LOCUS       sequence-469038-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 LCD 
            (186-320) 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
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             199..390
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(384..406)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    492..631
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     547..566
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(646..663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(817..1405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(897..916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1576..2367,2368..2436))
                     /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     2199..2218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2437..2541)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2609..2627
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2654..2659,2660..2676,2677..2682)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(3048..3095)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3091..3109
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    3162..3186
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3599..3619)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(3622..3646)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3656..3673)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3693..3715
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3746..3765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3747..3765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3849..3868)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
       61 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
      121 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
      181 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
      241 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
      301 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
      361 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
      421 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      481 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
      541 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
      601 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
      661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1681 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1861 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     1921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2161 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     2221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2461 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2521 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2581 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     2641 agaattctca tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt
     2701 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
     2761 tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc
     2821 tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc
     2881 tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg
     2941 gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg
     3001 cgaccacacc cgtcctgtgg atatccggat atagttcctc ctttcagcaa aaaacccctc
     3061 aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc
     3121 aactcagctt cctttcgggc tttgttagca gcctcgaatc aaccactttg tacaagaaag
     3181 ctgggtctta ccaacgacga cctgaacccc agctgctgct gctagagcta cctccccaac
     3241 cgccttgatt acgcggtttt gcccaccact gaccgccacc tccccacgga ccgctgctac
     3301 gacctcccca gttaccacct ttcatcggac cccaattgct gctctgatta ttccaattgc
     3361 caaagtcatt atagctgcca ccgccacccc agttgctacc atcattgccc caaccattcc
     3421 aaccatcacc actaccaccc catccgccac cacctcttga accaccccag cctccgcgac
     3481 cgctccaatt accaccacga ccccagttat cattgccacc ccaaccacca ccgcgaccac
     3541 caccccaatt accgctaccg ctacgaccac gctggctgct gcttgcgctt gccatgctac
     3601 cctgaaaata caggttctcg aagcctgctt ttttgtacaa acttgttgat tcgaggtgat
     3661 ggtgatggtg atggtagtac gacatatgta tatctccttc ttaaagttaa acaaaattat
     3721 ttctagaggg aaaccgttgt ggtctcccta tagtgagtcg tattaatttc gcgggatcga
     3781 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     3841 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     3901 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     3961 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     4021 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgatcga ccgatgccct
     4081 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     4141 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     4201 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     4261 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     4321 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
     4381 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     4441 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     4501 gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga
     4561 tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag
     4621 gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca
     4681 cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg
     4741 agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc
     4801 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg
     4861 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
     4921 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
     4981 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
     5041 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
     5101 ctgctggcta ccctgtggaa cacctacatc t
//
