LOCUS       sequence-469043-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 LCD 
            (186-320) D262V with all aromatic amino acids mutated to W keeping 
            the hexapeptide fibril core (SYNDFG) intact..
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 allW D262V
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(358..1149,1150..1218))
                     /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     981..1000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1219..1323)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1391..1409
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1436..1441,1442..1458,1459..1464)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(1830..1877)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1873..1891
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    1944..1968
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2381..2401)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2404..2428)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2438..2455)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2475..2497
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2528..2547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2529..2547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2631..2650)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             4112..4303
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4297..4319)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4405..4544
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4460..4479
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4559..4576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4730..5131,1..187))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4810..4829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
       61 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      121 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      181 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      301 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      361 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      421 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      481 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      541 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      601 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      661 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      721 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      781 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      841 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      901 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      961 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1021 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1081 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1141 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1201 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1321 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1381 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct catgtttgac
     1441 agcttatcat cgataagctt taatgcggta gtttatcaca gttaaattgc taacgcagtc
     1501 aggcaccgtg tatgaaatct aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg
     1561 atgctgtagg cataggcttg gttatgccgg tactgccggg cctcttgcgg gatatcgtcc
     1621 attccgacag catcgccagt cactatggcg tgctgctagc gctatatgcg ttgatgcaat
     1681 ttctatgcgc acccgttctc ggagcactgt ccgaccgctt tggccgccgc ccagtcctgc
     1741 tcgcttcgct acttggagcc actatcgact acgcgatcat ggcgaccaca cccgtcctgt
     1801 ggatatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc
     1861 ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg
     1921 gctttgttag cagcctcgaa tcaaccactt tgtacaagaa agctgggtct taccaacgac
     1981 gacctgaacc ccagctgctg ctgctagagc tacctcccca accgccttga ttacgcggtt
     2041 ttgcccacca ctgaccgcca cctccccacg gaccgctgct acgacctccc cagttaccac
     2101 ctttcatcgg accccaattg ctgctctgat tattccaatt gccaaagaca ttatagctgc
     2161 caccgccacc ccagttgcta ccatcattgc cccaaccatt ccaaccatca ccactaccac
     2221 cccatccgcc accacctctt gaaccacccc agcctccgcg accgctccaa ttaccaccac
     2281 gaccccagtt atcattgcca ccccaaccac caccgcgacc accaccccaa ttaccgctac
     2341 cgctacgacc acgctggctg ctgcttgcgc ttgccatgct accctgaaaa tacaggttct
     2401 cgaagcctgc ttttttgtac aaacttgttg attcgaggtg atggtgatgg tgatggtagt
     2461 acgacatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag ggaaaccgtt
     2521 gtggtctccc tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc
     2581 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     2641 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     2701 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
     2761 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     2821 gcaggagtcg cataagggag agcgtcgatc gaccgatgcc cttgagagcc ttcaacccag
     2881 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2941 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     3001 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     3061 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     3121 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     3181 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     3241 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     3301 tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg gcgatttatg
     3361 ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg
     3421 tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag
     3481 ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg
     3541 gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc
     3601 cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat
     3661 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     3721 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     3781 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     3841 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     3901 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     3961 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     4021 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     4081 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     4141 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     4201 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     4261 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4321 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4381 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4441 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt
     4501 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     4561 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4621 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4681 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4741 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5101 gcagagcgag gtatgtaggc ggtgctacag a
//
