LOCUS       sequence-430107-        5142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged N- and C-terminal prion-like domains of Efg1 
            with Y64/Y66/Y69 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-Y64S/Y66S/Y69S
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     467..486
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        467..485
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    486..510
                     /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             571..588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             589..609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1763..1810
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2033..2052)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2119..2210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2211..3020
                     /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             3103..4185
                     /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(3122..3141)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      4271..4859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4760..4779
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5013..5030
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     join(5128..5142,1..3)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(5129..5142,1..3)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agtcttaagc tcgggccccc tgggcggttc tgataacgag taatcgttaa tccgcaaata
       61 acgtaaaaac ccgcttcggc gggttttttt atggggggag tttagggaaa gagcatttgt
      121 cagaatattt aagggcgcct gtcactttgc ttgatatatg agaattattt aaccttataa
      181 atgagaaaaa agcaacgcac tttaaataag atacgttgct ttttcgattg atgaacacct
      241 ataattaaac tattcatcta ttatttatga ttttttgtat atacaatatt tctagtttgt
      301 taaagagaat taagaaaata aatctcgaaa ataataaagg gaaaatcagt ttttgatatc
      361 aaaattatac atgtcaacga taatacaaaa tataatacaa actataagat gttatcagta
      421 tttattatgc atttagaata aattttgtgt cgcccttccg cgaaattaat acgactcact
      481 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taacttttga
      541 aggagatata ccatgggcag cgacaagatc caccatcacc accatcacga aaatctgtac
      601 tttcaaggtc atatgagcac ctatagcatt ccgtattata atcagatgaa cggcaactac
      661 aataatggta tgccgcagca gaccaccgca gcaaatcagc aggcatttcc tcagcagcag
      721 caaccgacca ccaccggtaa tgcaagccag caacagcaac aggcagccgc aaccgcagca
      781 gcggcagcag ttcagcagcc gtctaactct atgttttctc agcagcaagg tcagcctggt
      841 cagcagaccg gtcagaccgc aggtcagcag caacaacagc aacagcagca gcagcaatat
      901 gattataaca cctataaccg ctatcagtat ccggcagcaa ccagccaggg taattactat
      961 cagcagacaa ttccgaatca gctgagccag ccgcagcctc agcattataa cggtagcaat
     1021 cgtaattata ccagcgcacc gagtggtgca ccgattccga gcaatagcac cagcggtccg
     1081 agccagcagc ctccgctgcc tggacagcag gccgttccga ttccgcctca tgttagcacc
     1141 atgcagcagc cgacatcagg taatggtaat agcattagcg cgaccagcgg tggtagcaat
     1201 gttagtggtg caagcggtgc aggtagcacc accagtccgg ttaataccaa agcagcgacc
     1261 gcagccggta ttccgcaggg caactattat cagacctata atcagcaaca gtacccgcag
     1321 cagtatggtc agtataatgc accgggtaaa aatcagaata caccggcatc acagcctggt
     1381 agtaccacca atgatcagta tctgcagcag cagcaacagc aacaacaaca aatgtatggt
     1441 tatcagagca actactatca aggtggtgca gccaatagca gctattatcc gaattattac
     1501 caacaacagc agccgaatta tgcaagcagc tatccgtatc aacagcagca acaaaaacag
     1561 cagcaacaac aaccgaatca gcagcaacag agtgatcagc agcagacctc aaccccgagc
     1621 ggtggtgccg gtacacgtag cgttcatcag agtccgcagg ttcagagcct gacccagggt
     1681 agcgtgcatc cgagcccaca gcagcatcag gcaaatcaga gcgcaagcac cgttgcaaaa
     1741 gaagagaaat aactcgagcc ccctagcata accccttggg gcctctaaac gggtcttgag
     1801 gggttttttg cccctgagac gcgtcaatcg agttcgtacc taagggcgac accccctaat
     1861 tagcccgggc gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca
     1921 gttccctact ctcgcatggg gagtccccac actaccatcg gcgctacggc gtttcacttc
     1981 tgagttcggc atggggtcag gtgggaccac cgcgctactg ccgccaggca aacaaggggt
     2041 gttatgagcc atattcaggt ataaatgggc tcgcgataat gttcagaatt ggttaattgg
     2101 ttgtaacact gacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2161 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat atgagccata
     2221 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2281 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     2341 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2401 cagatgagat ggtcagacta aactggctga cggaatttat gccacttccg accatcaagc
     2461 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     2521 cgttccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2581 tgttcctgcg ccggttgcac tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2641 tatttcgcct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2701 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2761 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2821 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2881 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2941 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcaa tttcatttga
     3001 tgctcgatga gtttttctaa gcggcgcgcc atcgaatggc gcaaaacctt tcgcggtatg
     3061 gcatgatagc gcccggaaga gagtcaattc agggtggtga atatgaaacc agtaacgtta
     3121 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     3181 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     3241 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     3301 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     3361 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     3421 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     3481 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     3541 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaggacggt
     3601 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     3661 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     3721 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     3781 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     3841 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     3901 gatatctcgg tagtgggata cgacgatacc gaagatagct catgttatat cccgccgtta
     3961 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     4021 ctctctcagg gccaggcggt gaagggcaat cagctgttgc cagtctcact ggtgaaaaga
     4081 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4141 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgactcat gaccaaaatc
     4201 ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4261 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4321 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4381 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4441 cccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4501 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4561 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4621 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4681 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4741 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4801 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4861 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4921 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4981 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5041 gcgagagtag ggaactgcca ggcatcaaac taagcagaag gcccctgacg gatggccttt
     5101 ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg cc
//
