LOCUS       sequence-180954-        8078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2 under control of TDH3 constitutive promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8078)
  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 8078)
  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: pHES854
FEATURES             Location/Qualifiers
     source          1..8078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        320..507
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     promoter        1940..2583
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2477..2500
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2596..3294
                     /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(3296..3312)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3296..3312)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4631..4650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4632..4650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4657..4674)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4657..4673)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4666..4688)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4814..5269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4901..4920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5111..5132
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5625..5644)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5744..5766
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5804..5822)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5907..6011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6012..6080,6081..6872)
                     /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(6230..6249)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7043..7631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7532..7551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7785..7802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7919..7940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7955..7960,7961..7978,7979..7985)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7993..8009
                     /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     7998..8020
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8017..8033
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8017..8033
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8054..8072
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aagctgggcc ggccatttaa atgtttaaac ggccacgctg gccgctgcac ggtcctgttc
       61 cctagcatgt acgtgagcgt atttcctttt aaaccacgac gctttgtctt cattcaacgt
      121 ttcccattgt ttttttctac tattgctttg ctgtgggaaa aacttatcga aagatgacga
      181 ctttttctta attctcgttt taagagcttg gtgagcgcta ggagtcactg ccaggtatcg
      241 tttgaacacg gcattagtca gggaagtcat aacacagtcc tttcccgcaa ttttcttttt
      301 ctattactct tggcctcctc tagtacactc tatatttttt tatgcctcgg taatgatttt
      361 catttttttt tttccaccta gcggatgact cttttttttt cttagcgatt ggcattatca
      421 cataatgaat tatacattat ataaagtaat gtgatttctt cgaagaatat actaaaaaat
      481 gagcaggcaa gataaacgaa ggcaaagagg ctctatatgt atataaagct gcggatgcct
      541 tctgatttaa tctagttcta gtttacttca ggtttactta aagttgagat aattgaagca
      601 aagagaagca tcgagcaaag gatggcgttt gttaagaggg ttacgcagga gacgaatata
      661 cagctggcgc tggatcttga cggtgggtct gtttctgtac gggagagcat actgggcaag
      721 gaatatgcta gtggtgatgg gcagaccatc catgtgcaca ctggagttgg gtttttggac
      781 cacatgttga ctgcgctggc gaagcatggc gggtggtctc tgatcctgga gtgtataggg
      841 gacttgcaca tcgatgacca ccacactgtt gaggactgtg ggattgcgct gggccaagcg
      901 ttcaaggagg cgcttggctc cgtgcgtggt atcaagaggt tcgggcatgg gtttgcacca
      961 ctggacgagg cgctgagccg cgctgtggtt gacttctcca ataggccttt cgccgtggtg
     1021 gagctgggcc tgaagcgaga acgcataggc cagctatcca cagagatgat cccgcacttc
     1081 ttggagagtt tcgccactga ggcgcgtatc accatgcatg tggactgtct gcggggcacc
     1141 aacgaccacc accgctccga atcagctttc aaggcgctcg ccatcgccat cagagaggca
     1201 agaacaccta cgggtcgcga tgacgttcca tccactaagg gtgtcctagc atagaacaca
     1261 gcccacagct accaccatca acaatattta tatagataac gtacacatag aaatcacaca
     1321 aacagagtat ttattcttaa ctacatgaac taccatcaga ccgtctgggc cactatataa
     1381 tgtgctattc ataaacgtga tcactctacg tagcaggcga ccccaggtga aaatttttca
     1441 gcgagctgcc agattgtcag gtgaaaaact gaaaaaaact tctgggcgat gagcttgtgg
     1501 cgggaaatta agtatataaa gcagttagtt tcttctgctt cttgtggttc tgggtattct
     1561 tgtaatattt ggaataatag aaaaccaaga ctactgcctt tcttttcata ttatagagga
     1621 agaaatacgc acgaacacga tatagaggta aattttttgc aaaaagatac caaactataa
     1681 aagaaataga agactaaact acaaaaaata aagaaagtaa tacatatatc caatatgact
     1741 gagttagcag agcaagtttc catgcctgaa gggcaacaaa gacgtggtgg ttacgttcca
     1801 cctcacatcc gtaacggtgg tggcagaaga ggcgattctg ctcctcgtgg tggtttcaac
     1861 aacggtggcg gccgtggtgg tttcttcaga aacaacggtg accgtggtgg attcggcttt
     1921 gggtaccggg cccagtttat cattatcaat actcgccatt tcaaagaata cgtaaataat
     1981 taatagtagt gattttccta actttattta gtcaaaaaat tagcctttta attctgctgt
     2041 aacccgtaca tgcccaaaat agggggcggg ttacacagaa tatataacat cgtaggtgtc
     2101 tgggtgaaca gtttattcct ggcatccact aaatataatg gagcccgctt tttaagctgg
     2161 catccagaaa aaaaaagaat cccagcacca aaatattgtt ttcttcacca accatcagtt
     2221 cataggtcca ttctcttagc gcaactacag agaacagggg cacaaacagg caaaaaacgg
     2281 gcacaacctc aatggagtga tgcaacctgc ctggagtaaa tgatgacaca aggcaattga
     2341 cccacgcatg tatctatctc attttcttac accttctatt accttctgct ctctctgatt
     2401 tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga aattattccc ctacttgact
     2461 aataagtata taaagacggt aggtattgat tgtaattctg taaatctatt tcttaaactt
     2521 cttaaattct acttttatag ttagtctttt ttttagtttt aaaacaccag aacttagttt
     2581 cgactcgaga ataaaatggt gagcgagctg attaaggaga acatgcacat gaagctgtac
     2641 atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
