LOCUS       sequence-172827-        8097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 under control of GAL1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8097)
  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 8097)
  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: pHES834
FEATURES             Location/Qualifiers
     source          1..8097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        314..501
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        2143..2584
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2143..2260
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2555..2578
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2602..3300
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     primer_bind     complement(3302..3318)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3302..3318)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4644..4663)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4645..4663)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4670..4687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4670..4686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4679..4701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4827..5282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4914..4933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5124..5145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5638..5657)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5757..5779
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5817..5835)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5920..6024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6025..6093,6094..6885)
                     /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(6243..6262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7056..7644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7545..7564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7798..7815
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7932..7953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7968..7973,7974..7991,7992..7998)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8006..8022
                     /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     8011..8033
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8030..8046
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8030..8046
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8067..8085
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgct gcacggtcct gttccctagc
       61 atgtacgtga gcgtatttcc ttttaaacca cgacgctttg tcttcattca acgtttccca
      121 ttgttttttt ctactattgc tttgctgtgg gaaaaactta tcgaaagatg acgacttttt
      181 cttaattctc gttttaagag cttggtgagc gctaggagtc actgccaggt atcgtttgaa
      241 cacggcatta gtcagggaag tcataacaca gtcctttccc gcaattttct ttttctatta
      301 ctcttggcct cctctagtac actctatatt tttttatgcc tcggtaatga ttttcatttt
      361 tttttttcca cctagcggat gactcttttt ttttcttagc gattggcatt atcacataat
      421 gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag
      481 gcaagataaa cgaaggcaaa gaggctctat atgtatataa agctgcggat gccttctgat
      541 ttaatctagt tctagtttac ttcaggttta cttaaagttg agataattga agcaaagaga
      601 agcatcgagc aaaggatggc gtttgttaag agggttacgc aggagacgaa tatacagctg
      661 gcgctggatc ttgacggtgg gtctgtttct gtacgggaga gcatactggg caaggaatat
      721 gctagtggtg atgggcagac catccatgtg cacactggag ttgggttttt ggaccacatg
      781 ttgactgcgc tggcgaagca tggcgggtgg tctctgatcc tggagtgtat aggggacttg
      841 cacatcgatg accaccacac tgttgaggac tgtgggattg cgctgggcca agcgttcaag
      901 gaggcgcttg gctccgtgcg tggtatcaag aggttcgggc atgggtttgc accactggac
      961 gaggcgctga gccgcgctgt ggttgacttc tccaataggc ctttcgccgt ggtggagctg
     1021 ggcctgaagc gagaacgcat aggccagcta tccacagaga tgatcccgca cttcttggag
     1081 agtttcgcca ctgaggcgcg tatcaccatg catgtggact gtctgcgggg caccaacgac
     1141 caccaccgct ccgaatcagc tttcaaggcg ctcgccatcg ccatcagaga ggcaagaaca
     1201 cctacgggtc gcgatgacgt tccatccact aagggtgtcc tagcatagaa cacagcccac
     1261 agctaccacc atcaacaata tttatataga taacgtacac atagaaatca cacaaacaga
     1321 gtatttattc ttaactacat gaactaccat cagaccgtct gggccactat ataatgtgct
     1381 attcataaac gtgatcactc tacgtagcag gcgaccccag gtgaaaattt ttcagcgagc
     1441 tgccagattg tcaggtgaaa aactgaaaaa aacttctggg cgatgagctt gtggcgggaa
     1501 attaagtata taaagcagtt agtttcttct gcttcttgtg gttctgggta ttcttgtaat
     1561 atttggaata atagaaaacc aagactactg cctttctttt catattatag aggaagaaat
     1621 acgcacgaac acgatataga ggtaaatttt ttgcaaaaag ataccaaact ataaaagaaa
     1681 tagaagacta aactacaaaa aataaagaaa gtaatacata tatccaatat gactgagtta
     1741 gcagagcaag tttccatgcc tgaagggcaa caaagacgtg gtggttacgt tccacctcac
     1801 atccgtaacg gtggtggcag aagaggcgat tctgctcctc gtggtggttt caacaacggt
     1861 ggcggccgtg gtggtttctt cagaaacaac ggtgaccgtg gtggattcgg ctttgggtac
     1921 cgggccctta tattgaattt tcaaaaattc ttactttttt tttggatgga cgcaaagaag
     1981 tttaataatc atattacatg gcattaccac catatacata tccatatcta atcttactta
     2041 tatgttgtgg aaatgtaaag agccccatta tcttagccta aaaaaacctt ctctttggaa
     2101 ctttcagtaa tacgcttaac tgctcattgc tatattgaag tacggattag aagccgccga
     2161 gcgggcgaca gccctccgac ggaagactct cctccgtgcg tcctcgtctt caccggtcgc
     2221 gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg aacaataaag attctacaat
     2281 actagctttt atggttatga agaggaaaaa ttggcagtaa cctggcccca caaaccttca
     2341 aattaacgaa tcaaattaac aaccatagga tgataatgcg attagttttt tagccttatt
     2401 tctggggtaa ttaatcagcg aagcgatgat ttttgatcta ttaacagata tataaatgga
     2461 aaagctgcat aaccacttta actaatactt tcaacatttt cagtttgtat tacttcttat
     2521 tcaaatgtca taaaagtatc aacaaaaaat tgttaatata cctctatact ttaacgtcaa
     2581 ggagaaaaaa ctatactcga gatggtgagc gagctgatta aggagaacat gcacatgaag
     2641 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
