LOCUS       sequence-469041-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 D262N LCD 
            (186-320).
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 D262N
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             445..636
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(630..652)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    738..877
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     793..812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(892..909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1063..1651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1143..1162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1822..2613,2614..2682))
                     /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     2445..2464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2683..2787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2855..2873
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2900..2905,2906..2922,2923..2928)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(3294..3341)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3337..3355
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    3408..3432
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3845..3865)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(3868..3892)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3902..3919)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3939..3961
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3992..4011)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3993..4011)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4095..4114)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
       61 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
      121 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
      181 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
      241 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
      301 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
      361 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
      421 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
      481 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
      541 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
      601 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
      661 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      721 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
      781 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat
      841 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
      901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1501 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2101 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     2161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     2461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2821 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2881 tcttcaagaa ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat
     2941 cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg
     3001 tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc
     3061 cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc
     3121 tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc
     3181 gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga
     3241 tcatggcgac cacacccgtc ctgtggatat ccggatatag ttcctccttt cagcaaaaaa
     3301 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca
     3361 gcagccaact cagcttcctt tcgggctttg ttagcagcct cgaatcaacc actttgtaca
     3421 agaaagctgg gtcttagaac cgtctgccgg agccgtagct agagctgctg ctagatcccc
     3481 cgtagccgcc ctggtttctg ggcttggcga agtactgtcc ccctccgcca tacggtccgc
     3541 tgcttctgcc gccaaagttt ccgcccttca tggggccgaa gttgctgctc tggttgttgt
     3601 agttgccgaa attgttgtag cttcccccgc ctccgaagtt ggagccatcg ttgccgaagc
     3661 cgttatagcc gtcgccgctt ccgccatagc cgcctccgcc tctagagcct ccaaatccgc
     3721 ccctgccgct gaagttgccg cctctgccaa agttgtcgtt gccgccgaat ccgccgcctc
     3781 ttcctccgcc gaaattgccg ctgccgcttc tgcctctctg gctgctgctg gcgctggcca
     3841 tgctgccctg aaaatacagg ttttcgaagc ctgctttttt gtacaaactt gttgattcga
     3901 ggtgatggtg atggtgatgg tagtacgaca tatgtatatc tccttcttaa agttaaacaa
     3961 aattatttct agagggaaac cgttgtggtc tccctatagt gagtcgtatt aatttcgcgg
     4021 gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     4081 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     4141 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     4201 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     4261 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gatcgaccga
     4321 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     4381 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     4441 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     4501 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     4561 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg
     4621 tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
     4681 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
     4741 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac
     4801 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     4861 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     4921 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
     4981 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
     5041 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
     5101 gtcctggcca cgggtgcgca tgatcgtgct c
//
