LOCUS       sequence-349922-        8312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SED1 biosensor for expression in Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8312)
  AUTHORS   Cuevas-Velazquez CL, Vellosillo T, Guadalupe K, Schmidt HB, Yu F, 
            Moses D, Brophy JAN, Cosio-Acosta D, Das A, Wang L, Jones AM, 
            Covarrubias AA, Sukenik S, Dinneny JR
  TITLE     Intrinsically disordered protein biosensor tracks the 
            physical-chemical effects of osmotic stress on cells.
  JOURNAL   Nat Commun. 2021 Sep 14;12(1):5438. doi: 10.1038/s41467-021-25736-8.
  PUBMED    34521831
REFERENCE   2  (bases 1 to 8312)
  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: "Nat 
            Commun."; date: "2021-09-14"; volume: "12(1):5438. doi"; pages: " 
            10.1038/s41467-021-25736-8"
COMMENT     Sequence Label: pDRFLIP38-AtLEA4-5
FEATURES             Location/Qualifiers
     source          1..8312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /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     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2510..2521
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2939..2955)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2940..2956)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2980..2999)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2981..2999)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3147..3334
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3202..3226)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3346..3363)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3346..3362)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3355..3377)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3798..5144
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5255..6058)
                     /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"
     promoter        complement(6059..6274)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6299..6317)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6385..6489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6490..6558,6559..7350)
                     /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(6708..6727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7521..8109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8010..8029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8263..8280
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgacc atgattacgc caagcttcct gaaacggaga aacataaaca
      241 ggcattgctg ggatcaccca tacatcactc tgttttgcct gaccttttcc ggtaatttga
      301 aaacaaaccc ggtctcgaag cggagatccg gcgataatta ccgcagaaat aaacccatac
      361 acgagacgta gaaccagccg cacatggccg gagaaactcc tgcgagaatt tcgtaaactc
      421 gcgcgcattg catctgtatt tcctaatgcg gcacttccag gcctcgatcg agaccgttta
      481 tccattgctt ttttgttgtc tttttccctc gttcacagaa agtctgaaga agctatagta
      541 gaactatgag ctttttttgt ttctgttttc cttttttttt tttttacctc tgtggaaatt
      601 gttactctca cactctttag ttcgtttgtt tgttttgttt attccaatta tgaccggtga
      661 cgaaacgtgg tcgatggtgg gtaccgctta tgctcccctc cattagtttc gattatataa
      721 aaaggccaaa tattgtatta ttttcaaatg tcctatcatt atcgtctaac atctaatttc
      781 tcttaaattt tttctctttc tttcctataa caccaatagt gaaaatcttt ttttcttcta
      841 tatctacaaa aacttttttt ttctatcaac ctcgttgata aattttttct ttaacaatcg
      901 ttaataatta attaattgga aaataaccat tttttctctc ttttatacac acattcaaaa
      961 gaaagaaaaa aaatataccc cagcctcgat ctagaatggt gagcaaaggt gaagaactgt
     1021 ttaccggtgt tgttccgatt ctggttgaac tggatggtga cgttaatggt cacaaatttt
     1081 cagttagcgg tgaaggcgaa ggtgatgcaa cctatggtaa actgaccctg aaatttatct
     1141 gtaccaccgg caaactgccg gttccgtggc cgaccctggt taccaccctg agctggggtg
     1201 ttcagtgttt tgcacgttat ccggatcaca tgaaacagca cgattttttc aaaagcgcaa
     1261 tgccggaagg ttatgttcaa gaacgtacca tcttcttcaa agatgacggc aactataaaa
     1321 cccgtgccga agttaaattt gaaggtgata ccctggtgaa tcgcattgaa ctgaaaggca
     1381 tcgattttaa agaggatggt aatatcctgg gccacaaact ggaatataat gccattcatg
     1441 gcaacgtgta tatcaccgca gataaacaga aaaacggcat caaagcaaat tttggcctga
     1501 actgcaatat tgaagatggt tcagttcagc tggcagatca ttatcagcag aataccccga
     1561 ttggtgatgg tccggttctg ctgccggata atcattatct gagcacccag agcaaactga
     1621 gcaaagatcc gaatgaaaaa cgtgatcaca tggtgctgct ggaatttgtt accgcagcag
     1681 gtattaccct gggtatggat gaactgtata aagagctcat gcagtcgatg aaagaaacag
     1741 cttcgaatat tgcagcttct gcaaaatctg gcatggacaa aaccaaagct accttggagg
     1801 aaaaggcgga gaagatgaag acacgagacc ctgttcagaa acagatggct acacaggtta
     1861 aagaagataa gatcaatcaa gctgagatgc agaagagaga aacgcgtcag cacaacgcgg
     1921 ccatgaaaga agcggctgga gccggaaccg gtttaggttt ggggacggcc actcactcga
     1981 ccactggaca agtcggacac ggcactggga cacatcagat gtcggctctg cctggtcacg
     2041 gaacgggaca actgaccgac cgcgttgtgg agggcacggc tgtgaccgac ccgattggaa
     2101 ggaacactgg aactggtcgg acaaccgctc ataacacaca cgttggtggt ggtggtgcca
     2161 ccgggtacgg aaccggcggg ggatatactg gaagatctat ggtgtcaaaa ggtgaggaac
     2221 tgtttacagg cgtggtgccg atcctggtag agctggacgg ggatgtgaat ggccataaat
     2281 tcagcgtttc aggtgaaggt gagggcgacg ccacgtacgg aaaactgaca ctgaaattca
     2341 tttgcacaac aggtaaactg cctgtgcctt ggcctacact ggtgaccacc tttggttatg
     2401 gtctgatgtg ctttgctcgc tatcctgacc acatgaaaca acatgatttc tttaaatctg
     2461 ccatgcctga aggctacgtg caagagcgca ccattttttt caaagacgat gggaattaca
     2521 aaacacgtgc ggaggtgaaa ttcgagggcg atacactggt taaccgtatc gagctgaaag
     2581 gtatcgactt caaagaggac ggaaacattc tgggtcataa actggaatac aactacaaca
     2641 gccataacgt gtacatcatg gccgacaaac aaaaaaacgg gattaaagtg aacttcaaaa
     2701 tccgccacaa catcgaagat ggcagcgtgc agctggccga ccactatcaa caaaacacac
     2761 cgatcggcga cggtcctgta ctgctgcctg acaaccacta tctgtcatat cagagcgcac
     2821 tgtcaaaaga tcctaacgag aaacgcgacc acatggttct gctggaattc gtgacagccg
