LOCUS       sequence-246054-        5583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression for affinity purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5583)
  AUTHORS   Murthy AC, Dignon GL, Kan Y, Zerze GH, Parekh SH, Mittal J, Fawzi NL
  TITLE     Molecular interactions underlying liquid-liquid phase separation of 
            the FUS low-complexity domain.
  JOURNAL   Nat Struct Mol Biol. 2019 Jul;26(7):637-648. doi: 
            10.1038/s41594-019-0250-x. Epub 2019 Jul 1.
  PUBMED    31270472
REFERENCE   2  (bases 1 to 5583)
  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: 
            "10.1038/s41594-019-0250-x"; journalName: "Nat Struct Mol Biol"; 
            date: "2019-07"; volume: "26"; issue: "7"; pages: "637-648"
COMMENT     Sequence Label: pJ411 FUS 1-163 Y24xF
FEATURES             Location/Qualifiers
     source          1..5583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     208..227
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        208..226
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    227..251
                     /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             312..329
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             330..350
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             731..742
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      1945..1992
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2215..2234)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2301..2392
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2393..3202
                     /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             3285..4367
                     /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(3304..3323)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      4453..5041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4942..4961
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5195..5212
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5310..5327
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5311..5327
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attatttatg attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat
       61 aaatctcgaa aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg
      121 ataatacaaa atataataca aactataaga tgttatcagt atttattatg catttagaat
      181 aaattttgtg tcgcccttcc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      241 ataacaattc ccctctagaa ataattttgt ttaacttttg aaggagatat accatgggct
      301 ctgataaaat tcatcaccat caccatcacg aaaatcttta ttttcaaggt atggcctcaa
      361 acgattttac ccaacaagca acccaaagct ttggggcctt ccccacccag cccgggcagg
      421 gcttttccca gcagagcagt cagcccttcg gacagcagag tttcagtggt tttagccagt
      481 ccacggacac ttcaggcttt ggccagagca gcttttcttc ttttggccag agccagaaca
      541 caggctttgg aactcagtca actccccagg gatttggctc gactggcggc tttggcagta
      601 gccagagctc ccaatcgtct ttcgggcagc agtcctcctt tcctggcttt ggccagcagc
      661 cagctcccag cagcacctcg ggaagtttcg gtagcagttc tcagagcagc agctttgggc
      721 agccccagag tgggagcttc agccagcagc ctagctttgg tggacagcag caaagctttg
      781 gacagcagca aagctttaat ccccctcagg gctttggaca gcagaaccag ttcaacagct
      841 aaagcagtgg tggtggaggt ggaggtggag gtggaggtaa ctatggccaa gatcaatcct
      901 ccatgagtag tggtggtggc agtggtggcg gttatggcaa tcaagaccag agtggtggag
      961 gtggcagcgg tggctatgga cagcaggacc gtggaggccg cggcaggggt ggcagtggtg
     1021 gcggcggcgg cggcggcggt ggtggttaca accgcagcag tggtggctat gaacccagag
     1081 gtcgtggagg tggccgtgga ggcagaggtg gcatgggcgg aagtgaccgt ggtggcttca
     1141 ataaatttgg tggccctcgg gaccaaggat cacgtcatga ctccgaacag gataattcag
     1201 acaacaacac catctttgtg caaggcctgg gtgagaatgt tacaattgag tctgtggctg
     1261 attacttcaa gcagattggt attattaaga caaacaagaa aacgggacag cccatgatta
     1321 atttgtacac agacagggaa actggcaagc tgaagggaga ggcaacggtc tcttttgatg
     1381 acccaccttc agctaaagca gctattgact ggtttgatgg taaagaattc tccggaaatc
     1441 ctatcaaggt ctcatttgct actcgccggg cagactttaa tcggggtggt ggcaatggtc
     1501 gtggaggccg agggcgagga ggacccatgg gccgtggagg ctatggaggt ggtggcagtg
     1561 gtggtggtgg ccgaggagga tttcccagtg gaggtggtgg cggtggagga cagcagcgag
     1621 ctggtgactg gaagtgtcct aatcccacct gtgagaatat gaacttctct tggaggaatg
     1681 aatgcaacca gtgtaaggcc cctaaaccag atggcccagg agggggacca ggtggctctc
     1741 acatgggggg taactacggg gatgatcgtc gtggtggcag aggaggctat gatcgaggcg
     1801 gctaccgggg ccgcggcggg gaccgtggag gcttccgagg gggccggggt ggtggggaca
     1861 gaggtggctt tggccctggc aagatggatt ccaggggtga gcacagacag gatcgcaggg
     1921 agaggccata ttgactcgag ccccctagca taaccccttg gggcctctaa acgggtcttg
     1981 aggggttttt tgcccctgag acgcgtcaat cgagttcgta cctaagggcg acacccccta
     2041 attagcccgg gcgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg
     2101 cagttcccta ctctcgcatg gggagtcccc acactaccat cggcgctacg gcgtttcact
     2161 tctgagttcg gcatggggtc aggtgggacc accgcgctac tgccgccagg caaacaaggg
     2221 gtgttatgag ccatattcag gtataaatgg gctcgcgata atgttcagaa ttggttaatt
     2281 ggttgtaaca ctgaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2341 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga atatgagcca
     2401 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2461 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2521 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2581 tacagatgag atggtcagac taaactggct gacggaattt atgccacttc cgaccatcaa
     2641 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2701 agcgttccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2761 agtgttcctg cgccggttgc actcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2821 cgtatttcgc ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2881 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2941 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3001 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3061 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3121 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc aatttcattt
     3181 gatgctcgat gagtttttct aagcggcgcg ccatcgaatg gcgcaaaacc tttcgcggta
     3241 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatatgaaa ccagtaacgt
     3301 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     3361 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     3421 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     3481 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     3541 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     3601 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     3661 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     3721 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaggacg
     3781 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     3841 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     3901 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     3961 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     4021 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     4081 cggatatctc ggtagtggga tacgacgata ccgaagatag ctcatgttat atcccgccgt
     4141 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     4201 aactctctca gggccaggcg gtgaagggca atcagctgtt gccagtctca ctggtgaaaa
     4261 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4321 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgactc atgaccaaaa
     4381 tcccttaacg tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     4441 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4501 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4561 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4621 agcccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4681 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4741 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4801 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4861 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4921 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4981 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5041 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5101 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5161 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5221 aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga cggatggcct
     5281 ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc
     5341 cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa aaacccgctt
     5401 cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat atttaagggc
     5461 gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga aaaaagcaac
     5521 gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt aaactattca
     5581 tct
//
