LOCUS       sequence-180369-        8053 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 8053)
  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 8053)
  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..8053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        318..505
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        2113..2554
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     2525..2548
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2569..3267
                     /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(3269..3285)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3269..3285)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4604..4623)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4605..4623)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4630..4647)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4630..4646)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4639..4661)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4787..5242
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4874..4893)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5084..5105
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5598..5617)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5717..5739
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5777..5795)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5880..5984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5985..6053,6054..6845)
                     /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(6203..6222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7016..7604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7505..7524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7758..7775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7892..7913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7928..7933,7934..7951,7952..7958)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7966..7982
                     /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     7971..7993
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7990..8006
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7990..8006
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8027..8045
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gctgggccgg ccatttaaat gtttaaacgg ccacgctggc cgctgcacgg tcctgttccc
       61 tagcatgtac gtgagcgtat ttccttttaa accacgacgc tttgtcttca ttcaacgttt
      121 cccattgttt ttttctacta ttgctttgct gtgggaaaaa cttatcgaaa gatgacgact
      181 ttttcttaat tctcgtttta agagcttggt gagcgctagg agtcactgcc aggtatcgtt
      241 tgaacacggc attagtcagg gaagtcataa cacagtcctt tcccgcaatt ttctttttct
      301 attactcttg gcctcctcta gtacactcta tattttttta tgcctcggta atgattttca
      361 tttttttttt tccacctagc ggatgactct ttttttttct tagcgattgg cattatcaca
      421 taatgaatta tacattatat aaagtaatgt gatttcttcg aagaatatac taaaaaatga
      481 gcaggcaaga taaacgaagg caaagaggct ctatatgtat ataaagctgc ggatgccttc
      541 tgatttaatc tagttctagt ttacttcagg tttacttaaa gttgagataa ttgaagcaaa
      601 gagaagcatc gagcaaagga tggcgtttgt taagagggtt acgcaggaga cgaatataca
      661 gctggcgctg gatcttgacg gtgggtctgt ttctgtacgg gagagcatac tgggcaagga
      721 atatgctagt ggtgatgggc agaccatcca tgtgcacact ggagttgggt ttttggacca
      781 catgttgact gcgctggcga agcatggcgg gtggtctctg atcctggagt gtatagggga
      841 cttgcacatc gatgaccacc acactgttga ggactgtggg attgcgctgg gccaagcgtt
      901 caaggaggcg cttggctccg tgcgtggtat caagaggttc gggcatgggt ttgcaccact
      961 ggacgaggcg ctgagccgcg ctgtggttga cttctccaat aggcctttcg ccgtggtgga
     1021 gctgggcctg aagcgagaac gcataggcca gctatccaca gagatgatcc cgcacttctt
     1081 ggagagtttc gccactgagg cgcgtatcac catgcatgtg gactgtctgc ggggcaccaa
     1141 cgaccaccac cgctccgaat cagctttcaa ggcgctcgcc atcgccatca gagaggcaag
     1201 aacacctacg ggtcgcgatg acgttccatc cactaagggt gtcctagcat agaacacagc
     1261 ccacagctac caccatcaac aatatttata tagataacgt acacatagaa atcacacaaa
     1321 cagagtattt attcttaact acatgaacta ccatcagacc gtctgggcca ctatataatg
     1381 tgctattcat aaacgtgatc actctacgta gcaggcgacc ccaggtgaaa atttttcagc
     1441 gagctgccag attgtcaggt gaaaaactga aaaaaacttc tgggcgatga gcttgtggcg
     1501 ggaaattaag tatataaagc agttagtttc ttctgcttct tgtggttctg ggtattcttg
     1561 taatatttgg aataatagaa aaccaagact actgcctttc ttttcatatt atagaggaag
     1621 aaatacgcac gaacacgata tagaggtaaa ttttttgcaa aaagatacca aactataaaa
     1681 gaaatagaag actaaactac aaaaaataaa gaaagtaata catatatcca atatgactga
     1741 gttagcagag caagtttcca tgcctgaagg gcaacaaaga cgtggtggtt acgttccacc
     1801 tcacatccgt aacggtggtg gcagaagagg cgattctgct cctcgtggtg gtttcaacaa
     1861 cggtggcggc cgtggtggtt tcttcagaaa caacggtgac cgtggtggat tcggctttgg
     1921 gtaccgggcc cttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac
     1981 catatacata tccatatcta atcttactta tatgttgtgg aaatgtaaag agccccatta
     2041 tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc
     2101 tatattgaag tacggactag aagccgccga gcgggcgaca gccctccgac ggaagactct
     2161 cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcacgcca
     2221 cactgctccg aacaataaag attctacaat actagctttt atagttatga agaggaaaaa
     2281 ttggcagtaa cctggcccca caaaccttca aattaacgaa tcaaattaac aaccatagga
     2341 tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat
     2401 ttttgatcta ttaacagata tataaatgga aaagctgcat aaccacttta actaatactt
     2461 tcaacatttt cagtttgtat tacttcttat tcaaatgtca taaaagtatc aacaaaaaat
     2521 tgttaatata cctctatact ttaacgtcaa ggagaactcg agaataaaat ggtgagcgag
     2581 ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac
     2641 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc
     2701 aaggcggtcg agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg
