LOCUS       sequence-422087-        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/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/Q443P
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      27..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     769..786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     884..901
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     885..901
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1365..1384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1365..1383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1384..1408
                     /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             1469..1486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1487..1507
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2661..2708
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2931..2950)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3017..3108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3109..3918
                     /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             4001..5083
                     /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(4020..4039)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
       61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      181 gtgtagccgt agttagccca ccacttcaag aactctgtag caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      781 agcgaggaag cggaaggcga gagtagggaa ctgccaggca tcaaactaag cagaaggccc
      841 ctgacggatg gcctttttgc gtttctacaa actctttctg tgttgtaaaa cgacggccag
      901 tcttaagctc gggccccctg ggcggttctg ataacgagta atcgttaatc cgcaaataac
      961 gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca
     1021 gaatatttaa gggcgcctgt cactttgctt gatatatgag aattatttaa ccttataaat
     1081 gagaaaaaag caacgcactt taaataagat acgttgcttt ttcgattgat gaacacctat
     1141 aattaaacta ttcatctatt atttatgatt ttttgtatat acaatatttc tagtttgtta
     1201 aagagaatta agaaaataaa tctcgaaaat aataaaggga aaatcagttt ttgatatcaa
     1261 aattatacat gtcaacgata atacaaaata taatacaaac tataagatgt tatcagtatt
     1321 tattatgcat ttagaataaa ttttgtgtcg cccttccgcg aaattaatac gactcactat
     1381 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta acttttgaag
     1441 gagatatacc atgggcagcg acaagatcca ccatcaccac catcacgaaa atctgtactt
     1501 tcaaggtcat atgagcacct atagcattcc gtattataat cagatgaacg gcaactacaa
     1561 taatggtatg ccgcagcaga ccaccgcagc aaatcagcag gcatttcctc agcagcagca
     1621 accgaccacc accggtaatg caagccagca acagcaacag cctgccgcaa ccgcacctgc
     1681 ggcagcagtt cagcagccgt ataactatat gttttatccg cagcaaggtc agcctggtca
     1741 gcagaccggt cagaccgcag gtcagcagca acaacagcaa cagcagcagc agcaatatga
     1801 ttataacacc tataaccgct atcagtatcc ggcagcaacc agccagggta attactatca
     1861 gcagacaatt ccgaatcagc tgagccagcc gcagcctcag cattataacg gtagcaatcg
     1921 taattatacc agcgcaccga gtggtgcacc gattccgagc aatagcacca gcggtccgag
     1981 ccagcagcct ccgctgcctg gacagcaggc cgttccgatt ccgcctcatg ttagcaccat
     2041 gcagcagccg acatcaggta atggtaatag cattagcgcg accagcggtg gtagcaatgt
     2101 tagtggtgca agcggtgcag gtagcaccac cagtccggtt aataccaaag cagcgaccgc
     2161 agccggtatt ccgcagggca actattatca gacctataat cagcaacagt acccgcagca
     2221 gtatggtcag tataatgcac cgggtaaaaa tcagaataca ccggcatcac agcctggtag
     2281 taccaccaat gatcagtatc tgcagcagca gccgcagcaa caacaacaaa tgtatggtta
     2341 tcagagcaac tactatcaag gtggtgcagc caatagcagc tattatccga attattacca
     2401 acaacagcag ccgaattatg caagcagcta tccgtatcaa cagcagcaac aaaaacagca
     2461 gcaacaacaa ccgaatcagc agcaacagag tgatcagcag cagacctcaa ccccgagcgg
     2521 tggtgccggt acacgtagcg ttcatcagag tccgcaggtt cagagcctga cccagggtag
     2581 cgtgcatccg agcccacagc agcatcaggc aaatcagagc gcaagcaccg ttgcaaaaga
     2641 agagaaataa ctcgagcccc ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2701 gttttttgcc cctgagacgc gtcaatcgag ttcgtaccta agggcgacac cccctaatta
     2761 gcccgggcga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggcagt
     2821 tccctactct cgcatgggga gtccccacac taccatcggc gctacggcgt ttcacttctg
     2881 agttcggcat ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa caaggggtgt
     2941 tatgagccat attcaggtat aaatgggctc gcgataatgt tcagaattgg ttaattggtt
     3001 gtaacactga cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3061 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatat gagccatatt
     3121 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     3181 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     3241 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     3301 gatgagatgg tcagactaaa ctggctgacg gaatttatgc cacttccgac catcaagcat
     3361 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagcg
     3421 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     3481 ttcctgcgcc ggttgcactc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     3541 tttcgcctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     3601 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     3661 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     3721 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     3781 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     3841 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     3901 ctcgatgagt ttttctaagc ggcgcgccat cgaatggcgc aaaacctttc gcggtatggc
     3961 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat atgaaaccag taacgttata
     4021 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     4081 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     4141 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     4201 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     4261 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     4321 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     4381 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     4441 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aggacggtac
     4501 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     4561 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     4621 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     4681 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     4741 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     4801 tatctcggta gtgggatacg acgataccga agatagctca tgttatatcc cgccgttaac
     4861 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     4921 ctctcagggc caggcggtga agggcaatca gctgttgcca gtctcactgg tgaaaagaaa
     4981 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5041 gcagctggca cgacaggttt cccgactgga aagcgggcag tgactcatga ccaaaatccc
     5101 ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag ac
//
