LOCUS       sequence-469036-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 LCD 
            (186-320) with 3 Gly and 1 Ser mutated to Asn.
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 -3G1S+4V
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4..51
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(417..422,423..439,440..445))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(472..490)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        558..662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(663..731,732..1523)
                     /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(881..900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1694..2282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2183..2202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2436..2453
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2468..2607
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2533..2552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2693..2715
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2709..2900)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4362..4381
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4465..4484
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4465..4483
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4515..4537
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4557..4574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4584..4608
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4611..4631
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(5044..5068)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(join(5121..5131,1..8))
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
       61 ggaactatat ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt
      121 ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc
      181 gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg
      241 actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac
      301 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
      361 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
      421 aagcttatcg atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga
      481 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
      541 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      601 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      661 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      721 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      781 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      841 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      901 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      961 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1021 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1081 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1141 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1201 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1261 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1321 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1381 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1441 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1501 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1561 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1621 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1681 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1741 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1801 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     1861 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1921 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1981 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2041 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2101 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2161 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2221 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2281 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2341 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2401 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2461 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     2521 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     2581 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     2641 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2701 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     2761 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     2821 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     2881 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     2941 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     3001 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     3061 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     3121 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     3181 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     3241 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     3301 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     3361 acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg
     3421 gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat
     3481 tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccattc
     3541 aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata
     3601 gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg
     3661 ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg agcgatcctt
     3721 gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca
     3781 tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg
     3841 cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct
     3901 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     3961 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     4021 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     4081 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     4141 aggctctcaa gggcatcggt cgatcgacgc tctcccttat gcgactcctg cattaggaag
     4201 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     4261 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     4321 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     4381 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     4441 atcgagatct cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc
     4501 cctctagaaa taattttgtt taactttaag aaggagatat acatatgtcg tactaccatc
     4561 accatcacca tcacctcgaa tcaacaagtt tgtacaaaaa agcaggcttc gagaacctgt
     4621 attttcaggg tagcatggca agcgcaagca gcagccagcg tggtcgtagc ggtagcggta
     4681 attttggtgt tggtcgtggt ggtggttttg gtggtaatga taattttggc cgtggtggca
     4741 attttagcgg tcgcggtggc tttggcggtg ttcgtggtgg cggtggctat ggtggcagcg
     4801 gtgatggtta taatggtttt ggcaatgatg gtagcaactt tggaggtggt ggtagctata
     4861 acgattttgg caactataat aaccagagca gcaatttcgt tccgatgaaa ggtggtaact
     4921 ttggtggtcg ttcaagcggt ccgtatggcg gtggtggtca gtattttgcc aaaccgcgta
     4981 atcaggttgg ttatggtggt tctagcagca gcagctcata tggtagcggt cgtcgttttt
     5041 aagacccagc tttcttgtac aaagtggttg attcgaggct gctaacaaag cccgaaagga
     5101 agctgagttg gctgctgcca ccgctgagca a
//
