LOCUS       sequence-437750-        5142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged N- and C-terminal prion-like domains of Efg1 
            with Y115/Y132/Y139 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-Y115S/Y132S/Y139S
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             321..1403
                     /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
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDGSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(340..359)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      1489..2077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1978..1997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2231..2248
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2346..2363
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2347..2363
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2827..2846
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2827..2845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2846..2870
                     /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             2931..2948
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2949..2969
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      4123..4170
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4393..4412)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        4479..4570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4571..5142,1..238)
                     /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"
ORIGIN
        1 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
       61 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
      121 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
      181 tgataatcct gatatgaata aattgcaatt tcatttgatg ctcgatgagt ttttctaagc
      241 ggcgcgccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
      301 gtcaattcag ggtggtgaat atgaaaccag taacgttata cgatgtcgca gagtatgccg
      361 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      421 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      481 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      541 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      601 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      661 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      721 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      781 cacccatcaa cagtattatt ttctcccatg aggacggtac gcgactgggc gtggagcatc
      841 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      901 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      961 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     1021 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     1081 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     1141 acgataccga agatggctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     1201 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1261 agggcaatca gctgttgcca gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1321 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1381 cccgactgga aagcgggcag tgactcatga ccaaaatccc ttaacgtgag ttacgcgcgc
     1441 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1501 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1561 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1621 gataccaaat actgttcttc tagtgtagcc gtagttagcc caccacttca agaactctgt
     1681 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1741 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1801 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1861 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1921 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1981 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2041 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2101 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2161 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2221 gaccgagcgc agcgagtcag tgagcgagga agcggaaggc gagagtaggg aactgccagg
     2281 catcaaacta agcagaaggc ccctgacgga tggccttttt gcgtttctac aaactctttc
     2341 tgtgttgtaa aacgacggcc agtcttaagc tcgggccccc tgggcggttc tgataacgag
     2401 taatcgttaa tccgcaaata acgtaaaaac ccgcttcggc gggttttttt atggggggag
     2461 tttagggaaa gagcatttgt cagaatattt aagggcgcct gtcactttgc ttgatatatg
     2521 agaattattt aaccttataa atgagaaaaa agcaacgcac tttaaataag atacgttgct
     2581 ttttcgattg atgaacacct ataattaaac tattcatcta ttatttatga ttttttgtat
     2641 atacaatatt tctagtttgt taaagagaat taagaaaata aatctcgaaa ataataaagg
     2701 gaaaatcagt ttttgatatc aaaattatac atgtcaacga taatacaaaa tataatacaa
     2761 actataagat gttatcagta tttattatgc atttagaata aattttgtgt cgcccttccg
     2821 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     2881 taattttgtt taacttttga aggagatata ccatgggcag cgacaagatc caccatcacc
     2941 accatcacga aaatctgtac tttcaaggtc atatgagcac ctatagcatt ccgtattata
     3001 atcagatgaa cggcaactac aataatggta tgccgcagca gaccaccgca gcaaatcagc
     3061 aggcatttcc tcagcagcag caaccgacca ccaccggtaa tgcaagccag caacagcaac
     3121 aggcagccgc aaccgcagca gcggcagcag ttcagcagcc gtataactat atgttttatc
     3181 agcagcaagg tcagcctggt cagcagaccg gtcagaccgc aggtcagcag caacaacagc
     3241 aacagcagca gcagcaatat gattataaca cctataaccg ctatcagtat ccggcagcaa
     3301 ccagccaggg taattcctat cagcagacaa ttccgaatca gctgagccag ccgcagcctc
     3361 agcattctaa cggtagcaat cgtaattcta ccagcgcacc gagtggtgca ccgattccga
     3421 gcaatagcac cagcggtccg agccagcagc ctccgctgcc tggacagcag gccgttccga
     3481 ttccgcctca tgttagcacc atgcagcagc cgacatcagg taatggtaat agcattagcg
     3541 cgaccagcgg tggtagcaat gttagtggtg caagcggtgc aggtagcacc accagtccgg
     3601 ttaataccaa agcagcgacc gcagccggta ttccgcaggg caactattat cagacctata
     3661 atcagcaaca gtacccgcag cagtatggtc agtataatgc accgggtaaa aatcagaata
     3721 caccggcatc acagcctggt agtaccacca atgatcagta tctgcagcag cagcaacagc
     3781 aacaacaaca aatgtatggt tatcagagca actactatca aggtggtgca gccaatagca
     3841 gctattatcc gaattattac caacaacagc agccgaatta tgcaagcagc tatccgtatc
     3901 aacagcagca acaaaaacag cagcaacaac aaccgaatca gcagcaacag agtgatcagc
     3961 agcagacctc aaccccgagc ggtggtgccg gtacacgtag cgttcatcag agtccgcagg
     4021 ttcagagcct gacccagggt agcgtgcatc cgagcccaca gcagcatcag gcaaatcaga
     4081 gcgcaagcac cgttgcaaaa gaagagaaat aactcgagcc ccctagcata accccttggg
     4141 gcctctaaac gggtcttgag gggttttttg cccctgagac gcgtcaatcg agttcgtacc
     4201 taagggcgac accccctaat tagcccgggc gaaaggccca gtctttcgac tgagcctttc
     4261 gttttatttg atgcctggca gttccctact ctcgcatggg gagtccccac actaccatcg
     4321 gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg
     4381 ccgccaggca aacaaggggt gttatgagcc atattcaggt ataaatgggc tcgcgataat
     4441 gttcagaatt ggttaattgg ttgtaacact gacccctatt tgtttatttt tctaaataca
     4501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4561 aaggaagaat atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     4621 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     4681 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     4741 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     4801 gccacttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     4861 tgcgatcccc ggaaaaacag cgttccaggt attagaagaa tatcctgatt caggtgaaaa
     4921 tattgttgat gcgctggcag tgttcctgcg ccggttgcac tcgattcctg tttgtaattg
     4981 tccttttaac agcgatcgcg tatttcgcct cgctcaggcg caatcacgaa tgaataacgg
     5041 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     5101 gaaagaaatg cataaacttt tgccattctc accggattca gt
//
