LOCUS       sequence-430101-        5142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged N- and C-terminal prion-like domains of Efg1 
            with A51/A56 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
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      83..671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     572..591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     825..842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     940..957
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     941..957
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1421..1440
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1421..1439
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1440..1464
                     /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             1525..1542
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1543..1563
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2717..2764
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2987..3006)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3073..3164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3165..3974
                     /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             4057..5139
                     /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(4076..4095)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg
       61 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
      121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      241 agccgtagtt agcccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      721 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      781 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      841 aggaagcgga aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga
      901 cggatggcct ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt
      961 aagctcgggc cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa
     1021 aaacccgctt cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat
     1081 atttaagggc gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga
     1141 aaaaagcaac gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt
     1201 aaactattca tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga
     1261 gaattaagaa aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt
     1321 atacatgtca acgataatac aaaatataat acaaactata agatgttatc agtatttatt
     1381 atgcatttag aataaatttt gtgtcgccct tccgcgaaat taatacgact cactataggg
     1441 gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt ttgaaggaga
     1501 tataccatgg gcagcgacaa gatccaccat caccaccatc acgaaaatct gtactttcaa
     1561 ggtcatatga gcacctatag cattccgtat tataatcaga tgaacggcaa ctacaataat
     1621 ggtatgccgc agcagaccac cgcagcaaat cagcaggcat ttcctcagca gcagcaaccg
     1681 accaccaccg gtaatgcaag ccagcaacag caacagcctg ccgcaaccgc acctgcggca
     1741 gcagttcagc agccgtataa ctatatgttt tatcagcagc aaggtcagcc tggtcagcag
     1801 accggtcaga ccgcaggtca gcagcaacaa cagcaacagc agcagcagca atatgattat
     1861 aacacctata accgctatca gtatccggca gcaaccagcc agggtaatta ctatcagcag
     1921 acaattccga atcagctgag ccagccgcag cctcagcatt ataacggtag caatcgtaat
     1981 tataccagcg caccgagtgg tgcaccgatt ccgagcaata gcaccagcgg tccgagccag
     2041 cagcctccgc tgcctggaca gcaggccgtt ccgattccgc ctcatgttag caccatgcag
     2101 cagccgacat caggtaatgg taatagcatt agcgcgacca gcggtggtag caatgttagt
     2161 ggtgcaagcg gtgcaggtag caccaccagt ccggttaata ccaaagcagc gaccgcagcc
     2221 ggtattccgc agggcaacta ttatcagacc tataatcagc aacagtaccc gcagcagtat
     2281 ggtcagtata atgcaccggg taaaaatcag aatacaccgg catcacagcc tggtagtacc
     2341 accaatgatc agtatctgca gcagcagcaa cagcaacaac aacaaatgta tggttatcag
     2401 agcaactact atcaaggtgg tgcagccaat agcagctatt atccgaatta ttaccaacaa
     2461 cagcagccga attatgcaag cagctatccg tatcaacagc agcaacaaaa acagcagcaa
     2521 caacaaccga atcagcagca acagagtgat cagcagcaga cctcaacccc gagcggtggt
     2581 gccggtacac gtagcgttca tcagagtccg caggttcaga gcctgaccca gggtagcgtg
     2641 catccgagcc cacagcagca tcaggcaaat cagagcgcaa gcaccgttgc aaaagaagag
     2701 aaataactcg agccccctag cataacccct tggggcctct aaacgggtct tgaggggttt
     2761 tttgcccctg agacgcgtca atcgagttcg tacctaaggg cgacaccccc taattagccc
     2821 gggcgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     2881 tactctcgca tggggagtcc ccacactacc atcggcgcta cggcgtttca cttctgagtt
     2941 cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaacaag gggtgttatg
     3001 agccatattc aggtataaat gggctcgcga taatgttcag aattggttaa ttggttgtaa
     3061 cactgacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3121 aataaccctg ataaatgctt caataatatt gaaaaaggaa gaatatgagc catattcaac
     3181 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     3241 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     3301 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     3361 agatggtcag actaaactgg ctgacggaat ttatgccact tccgaccatc aagcatttta
     3421 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcgttcc
     3481 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     3541 tgcgccggtt gcactcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     3601 gcctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     3661 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     3721 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3781 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3841 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3901 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcaatttcat ttgatgctcg
     3961 atgagttttt ctaagcggcg cgccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
     4021 tagcgcccgg aagagagtca attcagggtg gtgaatatga aaccagtaac gttatacgat
     4081 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     4141 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     4201 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     4261 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     4321 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     4381 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     4441 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     4501 gatgtctctg accagacacc catcaacagt attattttct cccatgagga cggtacgcga
     4561 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     4621 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     4681 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     4741 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     4801 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     4861 tcggtagtgg gatacgacga taccgaagat agctcatgtt atatcccgcc gttaaccacc
     4921 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     4981 cagggccagg cggtgaaggg caatcagctg ttgccagtct cactggtgaa aagaaaaacc
     5041 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5101 ctggcacgac aggtttcccg actggaaagc gggcagtgac tc
//
