaaat tgttaatata
     2821 cctctatact ttaacgtcaa ggagaactcg agaataaaat gtctaaaggt gaagaattat
     2881 tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt
     2941 ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaattgattt
     3001 gtactactgg taaattgcca gttccatggc caaccttagt cactacttta ggttatggtt
     3061 tgcaatgttt tgctagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca
     3121 tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga
     3181 ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta
     3241 ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc
     3301 acaatgttta catcactgct gacaaacaaa agaatggtat caaagctaac ttcaaaatta
     3361 gacacaacat tgaagatggt ggtgttcaat tagctgacca ttatcaacaa aatactccaa
     3421 ttggtgatgg tccagtcttg ttaccagaca accattactt atcctatcaa tctgccttat
     3481 ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg
     3541 gtattaccca tggtatggat gaattgtaca aataaggatc cactagttct agacacctgc
     3601 aacatgcgtg agtaaataag cggccgccac cgcggtggag ctctaagcaa atagctaaat
     3661 tatatacgaa ttaatattat gattaagtgt ttacgtgagt gcgatatttt tattactatc
     3721 ttatacagtt gtatatactc tataaaatga gttgtctatt aattaacgcg atgaatgctt
     3781 tctgggttta cctctccaac aactctagtt tacttctcaa tacattcaat tgtatttgat
     3841 ttgtcaatac ttcatcatta atcaattcta tagttttgtt tttctcgttt atttccaaat
     3901 ttaatgcatc aattttatta ttcaatttgt cgttgatttt ggttaatgat tttatggttt
     3961 gatctctggc attgattgtt tgtgttagtt tttcattatt gataattaaa ttatttaagt
     4021 tagttatcaa ctcggtgttt tcaagtttca agttttcaat ttctttagag tttattagat
     4081 ttgtcaaagt ttctgaattg cttgattggt cctgtagaag agtatttgtt gttgtggata
     4141 attgattcaa tttttgagac aattgctgga aggcgttgaa atatctagca tcaatctcat
     4201 ggtttttttc ccgagagtct cgtagattca attgttttaa tatatcttgg gaccactctt
     4261 gatttgaact catggaaatt aaactgggtg ttgtgttgtg gtgtaatgat tgtaccccct
     4321 ttgcttataa ttgtgtggtt ccgatgctga cttactgggt attatatgtg tgcccaatag
     4381 aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa
     4441 aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga
     4501 tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc
     4561 gtggcgagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa
     4621 agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt
     4681 tgattcagaa ggccggccat ttaaatgttt aaacggccac gctggccccc tatagtgagt
     4741 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4801 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4861 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     4921 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4981 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5041 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5101 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5161 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5221 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5281 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5341 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     5401 ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc taatttgtga
     5461 gtttagtatt aggatctcta cccttggcga aaagtcctct gccaacaatg atgatatctg
     5521 atccaccact tacaacttcg tcgacggttc tgtactgctg acccaatatg cgtatatata
     5581 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     5641 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     5701 ggtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5761 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5821 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5881 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5941 aataatggtt tcttagacgt cggcgcgccg aggacgtcag gtggcacttt tcggggaaat
     6001 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6061 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6121 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6181 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6241 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6301 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6361 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6421 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6481 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6541 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6601 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6661 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6721 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6781 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6841 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6901 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6961 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7021 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7261 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7741 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7801 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7861 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcgg
//
