LOCUS       sequence-430097-        4548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged N-terminal prion-like domains of Efg1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4548)
  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 4548)
  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-N-PrLD
FEATURES             Location/Qualifiers
     source          1..4548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(358..405)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(965..985)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(986..1003)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    1064..1088
                     /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)."
     primer_bind     complement(1088..1107)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1089..1107)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1571..1588)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1571..1587)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1686..1703)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1857..2445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1937..1956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2531..3613)
                     /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     3575..3594
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             complement(3696..4505)
                     /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"
     promoter        complement(join(4506..4548,1..49))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt
       61 gttacaacca attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc
      121 tcataacacc ccttgtttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
      181 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggactcccc atgcgagagt
      241 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgcc
      301 cgggctaatt agggggtgtc gcccttaggt acgaactcga ttgacgcgtc tcaggggcaa
      361 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctagggggc tcgagttagg
      421 tcggctgctg catggtgcta acatgaggcg gaatcggaac ggcctgctgt ccaggcagcg
      481 gaggctgctg gctcggaccg ctggtgctat tgctcggaat cggtgcacca ctcggtgcgc
      541 tggtataatt acgattgcta ccgttataat gctgaggctg cggctggctc agctgattcg
      601 gaattgtctg ctgatagtaa ttaccctggc tggttgctgc cggatactga tagcggttat
      661 aggtgttata atcatattgc tgctgctgct gttgctgttg ttgctgctga cctgcggtct
      721 gaccggtctg ctgaccaggc tgaccttgct gctgataaaa catatagtta tacggctgct
      781 gaactgctgc cgctgctgcg gttgcggctg cctgttgctg ttgctggctt gcattaccgg
      841 tggtggtcgg ttgctgctgc tgaggaaatg cctgctgatt tgctgcggtg gtctgctgcg
      901 gcataccatt attgtagttg ccgttcatct gattataata cggaatgcta taggtgctca
      961 tatgaccttg aaagtacaga ttttcgtgat ggtggtgatg gtggatcttg tcgctgccca
     1021 tggtatatct ccttcaaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc
     1081 acaattcccc tatagtgagt cgtattaatt tcgcggaagg gcgacacaaa atttattcta
     1141 aatgcataat aaatactgat aacatcttat agtttgtatt atattttgta ttatcgttga
     1201 catgtataat tttgatatca aaaactgatt ttccctttat tattttcgag atttattttc
     1261 ttaattctct ttaacaaact agaaatattg tatatacaaa aaatcataaa taatagatga
     1321 atagtttaat tataggtgtt catcaatcga aaaagcaacg tatcttattt aaagtgcgtt
     1381 gcttttttct catttataag gttaaataat tctcatatat caagcaaagt gacaggcgcc
     1441 cttaaatatt ctgacaaatg ctctttccct aaactccccc cataaaaaaa cccgccgaag
     1501 cgggttttta cgttatttgc ggattaacga ttactcgtta tcagaaccgc ccagggggcc
     1561 cgagcttaag actggccgtc gttttacaac acagaaagag tttgtagaaa cgcaaaaagg
     1621 ccatccgtca ggggccttct gcttagtttg atgcctggca gttccctact ctcgccttcc
     1681 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1741 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1801 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1861 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1921 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1981 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2041 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2101 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2161 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2221 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtgggctaac
     2281 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2341 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2401 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2461 ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt taagggattt
     2521 tggtcatgag tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2581 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     2641 tcaccagtga gactggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     2701 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     2761 gcgggatata acatgagcta tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     2821 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     2881 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     2941 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3001 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3061 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3121 cctcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3181 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     3241 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     3301 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     3361 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     3421 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     3481 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     3541 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     3601 ttactggttt catattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     3661 cgcgaaaggt tttgcgccat tcgatggcgc gccgcttaga aaaactcatc gagcatcaaa
     3721 tgaaattgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     3781 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     3841 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     3901 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     3961 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     4021 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgaggcg aaatacgcga
     4081 tcgctgttaa aaggacaatt acaaacagga atcgagtgca accggcgcag gaacactgcc
     4141 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaacgctgtt
     4201 tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     4261 atggtcggaa gtggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     4321 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     4381 tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     4441 tataaatcag catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg
     4501 ctcatattct tcctttttca atattattga agcatttatc agggttat
//
