LOCUS       sequence-266869-        7308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pRPL18B_FdeR-N_2NLS_tPGK1-pGPM1-fdeO_mcherry_tTDH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7308)
  AUTHORS   Wang R, Cress BF, Yang Z, Hordines JC 3rd, Zhao S, Jung GY, Wang Z, 
            Koffas MAG
  TITLE     Design and Characterization of Biosensors for the Screening of 
            Modular Assembled Naringenin Biosynthetic Library in Saccharomyces 
            cerevisiae.
  JOURNAL   ACS Synth Biol. 2019 Sep 20;8(9):2121-2130. doi: 
            10.1021/acssynbio.9b00212. Epub 2019 Aug 28.
  PUBMED    31433622
REFERENCE   2  (bases 1 to 7308)
  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"; doi: 
            "10.1021/acssynbio.9b00212"; journalName: "ACS Synth Biol"; date: 
            "2019-09-20- 20"; volume: "8"; issue: "9"; pages: "2121-2130"
COMMENT     Sequence Label: S3_005_026_096
FEATURES             Location/Qualifiers
     source          1..7308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(577..597)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(601..621)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     rep_origin      complement(2943..3531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3023..3042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3745..3764)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3756..4571
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3756..4571
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3829..3848)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(5213..6016)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 gttcattatt tcttctttgt tatataatat ccattccata gatgattgcc atccaaacaa
       61 tcaattttac caatctcaaa cccaagtagc gatttggtga gaaccaccct ccctggcaac
      121 agccacaact cgttaccatt gttcatcacg atcatgaaac tcgctgtcag ctgaaatttc
      181 acctcagtgg atctctcttt ttattcttca tcgttccact aacctttttc catcagctgg
      241 cagggaacgg aaagtggaat cccatttagc gagcttcctc ttttcttcaa gaaaagacga
      301 agcttcgttg tagctgtgtt gacatctggt ttggcagcac atagaaatat cgaatgggaa
      361 aaaaaaactg cataaaggca ttaaaagagg agcgaatttt tttttaataa aaatcttaat
      421 aatcattaaa agataaataa tagtctatat atacgtatat aaataaaaaa tattcaaaaa
      481 ataaaataaa ctattatttt agcgtaaagg atggggaaag agaaaagaaa aaaattgatc
      541 tatcgatttc aattcaattc aatgccactc gagttatacc tttctcttct tttttggatc
      601 tacctttctc ttcttttttg gatcggatcc gcagatgcct ggcagcgccg catccatctc
      661 ctgcgcactc tccagaaaca cccgacgcag ccactggatg ccaggatcat tgctgcggta
      721 gcgatgccac tgcatcatct gccgcatctc gcccagcgac agcggactct ccttgatcac
      781 caccggccat tgcggagcca gcagctgcgc cagccgggca tgcaccgtgg cgatgcggtc
      841 cgtcccctgt accagcgcca gcgcagaagc gaagctgaag ctggtcactt ccacccggcg
      901 cgcaaagccc agcttcctgg ccatccacgc ctccaccgac gacgcattgg ccccaggcgg
      961 caccatcacc acatggcctg aggccatgta gcgttccagc gtcagctcgc cttgcgccag
     1021 cgcactgtcg cgccagacca cgcagacatg ccgttcgcgg aagacctctt cggcaggatg
     1081 atcgggcgtg cagaattcct gcggcaagac cagcaggtcc acctcggccc gatccagcga
     1141 gcgggtcggg tcttgcacct gcggcatcag ggcaaagcgg atgtgcttgc cctcggcgtg
     1201 cgcgcgcgcc agcacccggg ggatgaggac ggagagcgta aagtccgaaa ccgagatgcg
     1261 aaactcgcgc gtggactcgg ccggcacgaa ggccggcagc gccgcgatgg agccatcgat
     1321 acgccgcagc acatcatgca ccgcatcctt gagcacctcg gcgcgcggcg tgggctccat
     1381 gcgccggccc acctggatca gcaattcatc atcgaaatac tcgcgcagcc gcgccagggc
     1441 attgctcatg gccgactggc tcagatggat cttttcggcg gcgcggctga tgctcatctc
     1501 cgtgagcagt gcatccaggg cgaccagaag attgaggtcg agcttgttga aacgcataga
     1561 tcttttgttt tttgttttct tctaattgat tttttctttc tatttccttt ggtgaaacag
     1621 gtagttttgt tcaaaagaaa tcattcaatg gtttaaataa aagcaagaag tatgaaaatt
     1681 tttttatttt ttcttcagaa ttttatcata gttgtcgtaa aataacgagg tcgggtaacg
     1741 tgtttggtgt actgaggtct tttttaaaag aagctccagc aagaaagagt tttgtgtgat
     1801 ggctaatgaa gtgtcaattg catttggaaa gcagcctgat ttcttgggcg tgaatatgaa
     1861 attcttcttt tgttatttta taatatgaag tatgaatgaa aacataaagc gcgctgagac
     1921 aacgactgtt caatgtaaaa taaagaaaat ctatcgttct caagacgtgg tcccattttt
     1981 ttttactttg aatttcgtca gcagtgtgac atggttagaa tgtaacccca catttgcaaa
     2041 gagattgtta caggagcgtt ctaattgtcc cgtactggta cattcataag tgaaaagaaa
     2101 gccatatcgg gcgcgctggt ttgtgagcca cagaaaccgg acaaggcact tcttgaatac
     2161 ttcccaagga aagcgccaag gtttacggtt ctgatgggaa tgcagggggg aatctagtct
     2221 tgaagtatcc ttattcctta aaaaaaatat tggacatcct cttcgttcgt ctgcaattat
     2281 cggccagttc agtctagatg cgaattcagg ggtaatgtta tccatgtggg ctttatggac
     2341 cctgaaacca cagccacatt aaccttcttt gatggtcaaa acttatcctt caccataaat
     2401 atgcctcgca aaaaaggtaa ttaacatata tagaattaca ttatttatga aatatcatca
     2461 ctatctctta gcatctttaa tccttttcta catcagataa cttcggtttg ttatcatcgt
