LOCUS       sequence-488765-        5097 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  Expresses 6xHis-tagged FUS with LC domain deletion in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5097)
  AUTHORS   Murthy AC, Tang WS, Jovic N, Janke AM, Seo DH, Perdikari TM, Mittal 
            J, Fawzi NL
  TITLE     Molecular interactions contributing to FUS SYGQ LC-RGG phase 
            separation and co-partitioning with RNA polymerase II heptads.
  JOURNAL   Nat Struct Mol Biol. 2021 Nov;28(11):923-935. doi: 
            10.1038/s41594-021-00677-4. Epub 2021 Nov 10.
  PUBMED    34759379
REFERENCE   2  (bases 1 to 5097)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41594-021-00677-4"; journalName: "Nat Struct Mol Biol"; 
            date: "2021-11"; volume: "28"; issue: "11"; pages: "923-935"
COMMENT     Sequence Label: 6xHis-FUS-LCdel
FEATURES             Location/Qualifiers
     source          1..5097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(469..560)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     627..646
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(869..916)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2025..2045)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2046..2063)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2124..2148
                     /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(2148..2167)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2149..2167)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2631..2648)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2631..2647)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2746..2763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2917..3505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2997..3016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3591..4673)
                     /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     4635..4654
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             complement(join(4756..5097,1..468))
                     /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"
ORIGIN
        1 gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca
       61 attacaaaca ggaatcgagt gcaaccggcg caggaacact gccagcgcat caacaatatt
      121 ttcacctgaa tcaggatatt cttctaatac ctggaacgct gtttttccgg ggatcgcagt
      181 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagtggcat
      241 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
      301 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
      361 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
      421 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcatat tcttcctttt
      481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
      541 tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa ttctgaacat
      601 tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgtttg cctggcggca
      661 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
      721 atggtagtgt ggggactccc catgcgagag tagggaactg ccaggcatca aataaaacga
      781 aaggctcagt cgaaagactg ggcctttcgc ccgggctaat tagggggtgt cgcccttagg
      841 tacgaactcg attgacgcgt ctcaggggca aaaaacccct caagacccgt ttagaggccc
      901 caaggggtta tgctaggggg ctcgagtcaa tacggcctct ccctgcgatc ctgtctgtgc
      961 tcacccctgg aatccatctt gccagggcca aagccacctc tgtccccacc accccggccc
     1021 cctcggaagc ctccacggtc cccgccgcgg ccccggtagc cgcctcgatc atagcctcct
     1081 ctgccaccac gacgatcatc cccgtagtta ccccccatgt gagagccacc tggtccccct
     1141 cctgggccat ctggtttagg ggccttacac tggttgcatt cattcctcca agagaagttc
     1201 atattctcac aggtgggatt aggacacttc cagtcaccag ctcgctgctg tcctccaccg
     1261 ccaccacctc cactgggaaa tcctcctcgg ccaccaccac cactgccacc acctccatag
     1321 cctccacggc ccatgggtcc tcctcgccct cggcctccac gaccattgcc accaccccga
     1381 ttaaagtctg cccggcgagt agcaaatgag accttgatag gatttccgga gaattcttta
     1441 ccatcaaacc agtcaatagc tgctttagct gaaggtgggt catcaaaaga gaccgttgcc
     1501 tctcccttca gcttgccagt ttccctgtct gtgtacaaat taatcatggg ctgtcccgtt
     1561 ttcttgtttg tcttaataat accaatctgc ttgaagtaat cagccacaga ctcaattgta
     1621 acattctcac ccaggccttg cacaaagatg gtgttgttgt ctgaattatc ctgttcggag
     1681 tcatgacgtg atccttggtc ccgagggcca ccaaatttat tgaagccacc acggtcactt
     1741 ccgcccatgc cacctctgcc tccacggcca cctccacgac ctctgggttc atagccacca
     1801 ctgctgcggt tgtaaccacc accgccgccg ccgccgccgc caccactgcc acccctgccg
     1861 cggcctccac ggtcctgctg tccatagcca ccgctgccac ctccaccact ctggtcttga
     1921 ttgccataac cgccaccact gccaccacca ctactcatgg aggattgatc ttggccatag
     1981 ttacctccac ctccacctcc acctccacca ccactgctca tatggccttg aaagtataag
     2041 ttctcatgat gatggtggtg atgaatcttg tcgctaccca tggtatatct ccttcaaaag
     2101 ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt
     2161 cgtattaatt tcgcggaagg gcgacacaaa atttattcta aatgcataat aaatactgat
     2221 aacatcttat agtttgtatt atattttgta ttatcgttga catgtataat tttgatatca
     2281 aaaactgatt ttccctttat tattttcgag atttattttc ttaattctct ttaacaaact
     2341 agaaatattg tatatacaaa aaatcataaa taatagatga atagtttaat tataggtgtt
     2401 catcaatcga aaaagcaacg tatcttattt aaagtgcgtt gcttttttct catttataag
     2461 gttaaataat tctcatatat caagcaaagt gacaggcgcc cttaaatatt ctgacaaatg
     2521 ctctttccct aaactccccc cataaaaaaa cccgccgaag cgggttttta cgttatttgc
     2581 ggattaacga ttactcgtta tcagaaccgc ccagggggcc cgagcttaag actggccgtc
     2641 gttttacaac acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca ggggccttct
     2701 gcttagtttg atgcctggca gttccctact ctcgccttcc gcttcctcgc tcactgactc
     2761 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3301 tgtaggcggt gctacagagt tcttgaagtg gtgggctaac tacggctaca ctagaagaac
     3361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3541 tcagtggaac gacgcgcgcg taactcacgt taagggattt tggtcatgag tcactgcccg
     3601 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3661 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gactggcaac
     3721 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     3781 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagcta
     3841 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     3901 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     3961 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4021 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4081 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4141 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cctcatggga gaaaataata
     4201 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4261 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4321 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4381 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4441 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     4501 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     4561 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     4621 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt catattcacc
     4681 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4741 tcgatggcgc gccgcttaga aaaactcatc gagcatcaaa tgaaattgca atttattcat
     4801 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     4861 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     4921 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     4981 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     5041 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaacc
//
