LOCUS       sequence-266891-        7366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pYTK096-pRPL18B_ NLS_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 7366)
  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 7366)
  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_008_026_096
FEATURES             Location/Qualifiers
     source          1..7366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(137..157)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(161..181)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1152..1172)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     rep_origin      complement(2561..3149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2641..2660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3363..3382)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3374..4189
                     /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             3374..4189
                     /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(3447..3466)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(4831..5634)
                     /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 tagtctatat atacgtatat aaataaaaaa tattcaaaaa ataaaataaa ctattatttt
       61 agcgtaaagg atggggaaag agaaaagaaa aaaattgatc tatcgatttc aattcaattc
      121 aatgccactc gagttatacc tttctcttct tttttggatc tacctttctc ttcttttttg
      181 gatcggatcc gcagatgcct ggcagcctgg tctcaggatc cgcagatgcc tggcagcgcc
      241 gcatccatct cctgcgcact ctccagaaac acccgacgca gccactggat gccaggatca
      301 ttgctgcggt agcgatgcca ctgcatcatc tgccgcatct cgcccagcga cagcggactc
      361 tccttgatca ccaccggcca ttgcggagcc agcagctgcg ccagccgggc atgcaccgtg
      421 gcgatgcggt ccgtcccctg taccagcgcc agcgcagaag cgaagctgaa gctggtcact
      481 tccacccggc gcgcaaagcc cagcttcctg gccatccacg cctccaccga cgacgcattg
      541 gccccaggcg gcaccatcac cacatggcct gaggccatgt agcgttccag cgtcagctcg
      601 ccttgcgcca gcgcactgtc gcgccagacc acgcagacat gccgttcgcg gaagacctct
      661 tcggcaggat gatcgggcgt gcagaattcc tgcggcaaga ccagcaggtc cacctcggcc
      721 cgatccagcg agcgggtcgg gtcttgcacc tgcggcatca gggcaaagcg gatgtgcttg
      781 ccctcggcgt gcgcgcgcgc cagcacccgg gggatgagga cggagagcgt aaagtccgaa
      841 accgagatgc gaaactcgcg cgtggactcg gccggcacga aggccggcag cgccgcgatg
      901 gagccatcga tacgccgcag cacatcatgc accgcatcct tgagcacctc ggcgcgcggc
      961 gtgggctcca tgcgccggcc cacctggatc agcaattcat catcgaaata ctcgcgcagc
     1021 cgcgccaggg cattgctcat ggccgactgg ctcagatgga tcttttcggc ggcgcggctg
     1081 atgctcatct ccgtgagcag tgcatccagg gcgaccagaa gattgaggtc gagcttgttg
     1141 aaacgagaac caacctttct cttcttcttt ggcatagatc ttttgttttt tgttttcttc
     1201 taattgattt tttctttcta tttcctttgg tgaaacaggt agttttgttc aaaagaaatc
     1261 attcaatggt ttaaataaaa gcaagaagta tgaaaatttt tttatttttt cttcagaatt
     1321 ttatcatagt tgtcgtaaaa taacgaggtc gggtaacgtg tttggtgtac tgaggtcttt
     1381 tttaaaagaa gctccagcaa gaaagagttt tgtgtgatgg ctaatgaagt gtcaattgca
     1441 tttggaaagc agcctgattt cttgggcgtg aatatgaaat tcttcttttg ttattttata
     1501 atatgaagta tgaatgaaaa cataaagcgc gctgagacaa cgactgttca atgtaaaata
     1561 aagaaaatct atcgttctca agacgtggtc ccattttttt ttactttgaa tttcgtcagc
     1621 agtgtgacat ggttagaatg taaccccaca tttgcaaaga gattgttaca ggagcgttct
     1681 aattgtcccg tactggtaca ttcataagtg aaaagaaagc catatcgggc gcgctggttt
     1741 gtgagccaca gaaaccggac aaggcacttc ttgaatactt cccaaggaaa gcgccaaggt
     1801 ttacggttct gatgggaatg caggggggaa tctagtcttg aagtatcctt attccttaaa
     1861 aaaaatattg gacatcctct tcgttcgtct gcaattatcg gccagttcag tctagatgcg
     1921 aattcagggg taatgttatc catgtgggct ttatggaccc tgaaaccaca gccacattaa
     1981 ccttctttga tggtcaaaac ttatccttca ccataaatat gcctcgcaaa aaaggtaatt
     2041 aacatatata gaattacatt atttatgaaa tatcatcact atctcttagc atctttaatc
     2101 cttttctaca tcagataact tcggtttgtt atcatcgtct gtattgtcat caattggcgc
     2161 agtagcctca atttcaacgt cgtttgactc tggtgtttgt tcatgtgcag atccatgaga
     2221 tgatgaaccg ttgcccttct caatactctt cttatcaagt attaattgct gtagcttttt
     2281 ttcaatatct gagtcctcta caaatttatt gaaaatactc aaaattttca aaaacgtttc
     2341 gttgcttgtc ttgcctagtt tcaaataatg caaataaaca tttgaaatta ttgttaatat
     2401 ggatgaaatg aggattttct gatcttcctg tgtttcactc gaatttagca ttggcggccg
     2461 cacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2521 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3001 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgactgat ccttcaactc
     3241 agcaaaagtt cgatttattc aacaaagcca cgttgtgtct caaaatctct gatgttacat
     3301 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     3361 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctccaggc cgcgattaaa