     2521 ctgtattgtc atcaattggc gcagtagcct caatttcaac gtcgtttgac tctggtgttt
     2581 gttcatgtgc agatccatga gatgatgaac cgttgccctt ctcaatactc ttcttatcaa
     2641 gtattaattg ctgtagcttt ttttcaatat ctgagtcctc tacaaattta ttgaaaatac
     2701 tcaaaatttt caaaaacgtt tcgttgcttg tcttgcctag tttcaaataa tgcaaataaa
     2761 catttgaaat tattgttaat atggatgaaa tgaggatttt ctgatcttcc tgtgtttcac
     2821 tcgaatttag cattggcggc cgcacggtta tccacagaat caggggataa cgcaggaaag
     2881 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3601 gtcatgactg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt
     3661 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     3721 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     3781 cttgctccag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     3841 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     3901 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     3961 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     4021 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
     4081 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     4141 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgcc
     4201 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     4261 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
     4321 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     4381 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4441 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4501 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     4561 agtttttcta atcagaattg gttaattggt tgtaacacgc ggccgctcgg tctttatatt
     4621 tacatgctaa aaatgggcta caccagagat acataattag atatatatac gccagtacac
     4681 cttatcggcc caagccttgt cccaaggcag cgttttgttc ttggaaacgc tgccctacac
     4741 gttcgctatg cttcaagaac ttttctgagc acttcatgat gcatgtttgt tccttattgg
     4801 ttagctttga tgttgtgaag tcattgacac agtctgtgaa acatctttct accagattag
     4861 agtacaaacg catgaaatcc ttcatttgct tttgttccac tactttttgg aactcttgtt
     4921 gttctttgga gttcaatgcg tccatcttta cagtcctgtc ttattgttct tgatttgtgc
     4981 cccgtaaaat actgttactt ggttctggcg aggtattgga tagttccttt ttataaaggc
     5041 catgaagctt tttctttcca attttttttt tttcgtcatt atagaaatca ttacgaccga
     5101 gattcccggg gcagtggatt caagtgctct actcgggtaa taactgatat aattaaattg
     5161 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt
     5221 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta
     5281 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg
     5341 tagaaaccac atcatccacg gttctatact gttgacccaa tgcatctccc ttgtcatcta
     5401 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc
     5461 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct
     5521 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc
     5581 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc
     5641 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag
     5701 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcctcg aagagtaaaa
     5761 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag
     5821 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact
     5881 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt
     5941 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat
     6001 atgtagcttt cgacatgatt tatcttcgtt tcctgcacgt ttttgttctg tgcagttggg
     6061 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc
     6121 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat
     6181 ttcaaggaaa ccgtgtactg cagtgcacta gtggttcgta acatctctgt aactgctcag
     6241 ccgttcaggg taatatattt taaccgccga cgttctcgcc ataactttta aacagtgata
     6301 actaatattg aaacttagaa agaaaagggt aaagttaatg cttaatcttg tcttggctta
     6361 aaaagtaata tgtacttatt tacgtcatca caaaatctag cagaaaaacg gtagtattta
     6421 tgtatattca aaaaaaaatc attatcctca tcaagattgc tttatctcga gttaggattt
     6481 acttgtacaa ttcatccata ccaccggtag agtgtctacc ttcagctctt tcgtattgtt
     6541 cgacaatggt gtaatcttcg ttgtgagagg taatatccaa cttaatgttg acgttgtaag
     6601 cacctggcaa ttggactggc ttcttagcct tgtaggtggt cttgacttca gcatcgtagt
     6661 gaccaccatc cttcaacttc aatctttgct taatttcacc cttcaaagca ccatcttctg
     6721 ggtacattct ttcagaagaa gcttcccaac ccatggtctt cttttgcatg actggaccat
     6781 cagatgggaa gttggtacct ctcaacttga ccttgtaaat gaattcacca tcttgcaaag
     6841 aagaatcttg ggtgacggtg acgacaccac catcttcgaa gttcatgact ctttcccact
     6901 tgaaaccttc tgggaaagac aacttcaagt aatctggaat atcagctggg tgcttgacgt
     6961 aagccttaga accgtacatg aattgtggag acaaaatatc ccaagcgaat ggcaatggac
     7021 cacccttggt gaccttcaac ttagcggttt gggtaccttc gtatggtcta ccttcacctt
     7081 caccttcaat ttcgaattcg tgaccgttga cagaaccttc catgtggacc ttgaatctca
     7141 tgaattcctt aataatagcc atattgtctt cttcgccctt tgaaaccata gatcttattg
     7201 taatatgtgt gtttgtttgg attattaaga agaataatta caaaaaaaat tacaaaggaa
     7261 ggtaattaca acagaattaa gaaaggacaa gaaggaggaa gagaatca
//
