LOCUS       sequence-430105-        5142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged N- and C-terminal prion-like domains of Efg1 
            with A51/A56/Q70P/N102/Q443 to proline 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-A51P/A56P/Q70P/N102P/Q443P
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..117
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     101..117
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     581..600
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        581..599
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    600..624
                     /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             685..702
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             703..723
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1877..1924
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2147..2166)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2233..2324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2325..3134
                     /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             3217..4299
                     /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(3236..3255)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      4385..4973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4874..4893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(5127..5142,1..2)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aggaagcgga aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga
       61 cggatggcct ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt
      121 aagctcgggc cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa
      181 aaacccgctt cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat
      241 atttaagggc gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga
      301 aaaaagcaac gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt
      361 aaactattca tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga
      421 gaattaagaa aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt
      481 atacatgtca acgataatac aaaatataat acaaactata agatgttatc agtatttatt
      541 atgcatttag aataaatttt gtgtcgccct tccgcgaaat taatacgact cactataggg
      601 gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt ttgaaggaga
      661 tataccatgg gcagcgacaa gatccaccat caccaccatc acgaaaatct gtactttcaa
      721 ggtcatatga gcacctatag cattccgtat tataatcaga tgaacggcaa ctacaataat
      781 ggtatgccgc agcagaccac cgcagcaaat cagcaggcat ttcctcagca gcagcaaccg
      841 accaccaccg gtaatgcaag ccagcaacag caacagcctg ccgcaaccgc acctgcggca
      901 gcagttcagc agccgtataa ctatatgttt tatccgcagc aaggtcagcc tggtcagcag
      961 accggtcaga ccgcaggtca gcagcaacaa cagcaacagc agcagcagca atatgattat
     1021 aacacctatc cgcgctatca gtatccggca gcaaccagcc agggtaatta ctatcagcag
     1081 acaattccga atcagctgag ccagccgcag cctcagcatt ataacggtag caatcgtaat
     1141 tataccagcg caccgagtgg tgcaccgatt ccgagcaata gcaccagcgg tccgagccag
     1201 cagcctccgc tgcctggaca gcaggccgtt ccgattccgc ctcatgttag caccatgcag
     1261 cagccgacat caggtaatgg taatagcatt agcgcgacca gcggtggtag caatgttagt
     1321 ggtgcaagcg gtgcaggtag caccaccagt ccggttaata ccaaagcagc gaccgcagcc
     1381 ggtattccgc agggcaacta ttatcagacc tataatcagc aacagtaccc gcagcagtat
     1441 ggtcagtata atgcaccggg taaaaatcag aatacaccgg catcacagcc tggtagtacc
     1501 accaatgatc agtatctgca gcagcagccg cagcaacaac aacaaatgta tggttatcag
     1561 agcaactact atcaaggtgg tgcagccaat agcagctatt atccgaatta ttaccaacaa
     1621 cagcagccga attatgcaag cagctatccg tatcaacagc agcaacaaaa acagcagcaa
     1681 caacaaccga atcagcagca acagagtgat cagcagcaga cctcaacccc gagcggtggt
     1741 gccggtacac gtagcgttca tcagagtccg caggttcaga gcctgaccca gggtagcgtg
     1801 catccgagcc cacagcagca tcaggcaaat cagagcgcaa gcaccgttgc aaaagaagag
     1861 aaataactcg agccccctag cataacccct tggggcctct aaacgggtct tgaggggttt
     1921 tttgcccctg agacgcgtca atcgagttcg tacctaaggg cgacaccccc taattagccc
     1981 gggcgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     2041 tactctcgca tggggagtcc ccacactacc atcggcgcta cggcgtttca cttctgagtt
     2101 cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaacaag gggtgttatg
     2161 agccatattc aggtataaat gggctcgcga taatgttcag aattggttaa ttggttgtaa
     2221 cactgacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2281 aataaccctg ataaatgctt caataatatt gaaaaaggaa gaatatgagc catattcaac
     2341 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     2401 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     2461 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     2521 agatggtcag actaaactgg ctgacggaat ttatgccact tccgaccatc aagcatttta
     2581 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcgttcc
     2641 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     2701 tgcgccggtt gcactcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2761 gcctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     2821 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     2881 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     2941 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3001 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3061 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcaatttcat ttgatgctcg
     3121 atgagttttt ctaagcggcg cgccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     3181 tagcgcccgg aagagagtca attcagggtg gtgaatatga aaccagtaac gttatacgat
     3241 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     3301 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     3361 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     3421 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     3481 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     3541 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     3601 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     3661 gatgtctctg accagacacc catcaacagt attattttct cccatgagga cggtacgcga
     3721 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     3781 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     3841 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     3901 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     3961 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     4021 tcggtagtgg gatacgacga taccgaagat agctcatgtt atatcccgcc gttaaccacc
     4081 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     4141 cagggccagg cggtgaaggg caatcagctg ttgccagtct cactggtgaa aagaaaaacc
     4201 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4261 ctggcacgac aggtttcccg actggaaagc gggcagtgac tcatgaccaa aatcccttaa
     4321 cgtgagttac gcgcgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4381 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4441 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4501 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttagcccacc
     4561 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4621 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4681 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4741 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4801 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4861 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4921 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4981 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5041 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5101 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cg
//