     3421 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     3481 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     3541 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     3601 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3661 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     3721 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3781 ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat
     3841 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3901 acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca
     3961 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     4021 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     4081 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     4141 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     4201 taattggttg taacacgcgg ccgctcggtc tttatattta catgctaaaa atgggctaca
     4261 ccagagatac ataattagat atatatacgc cagtacacct tatcggccca agccttgtcc
     4321 caaggcagcg ttttgttctt ggaaacgctg ccctacacgt tcgctatgct tcaagaactt
     4381 ttctgagcac ttcatgatgc atgtttgttc cttattggtt agctttgatg ttgtgaagtc
     4441 attgacacag tctgtgaaac atctttctac cagattagag tacaaacgca tgaaatcctt
     4501 catttgcttt tgttccacta ctttttggaa ctcttgttgt tctttggagt tcaatgcgtc
     4561 catctttaca gtcctgtctt attgttcttg atttgtgccc cgtaaaatac tgttacttgg
     4621 ttctggcgag gtattggata gttccttttt ataaaggcca tgaagctttt tctttccaat
     4681 tttttttttt tcgtcattat agaaatcatt acgaccgaga ttcccggggc agtggattca
     4741 agtgctctac tcgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     4801 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
     4861 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
     4921 aaatagtcct cttccaacaa taataatgtc agatcctgta gaaaccacat catccacggt
     4981 tctatactgt tgacccaatg catctccctt gtcatctaaa cccacaccgg gtgtcataat
     5041 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
     5101 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     5161 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     5221 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     5281 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     5341 accaatgtca gcaaattttc tgtcctcgaa gagtaaaaaa ttgtacttgg cggataatgc
     5401 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
     5461 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     5521 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     5581 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     5641 tcttcgtttc ctgcacgttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     5701 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     5761 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gtgtactgca
     5821 gtgcactagt ggttcgtaac atctctgtaa ctgctcagcc gttcagggta atatatttta
     5881 accgccgacg ttctcgccat aacttttaaa cagtgataac taatattgaa acttagaaag
     5941 aaaagggtaa agttaatgct taatcttgtc ttggcttaaa aagtaatatg tacttattta
     6001 cgtcatcaca aaatctagca gaaaaacggt agtatttatg tatattcaaa aaaaaatcat
     6061 tatcctcatc aagattgctt tatctcgagt taggatttac ttgtacaatt catccatacc
     6121 accggtagag tgtctacctt cagctctttc gtattgttcg acaatggtgt aatcttcgtt
     6181 gtgagaggta atatccaact taatgttgac gttgtaagca cctggcaatt ggactggctt
     6241 cttagccttg taggtggtct tgacttcagc atcgtagtga ccaccatcct tcaacttcaa
     6301 tctttgctta atttcaccct tcaaagcacc atcttctggg tacattcttt cagaagaagc
     6361 ttcccaaccc atggtcttct tttgcatgac tggaccatca gatgggaagt tggtacctct
     6421 caacttgacc ttgtaaatga attcaccatc ttgcaaagaa gaatcttggg tgacggtgac
     6481 gacaccacca tcttcgaagt tcatgactct ttcccacttg aaaccttctg ggaaagacaa
     6541 cttcaagtaa tctggaatat cagctgggtg cttgacgtaa gccttagaac cgtacatgaa
     6601 ttgtggagac aaaatatccc aagcgaatgg caatggacca cccttggtga ccttcaactt
     6661 agcggtttgg gtaccttcgt atggtctacc ttcaccttca ccttcaattt cgaattcgtg
     6721 accgttgaca gaaccttcca tgtggacctt gaatctcatg aattccttaa taatagccat
     6781 attgtcttct tcgccctttg aaaccataga tcttattgta atatgtgtgt ttgtttggat
     6841 tattaagaag aataattaca aaaaaaatta caaaggaagg taattacaac agaattaaga
     6901 aaggacaaga aggaggaaga gaatcagttc attatttctt ctttgttata taatatccat
     6961 tccatagatg attgccatcc aaacaatcaa ttttaccaat ctcaaaccca agtagcgatt
     7021 tggtgagaac caccctccct ggcaacagcc acaactcgtt accattgttc atcacgatca
     7081 tgaaactcgc tgtcagctga aatttcacct cagtggatct ctctttttat tcttcatcgt
     7141 tccactaacc tttttccatc agctggcagg gaacggaaag tggaatccca tttagcgagc
     7201 ttcctctttt cttcaagaaa agacgaagct tcgttgtagc tgtgttgaca tctggtttgg
     7261 cagcacatag aaatatcgaa tgggaaaaaa aaactgcata aaggcattaa aagaggagcg
     7321 aatttttttt taataaaaat cttaataatc attaaaagat aaataa
//
