LOCUS       sequence-430110-        5142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged N- and C-terminal prion-like domains of Efg1 
            with Y401/Y404/Y409 to serine mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5142)
  AUTHORS   Wang SH, Zheng T, Fawzi NL
  TITLE     Structure and interactions of prion-like domains in transcription 
            factor Efg1 phase separation.
  JOURNAL   Biophys J. 2024 Jun 4;123(11):1481-1493. doi: 
            10.1016/j.bpj.2024.01.030. Epub 2024 Feb 1.
  PUBMED    38297837
REFERENCE   2  (bases 1 to 5142)
  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: 
            "10.1016/j.bpj.2024.01.030"; journalName: "Biophys J"; date: 
            "2024-06-4- 4"; volume: "123"; issue: "11"; pages: "1481-1493"
COMMENT     Sequence Label: Efg1-Delta-DBD-Y401S/Y404S/Y409S
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     263..282
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        263..281
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    282..306
                     /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)."
     CDS             367..384
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             385..405
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1559..1606
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1829..1848)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1915..2006
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2007..2816
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2899..3981
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2918..2937)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      4067..4655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4556..4575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4809..4826
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4924..4941
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4925..4941
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aataagatac gttgcttttt cgattgatga acacctataa ttaaactatt catctattat
       61 ttatgatttt ttgtatatac aatatttcta gtttgttaaa gagaattaag aaaataaatc
      121 tcgaaaataa taaagggaaa atcagttttt gatatcaaaa ttatacatgt caacgataat
      181 acaaaatata atacaaacta taagatgtta tcagtattta ttatgcattt agaataaatt
      241 ttgtgtcgcc cttccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
      301 aattcccctc tagaaataat tttgtttaac ttttgaagga gatataccat gggcagcgac
      361 aagatccacc atcaccacca tcacgaaaat ctgtactttc aaggtcatat gagcacctat
      421 agcattccgt attataatca gatgaacggc aactacaata atggtatgcc gcagcagacc
      481 accgcagcaa atcagcaggc atttcctcag cagcagcaac cgaccaccac cggtaatgca
      541 agccagcaac agcaacaggc agccgcaacc gcagcagcgg cagcagttca gcagccgtat
      601 aactatatgt tttatcagca gcaaggtcag cctggtcagc agaccggtca gaccgcaggt
      661 cagcagcaac aacagcaaca gcagcagcag caatatgatt ataacaccta taaccgctat
      721 cagtatccgg cagcaaccag ccagggtaat tactatcagc agacaattcc gaatcagctg
      781 agccagccgc agcctcagca ttataacggt agcaatcgta attataccag cgcaccgagt
      841 ggtgcaccga ttccgagcaa tagcaccagc ggtccgagcc agcagcctcc gctgcctgga
      901 cagcaggccg ttccgattcc gcctcatgtt agcaccatgc agcagccgac atcaggtaat
      961 ggtaatagca ttagcgcgac cagcggtggt agcaatgtta gtggtgcaag cggtgcaggt
     1021 agcaccacca gtccggttaa taccaaagca gcgaccgcag ccggtattcc gcagggcaac
     1081 tattctcaga cctctaatca gcaacagtcc ccgcagcagt atggtcagta taatgcaccg
     1141 ggtaaaaatc agaatacacc ggcatcacag cctggtagta ccaccaatga tcagtatctg
     1201 cagcagcagc aacagcaaca acaacaaatg tatggttatc agagcaacta ctatcaaggt
     1261 ggtgcagcca atagcagcta ttatccgaat tattaccaac aacagcagcc gaattatgca
     1321 agcagctatc cgtatcaaca gcagcaacaa aaacagcagc aacaacaacc gaatcagcag
     1381 caacagagtg atcagcagca gacctcaacc ccgagcggtg gtgccggtac acgtagcgtt
     1441 catcagagtc cgcaggttca gagcctgacc cagggtagcg tgcatccgag cccacagcag
     1501 catcaggcaa atcagagcgc aagcaccgtt gcaaaagaag agaaataact cgagccccct
     1561 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgcccc tgagacgcgt
     1621 caatcgagtt cgtacctaag ggcgacaccc cctaattagc ccgggcgaaa ggcccagtct
     1681 ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggagt
     1741 ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg
     1801 gaccaccgcg ctactgccgc caggcaaaca aggggtgtta tgagccatat tcaggtataa
     1861 atgggctcgc gataatgttc agaattggtt aattggttgt aacactgacc cctatttgtt
     1921 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1981 ttcaataata ttgaaaaagg aagaatatga gccatattca acgggaaacg tcgaggccgc
     2041 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     2101 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     2161 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2221 ggctgacgga atttatgcca cttccgacca tcaagcattt tatccgtact cctgatgatg
     2281 catggttact caccactgcg atccccggaa aaacagcgtt ccaggtatta gaagaatatc
     2341 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcactcga
     2401 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgcctcgct caggcgcaat
     2461 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2521 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2581 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2641 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2701 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2761 ataatcctga tatgaataaa ttgcaatttc atttgatgct cgatgagttt ttctaagcgg
     2821 cgcgccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     2881 caattcaggg tggtgaatat gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     2941 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     3001 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
     3061 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     3121 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     3181 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     3241 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     3301 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     3361 cccatcaaca gtattatttt ctcccatgag gacggtacgc gactgggcgt ggagcatctg
     3421 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
     3481 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
     3541 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
     3601 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
     3661 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
     3721 gataccgaag atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
     3781 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     3841 ggcaatcagc tgttgccagt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     3901 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3961 cgactggaaa gcgggcagtg actcatgacc aaaatccctt aacgtgagtt acgcgcgcgt
     4021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4081 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4141 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4201 taccaaatac tgttcttcta gtgtagccgt agttagccca ccacttcaag aactctgtag
     4261 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4321 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4381 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4441 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4501 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4561 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4621 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4681 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4741 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4801 ccgagcgcag cgagtcagtg agcgaggaag cggaaggcga gagtagggaa ctgccaggca
     4861 tcaaactaag cagaaggccc ctgacggatg gcctttttgc gtttctacaa actctttctg
     4921 tgttgtaaaa cgacggccag tcttaagctc gggccccctg ggcggttctg ataacgagta
     4981 atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt
     5041 tagggaaaga gcatttgtca gaatatttaa gggcgcctgt cactttgctt gatatatgag
     5101 aattatttaa ccttataaat gagaaaaaag caacgcactt ta
//