     2701 aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc
     2761 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac
     2821 acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga
     2881 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     2941 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     3001 atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc cgctgacggc
     3061 ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     3121 aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     3181 tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     3241 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagataa
     3301 ggatccacta gttctagaca cctgcaacat gcgtgagtaa ataagcggcc gccaccgcgg
     3361 tggagctcta agcaaatagc taaattatat acgaattaat attatgatta agtgtttacg
     3421 tgagtgcgat atttttatta ctatcttata cagttgtata tactctataa aatgagttgt
     3481 ctattaatta acgcgatgaa tgctttctgg gtttacctct ccaacaactc tagtttactt
     3541 ctcaatacat tcaattgtat ttgatttgtc aatacttcat cattaatcaa ttctatagtt
     3601 ttgtttttct cgtttatttc caaatttaat gcatcaattt tattattcaa tttgtcgttg
     3661 attttggtta atgattttat ggtttgatct ctggcattga ttgtttgtgt tagtttttca
     3721 ttattgataa ttaaattatt taagttagtt atcaactcgg tgttttcaag tttcaagttt
     3781 tcaatttctt tagagtttat tagatttgtc aaagtttctg aattgcttga ttggtcctgt
     3841 agaagagtat ttgttgttgt ggataattga ttcaattttt gagacaattg ctggaaggcg
     3901 ttgaaatatc tagcatcaat ctcatggttt ttttcccgag agtctcgtag attcaattgt
     3961 tttaatatat cttgggacca ctcttgattt gaactcatgg aaattaaact gggtgttgtg
     4021 ttgtggtgta atgattgtac cccctttgct tataattgtg tggtgacacc gattatttaa
     4081 agctgcagca tacgatatat atacatgtgt atatatgtat acctatgaat gtcagtaagt
     4141 atgtatacga acagtatgat actgaagatg acaaggtaat gcatcattct atacgtgtca
     4201 ttctgaacga ggcgcgcttt ccttttttct ttttgctttt tctttttttt tctcttgaac
     4261 tccagaaaaa aaatataaaa gagatggagg aacgggaaaa agttagttgt ggtgataggt
     4321 ggcaagtggt attccgtaag aacaacaaga aaagcatttc atattatggc tgaactgagc
     4381 gaacaagtgc aaaatttaag catcaacgac aacaacgaga atggttatgt tcctcctcac
     4441 ttaagaggaa aaccaagaag tgccagaaat aacagtagca actacaataa caacaacggc
     4501 ggctacaacg gtggccgtgg cggtggcagc ttctttagca acaaccgtcg tggtggttac
     4561 ggcaacggtg gtttcttcgg tggaaacaac ggtggcagcg gccggccatt taaatgttta
     4621 aacggccacg ctggcccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     4681 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4741 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4801 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4861 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4921 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4981 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5041 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5101 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5161 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5221 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5281 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtaa
     5341 taactgatat aattaaattg aagctctaat ttgtgagttt agtattagga tctctaccct
     5401 tggcgaaaag tcctctgcca acaatgatga tatctgatcc accacttaca acttcgtcga
     5461 cggttctgta ctgctgaccc aatatgcgta tatataccaa tctaagtctg tgctccttcc
     5521 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc
     5581 aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac
     5641 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5701 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5761 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5821 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcggc
     5881 gcgccgagga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5941 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6001 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6061 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6121 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6181 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6241 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6301 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6361 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6421 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6481 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6541 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6601 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6661 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6721 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6781 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6841 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6901 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6961 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7021 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7081 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7141 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7201 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7261 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7321 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7381 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7441 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7501 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7561 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7621 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7681 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7741 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7801 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7861 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7921 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7981 cttccggctc ctatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8041 tatgaccatg attacgccaa gctcggaatt aaccctcact aaagggaaca aaagctg
//
