LOCUS       sequence-469044-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 LCD 
            (186-320) D262V with partial W mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  AUTHORS   Das T, Zaidi F, Farag M, Ruff KM, Messing J, Taylor JP, Pappu RV, 
            Mittag T
  TITLE     Metastable condensates suppress conversion to amyloid fibrils.
  JOURNAL   bioRxiv [Preprint]. 2024 Mar 3:2024.02.28.582569. doi: 
            10.1101/2024.02.28.582569.
  PUBMED    38464104
REFERENCE   2  (bases 1 to 5131)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Mar 3:2024.02.28.582569. doi: 
            10.1101/2024.02.28.582569."
COMMENT     Sequence Label: A1-LCD 11W D262V
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1138..1157
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1241..1260
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1241..1259
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1291..1313
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1333..1350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    1360..1384
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1387..1407
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1820..1844)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1897..1915)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1911..1958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2324..2329,2330..2346,2347..2352))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2379..2397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2465..2569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2570..2638,2639..3430)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2788..2807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3601..4189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4090..4109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4343..4360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4375..4514
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4440..4459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4600..4622
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4616..4807)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
       61 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
      121 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg
      181 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
      241 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
      301 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
      361 aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc
      421 gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga
      481 gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg
      541 gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc
      601 atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg
      661 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
      721 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
      781 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
      841 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
      901 gagctgactg ggttgaaggc tctcaagggc atcggtcgat cgacgctctc ccttatgcga
      961 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag
     1021 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     1081 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     1141 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     1201 gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg
     1261 agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg agatatacat
     1321 atgtcgtact accatcacca tcaccatcac ctcgaatcaa caagtttgta caaaaaagca
     1381 ggcttcgaga acctgtattt tcagggtagc atggcaagcg caagcagcag ccagcgtggt
     1441 cgtagcggta gcggtaattg gggtggtggt cgcggtggtg gttggggtgg caatgataac
     1501 tggggtcgtg gtggtaattg gagcggtcgc ggaggctggg gtggttcaag aggtggtggc
     1561 ggatggggtg gtagtggtga tggttggaat ggttggggca atgatggtag caattttggt
     1621 ggtggtggca gctataatgt tttcggcaat tataacaatc agagcagcaa ctttggtccg
     1681 atgaaaggcg gtaattttgg cggtcgtagc agcggtccgt atggtggcgg tggtcagtat
     1741 tttgccaaac cgcgtaatca aggtggctgg ggaggtagca gcagcagctc aagctggggt
     1801 agcggtcgtc gttggtaaga cccagctttc ttgtacaaag tggttgattc gaggctgcta
     1861 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     1921 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     1981 gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc
     2041 gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg
     2101 catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg
     2161 tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct
     2221 acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac
     2281 ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg
     2341 ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt
     2401 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     2461 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2521 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2581 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2641 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2701 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2761 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     2821 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2881 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2941 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3001 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3061 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     3121 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3181 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3241 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3301 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3361 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3421 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3481 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3541 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3601 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3661 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3721 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3781 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3841 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3901 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3961 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4021 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4081 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4141 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4201 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4261 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4321 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     4381 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     4441 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     4501 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4561 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4621 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4681 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4741 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4801 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     4861 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     4921 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     4981 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     5041 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     5101 acgaaacacg gaaaccgaag accattcatg t
//