     2701 tacgagggca cccagaccat gagaatcaag gcggtcgagg gcggccctct ccccttcgcc
     2761 ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag
     2821 ggcatccccg acttctttaa gcagtccttc cccgagggct tcacatggga gagagtcacc
     2881 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
     2941 ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag
     3001 aagaaaacac tcggctggga ggcctccacc gagaccctgt accccgctga cggcggcctg
     3061 gaaggcagag ccgacatggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttg
     3121 aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat
     3181 gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
     3241 gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacag ataaggatcc
     3301 actagttcta gacacctgca acatgcgtga gtaaataagc ggccgccacc gcggtggagc
     3361 tctaagcaaa tagctaaatt atatacgaat taatattatg attaagtgtt tacgtgagtg
     3421 cgatattttt attactatct tatacagttg tatatactct ataaaatgag ttgtctatta
     3481 attaacgcga tgaatgcttt ctgggtttac ctctccaaca actctagttt acttctcaat
     3541 acattcaatt gtatttgatt tgtcaatact tcatcattaa tcaattctat agttttgttt
     3601 ttctcgttta tttccaaatt taatgcatca attttattat tcaatttgtc gttgattttg
     3661 gttaatgatt ttatggtttg atctctggca ttgattgttt gtgttagttt ttcattattg
     3721 ataattaaat tatttaagtt agttatcaac tcggtgtttt caagtttcaa gttttcaatt
     3781 tctttagagt ttattagatt tgtcaaagtt tctgaattgc ttgattggtc ctgtagaaga
     3841 gtatttgttg ttgtggataa ttgattcaat ttttgagaca attgctggaa ggcgttgaaa
     3901 tatctagcat caatctcatg gtttttttcc cgagagtctc gtagattcaa ttgttttaat
     3961 atatcttggg accactcttg atttgaactc atggaaatta aactgggtgt tgtgttgtgg
     4021 tgtaatgatt gtaccccctt tgcttataat tgtgtggtga caccgattat ttaaagctgc
     4081 agcatacgat atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat
     4141 acgaacagta tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga
     4201 acgaggcgcg ctttcctttt ttctttttgc tttttctttt tttttctctt gaactccaga
     4261 aaaaaaatat aaaagagatg gaggaacggg aaaaagttag ttgtggtgat aggtggcaag
     4321 tggtattccg taagaacaac aagaaaagca tttcatatta tggctgaact gagcgaacaa
     4381 gtgcaaaatt taagcatcaa cgacaacaac gagaatggtt atgttcctcc tcacttaaga
     4441 ggaaaaccaa gaagtgccag aaataacagt agcaactaca ataacaacaa cggcggctac
     4501 aacggtggcc gtggcggtgg cagcttcttt agcaacaacc gtcgtggtgg ttacggcaac
     4561 ggtggtttct tcggtggaaa caacggtggc agcggccggc catttaaatg tttaaacggc
     4621 cacgctggcc ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     4681 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4741 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4801 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4861 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4921 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4981 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5041 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5101 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5161 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5221 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     5281 attttctcct tacgcatctg tgcggtattt cacaccgcat agggtaataa ctgatataat
     5341 taaattgaag ctctaatttg tgagtttagt attaggatct ctacccttgg cgaaaagtcc
     5401 tctgccaaca atgatgatat ctgatccacc acttacaact tcgtcgacgg ttctgtactg
     5461 ctgacccaat atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt
     5521 ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata
     5581 aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat ctgctctgat
     5641 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5701 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5761 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     5821 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcggcgcg ccgaggacgt
     5881 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5941 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6001 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6061 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6121 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6181 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6241 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6301 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6361 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6421 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6481 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6541 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6601 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6661 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6721 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6781 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6841 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6901 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6961 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7021 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7081 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7141 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7201 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7261 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7321 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7381 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7441 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7501 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7561 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7621 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7681 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7741 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     7801 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     7861 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     7921 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7981 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     8041 acgccaagct cggaattaac cctcactaaa gggaacaa
//