     2881 ctggcattac actgggcatg gacgagctgt acaaataaga attcgatatc aagcttatcg
     2941 ataccgtcga cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac
     3001 gcgcggatcc agctttggac ttcttcgcca gaggtttggt caagtctcca atcaaggttg
     3061 tcggcttgtc taccttgcca gaaatttacg aaaagatgga aaagggtcaa atcgttggta
     3121 gatacgttgt tgacacttct aaataagcga atttcttatg atttatgatt tttattatta
     3181 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     3241 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc
     3301 atgaggtcgc tcttattgac cacacctcta ccggcatgcc aattcactgg ccgtcgtttt
     3361 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3421 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3481 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3541 tatttcacac cgcataatcg gatcgtactt gttacccatc attgaatttt gaacatccga
     3601 acctgggagt tttccctgaa acagatagta tatttgaacc tgtataataa tatatagtct
     3661 agcgctttac ggaagacaat gtatgtattt cggttcctgg agaaactatt gcatctattg
     3721 cataggtaat cttgcacgtc gcatccccgg ttcattttct gcgtttccat cttgcacttc
     3781 aatagcatat ctttgttaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     3841 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     3901 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     3961 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4021 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     4081 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     4141 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     4201 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     4261 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     4321 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     4381 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     4441 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     4501 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     4561 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     4621 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     4681 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt
     4741 ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata ggaacttcgg
     4801 aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg
     4861 cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta tatatatata
     4921 catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat gcgtctattt
     4981 atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca tgcggggtat
     5041 cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc ctcaattgga
     5101 ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcgatccg atgataagct
     5161 gtcaaacatg agaattgggt aataactgat ataattaaat tgaagctcta atttgtgagt
     5221 ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca
     5281 aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct
     5341 ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca
     5401 cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca
     5461 taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga
     5521 taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag
     5581 atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta
     5641 cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat
     5701 tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg
     5761 tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata
     5821 atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg
     5881 tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg
     5941 tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct
     6001 tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga
     6061 tttatcttcg tttcctgcat gtttttgttc tgtgcagttg ggttaagaat actgggcaat
     6121 ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc
     6181 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc
     6241 aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagcta attcttgaag acgaaagggc
     6301 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6361 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6421 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6481 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6541 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6601 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6661 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6721 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6781 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6841 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6901 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6961 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7021 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7081 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7141 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7201 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7261 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7321 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7381 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7441 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7501 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7561 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7621 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7681 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7741 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7801 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7861 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7921 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7981 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8041 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8101 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8161 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     8221 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     8281 gaagcggaag agcgcccaat acgcaaaccg cc
//
