LOCUS       sequence-469028-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 LCD 
            (186-320) D262N 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 D262N
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             128..319
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(313..335)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    421..560
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     476..495
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(575..592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(746..1334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(826..845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1505..2296,2297..2365))
                     /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     2128..2147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2366..2470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2538..2556
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2583..2588,2589..2605,2606..2611)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2977..3024)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3020..3038
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    3091..3115
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3528..3548)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(3551..3575)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3585..3602)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3622..3644
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3675..3694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3676..3694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3778..3797)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
       61 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
      121 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
      181 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
      241 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
      301 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      361 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      421 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
      481 tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac agatgcgtaa
      541 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      601 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      661 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      721 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      781 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      841 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      901 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      961 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1021 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1081 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1141 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1201 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1261 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1321 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1381 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1441 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1501 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1561 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1621 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1681 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1741 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1801 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1861 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1921 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1981 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2041 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2101 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     2161 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2221 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2281 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2341 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2401 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2461 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     2521 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctcat
     2581 gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt aaattgctaa
     2641 cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc
     2701 accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct cttgcgggat
     2761 atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct atatgcgttg
     2821 atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg ccgccgccca
     2881 gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc gaccacaccc
     2941 gtcctgtgga tatccggata tagttcctcc tttcagcaaa aaacccctca agacccgttt
     3001 agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc
     3061 ctttcgggct ttgttagcag cctcgaatca accactttgt acaagaaagc tgggtcttac
     3121 caacgacgac ctgaacccca gctgctgctg ctagagctac ctccccaacc gccttgatta
     3181 cgcggttttg cccaccactg accgccacct ccccacggac cgctgctacg acctccccag
     3241 ttaccacctt tcatcggacc ccaattgctg ctctgattat tccaattgcc aaagttatta
     3301 tagctgccac cgccacccca gttgctacca tcattgcccc aaccattcca accatcacca
     3361 ctaccacccc atccgccacc acctcttgaa ccaccccagc ctccgcgacc gctccaatta
     3421 ccaccacgac cccagttatc attgccaccc caaccaccac cgcgaccacc accccaatta
     3481 ccgctaccgc tacgaccacg ctggctgctg cttgcgcttg ccatgctacc ctgaaaatac
     3541 aggttctcga agcctgcttt tttgtacaaa cttgttgatt cgaggtgatg gtgatggtga
     3601 tggtagtacg acatatgtat atctccttct taaagttaaa caaaattatt tctagaggga
     3661 aaccgttgtg gtctccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc
     3721 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
     3781 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     3841 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
     3901 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     3961 tcctaatgca ggagtcgcat aagggagagc gtcgatcgac cgatgccctt gagagccttc
     4021 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     4081 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
     4141 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
     4201 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
     4261 caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg
     4321 acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg
     4381 cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa
     4441 ggatcgctcg cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg
     4501 atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta
     4561 taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga
     4621 atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat
     4681 tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg
     4741 ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc
     4801 gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt
     4861 agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg
     4921 cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc
     4981 ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac
     5041 cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc
     5101 tctggtcccg ccgcatccat accgccagtt g
//