     2761 tacggcagca aaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc
     2821 ttccccgagg gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc
     2881 gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg
     2941 gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggcctcc
     3001 accgagaccc tgtaccccgc tgacggcggc ctggaaggca gagccgacat ggccctgaag
     3061 ctcgtgggcg ggggccacct gatctgcaac ttgaagacca catacagatc caagaaaccc
     3121 gctaagaacc tcaagatgcc cggcgtctac tatgtggaca gaagactgga aagaatcaag
     3181 gaggccgaca aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac
     3241 ctccctagca aactggggca cagataagga tccactagtt ctagacacct gcaacatgcg
     3301 tgagtaaata agcggccgcc accgcggtgg agctctaagc aaatagctaa attatatacg
     3361 aattaatatt atgattaagt gtttacgtga gtgcgatatt tttattacta tcttatacag
     3421 ttgtatatac tctataaaat gagttgtcta ttaattaacg cgatgaatgc tttctgggtt
     3481 tacctctcca acaactctag tttacttctc aatacattca attgtatttg atttgtcaat
     3541 acttcatcat taatcaattc tatagttttg tttttctcgt ttatttccaa atttaatgca
     3601 tcaattttat tattcaattt gtcgttgatt ttggttaatg attttatggt ttgatctctg
     3661 gcattgattg tttgtgttag tttttcatta ttgataatta aattatttaa gttagttatc
     3721 aactcggtgt tttcaagttt caagttttca atttctttag agtttattag atttgtcaaa
     3781 gtttctgaat tgcttgattg gtcctgtaga agagtatttg ttgttgtgga taattgattc
     3841 aatttttgag acaattgctg gaaggcgttg aaatatctag catcaatctc atggtttttt
     3901 tcccgagagt ctcgtagatt caattgtttt aatatatctt gggaccactc ttgatttgaa
     3961 ctcatggaaa ttaaactggg tgttgtgttg tggtgtaatg attgtacccc ctttgcttat
     4021 aattgtgtgg tgacaccgat tatttaaagc tgcagcatac gatatatata catgtgtata
     4081 tatgtatacc tatgaatgtc agtaagtatg tatacgaaca gtatgatact gaagatgaca
     4141 aggtaatgca tcattctata cgtgtcattc tgaacgaggc gcgctttcct tttttctttt
     4201 tgctttttct ttttttttct cttgaactcc agaaaaaaaa tataaaagag atggaggaac
     4261 gggaaaaagt tagttgtggt gataggtggc aagtggtatt ccgtaagaac aacaagaaaa
     4321 gcatttcata ttatggctga actgagcgaa caagtgcaaa atttaagcat caacgacaac
     4381 aacgagaatg gttatgttcc tcctcactta agaggaaaac caagaagtgc cagaaataac
     4441 agtagcaact acaataacaa caacggcggc tacaacggtg gccgtggcgg tggcagcttc
     4501 tttagcaaca accgtcgtgg tggttacggc aacggtggtt tcttcggtgg aaacaacggt
     4561 ggcagcggcc ggccatttaa atgtttaaac ggccacgctg gccccctata gtgagtcgta
     4621 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4681 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4741 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     4801 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4861 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4921 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4981 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5041 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5101 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5161 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5221 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5281 tttcacaccg catagggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     5341 agtattagga tctctaccct tggcgaaaag tcctctgcca acaatgatga tatctgatcc
     5401 accacttaca acttcgtcga cggttctgta ctgctgaccc aatatgcgta tatataccaa
     5461 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     5521 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg
     5581 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5641 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5701 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5761 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5821 atggtttctt agacgtcggc gcgccgagga cgtcaggtgg cacttttcgg ggaaatgtgc
     5881 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5941 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6001 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6061 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6121 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6181 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6241 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6301 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6361 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6421 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6481 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6541 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6601 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6661 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6721 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6781 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6841 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6901 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6961 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7021 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7081 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7141 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7201 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7261 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7321 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7381 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7441 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7501 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7561 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7621 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7681 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7741 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     7801 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     7861 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     7921 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     7981 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt aaccctcact
     8041 aaagggaaca aaa
//
