LOCUS       sequence-430112-        5142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged N- and C-terminal prion-like domains of Efg1 
            with Y478/Y482/Y484 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-Y478S/Y482S/Y484S
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(254..273)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        340..431
                     /gene="bla"
                     /label=AmpR promoter
     CDS             432..1241
                     /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             1324..2406
                     /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(1343..1362)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      2492..3080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2981..3000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3234..3251
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3349..3366
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3350..3366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3830..3849
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3830..3848
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3849..3873
                     /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             3934..3951
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3952..3972
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      join(5126..5142,1..31)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ggcctctaaa cgggtcttga ggggtttttt gcccctgaga cgcgtcaatc gagttcgtac
       61 ctaagggcga caccccctaa ttagcccggg cgaaaggccc agtctttcga ctgagccttt
      121 cgttttattt gatgcctggc agttccctac tctcgcatgg ggagtcccca cactaccatc
      181 ggcgctacgg cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact
      241 gccgccaggc aaacaagggg tgttatgagc catattcagg tataaatggg ctcgcgataa
      301 tgttcagaat tggttaattg gttgtaacac tgacccctat ttgtttattt ttctaaatac
      361 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      421 aaaggaagaa tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
      481 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
      541 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
      601 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
      661 tgccacttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
      721 ctgcgatccc cggaaaaaca gcgttccagg tattagaaga atatcctgat tcaggtgaaa
      781 atattgttga tgcgctggca gtgttcctgc gccggttgca ctcgattcct gtttgtaatt
      841 gtccttttaa cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga atgaataacg
      901 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
      961 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     1021 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     1081 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     1141 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     1201 ataaattgca atttcatttg atgctcgatg agtttttcta agcggcgcgc catcgaatgg
     1261 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     1321 aatatgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     1381 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     1441 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     1501 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     1561 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     1621 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     1681 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     1741 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     1801 attttctccc atgaggacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     1861 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     1921 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     1981 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     2041 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     2101 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagatagc
     2161 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     2221 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     2281 ccagtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     2341 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2401 cagtgactca tgaccaaaat cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt
     2461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2641 ttctagtgta gccgtagtta gcccaccact tcaagaactc tgtagcaccg cctacatacc
     2701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3121 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3181 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3241 cagtgagcga ggaagcggaa ggcgagagta gggaactgcc aggcatcaaa ctaagcagaa
     3301 ggcccctgac ggatggcctt tttgcgtttc tacaaactct ttctgtgttg taaaacgacg
     3361 gccagtctta agctcgggcc ccctgggcgg ttctgataac gagtaatcgt taatccgcaa
     3421 ataacgtaaa aacccgcttc ggcgggtttt tttatggggg gagtttaggg aaagagcatt
     3481 tgtcagaata tttaagggcg cctgtcactt tgcttgatat atgagaatta tttaacctta
     3541 taaatgagaa aaaagcaacg cactttaaat aagatacgtt gctttttcga ttgatgaaca
     3601 cctataatta aactattcat ctattattta tgattttttg tatatacaat atttctagtt
     3661 tgttaaagag aattaagaaa ataaatctcg aaaataataa agggaaaatc agtttttgat
     3721 atcaaaatta tacatgtcaa cgataataca aaatataata caaactataa gatgttatca
     3781 gtatttatta tgcatttaga ataaattttg tgtcgccctt ccgcgaaatt aatacgactc
     3841 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     3901 tgaaggagat ataccatggg cagcgacaag atccaccatc accaccatca cgaaaatctg
     3961 tactttcaag gtcatatgag cacctatagc attccgtatt ataatcagat gaacggcaac
     4021 tacaataatg gtatgccgca gcagaccacc gcagcaaatc agcaggcatt tcctcagcag
     4081 cagcaaccga ccaccaccgg taatgcaagc cagcaacagc aacaggcagc cgcaaccgca
     4141 gcagcggcag cagttcagca gccgtataac tatatgtttt atcagcagca aggtcagcct
     4201 ggtcagcaga ccggtcagac cgcaggtcag cagcaacaac agcaacagca gcagcagcaa
     4261 tatgattata acacctataa ccgctatcag tatccggcag caaccagcca gggtaattac
     4321 tatcagcaga caattccgaa tcagctgagc cagccgcagc ctcagcatta taacggtagc
     4381 aatcgtaatt ataccagcgc accgagtggt gcaccgattc cgagcaatag caccagcggt
     4441 ccgagccagc agcctccgct gcctggacag caggccgttc cgattccgcc tcatgttagc
     4501 accatgcagc agccgacatc aggtaatggt aatagcatta gcgcgaccag cggtggtagc
     4561 aatgttagtg gtgcaagcgg tgcaggtagc accaccagtc cggttaatac caaagcagcg
     4621 accgcagccg gtattccgca gggcaactat tatcagacct ataatcagca acagtacccg
     4681 cagcagtatg gtcagtataa tgcaccgggt aaaaatcaga atacaccggc atcacagcct
     4741 ggtagtacca ccaatgatca gtatctgcag cagcagcaac agcaacaaca acaaatgtat
     4801 ggttatcaga gcaactacta tcaaggtggt gcagccaata gcagctatta tccgaattat
     4861 taccaacaac agcagccgaa ttctgcaagc agctctccgt ctcaacagca gcaacaaaaa
     4921 cagcagcaac aacaaccgaa tcagcagcaa cagagtgatc agcagcagac ctcaaccccg
     4981 agcggtggtg ccggtacacg tagcgttcat cagagtccgc aggttcagag cctgacccag
     5041 ggtagcgtgc atccgagccc acagcagcat caggcaaatc agagcgcaag caccgttgca
     5101 aaagaagaga aataactcga gccccctagc ataacccctt gg
//
