LOCUS       sequence-392955-        4293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4293)
  AUTHORS   Molliex A, Temirov J, Lee J, Coughlin M, Kanagaraj AP, Kim HJ, 
            Mittag T, Taylor JP
  TITLE     Phase separation by low complexity domains promotes stress granule 
            assembly and drives pathological fibrillization.
  JOURNAL   Cell. 2015 Sep 24;163(1):123-33. doi: 10.1016/j.cell.2015.09.015.
  PUBMED    26406374
REFERENCE   2  (bases 1 to 4293)
  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.cell.2015.09"; journalName: "Cell"; date: "2015-09-24- 
            24"; volume: "163"; issue: "1"; pages: "123-33"
COMMENT     Sequence Label: His SUMO HNRNPA1 dHexa LCD
FEATURES             Location/Qualifiers
     source          1..4293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..750)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(251..270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(946..2028)
                     /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     1990..2009
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2029..2106)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2152..2171
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2152..2170
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2171..2195
                     /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             2251..2268
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2278..2295
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2308..2601
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             2998..3042
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3080..3098)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3094..3141
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3371..4186)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3371..4186)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4094..4113
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4187..4278)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt
       61 cagctcactc aaaagcggta atacggttat ccacagaatc aggggataaa gccggaaaga
      121 acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt tttccatagg
      181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg
      241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      361 tctcatagct cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccattgg taactgattt
      541 agaggacttt gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa cagattttgg
      601 tgagtgcggt cctccaaccc acttaccttg gttcaaagag ttggtagctc agcgaacctt
      661 gagaaaacca ccgttggtag cggtggtttt tctttattta tgagatgatg aatcaatcgg
      721 tctatcaagt caacgaacag ctattccgtt actctagatt tcagtgcaat ttatctcttc
      781 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
      841 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
      901 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
      961 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1021 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     1081 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     1141 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     1201 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     1261 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     1321 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     1381 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     1441 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     1501 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     1561 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     1621 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     1681 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     1741 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     1801 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     1861 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     1921 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     1981 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     2041 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     2101 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa ttaatacgac
     2161 tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt ttgtttaact
     2221 ttaataagga gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg
     2281 gtgccgcgcg gcagccatat ggctagcatg tcggactcag aagtcaatca agaagctaag
     2341 ccagaggtca agccagaagt caagcctgag actcacatca atttaaaggt gtccgatgga
     2401 tcttcagaga tcttcttcaa gatcaaaaag accactcctt taagaaggct gatggaagcg
     2461 ttcgctaaaa gacagggtaa ggaaatggac tccttaagat tcttgtacga cggtattaga
     2521 attcaagctg atcagacccc tgaagatttg gacatggagg ataacgatat tattgaggct
     2581 cacagagaac agattggtgg atccatggct agtgcttcat ccagccaaag aggtcgaagt
     2641 ggttctggaa actttggtgg tggtcgtgga ggtggtttcg gtgggaatga caacttcggt
     2701 cgtggaggaa acttcagtgg tcgtggtggc tttggtggca gccgtggtgg tggtggatat
     2761 ggtggcagtg gggatggcta taatggattt ggtaatgatg gaagcaattt tggaggtggt
     2821 ggaaattaca acaatcagtc ttcaaatttt ggacccatga agggaggaaa ttttggaggc
     2881 agaagctctg gcccctatgg cggtggaggc caatactttg caaaaccacg aaaccaaggt
     2941 ggctatggcg gttccagcag cagcagtagc tatggcagtg gctaactcga gtctggtaaa
     3001 gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa
     3061 ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     3121 cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg
     3181 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
     3241 tgccctgaac cgacgacaag ctgacgaccg ggtctccgca agtggcactt ttcggggaaa
     3301 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3361 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     3421 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     3481 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     3541 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     3601 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     3661 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     3721 ttcgtgattg cgcctgagcg agacgaaata cgcggtcgct gttaaaagga caattacaaa
     3781 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     3841 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     3901 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     3961 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     4021 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     4081 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     4141 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc
     4201 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4261 tttagaaaaa taaacaaata ggcatgcagc gct
//
