LOCUS       sequence-464532-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 LCD 
            (186-320) D262V with partial W mutant.
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 5W D262V
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     717..736
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     820..839
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        820..838
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             870..892
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             912..929
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    939..963
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             966..986
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1399..1423)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1476..1494)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1490..1537
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1903..1908,1909..1925,1926..1931))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1958..1976)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2044..2148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2149..2217,2218..3009)
                     /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(2367..2386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3180..3768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3669..3688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3922..3939
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3954..4093
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4019..4038)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4179..4201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4195..4386)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
       61 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
      121 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
      181 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
      241 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
      301 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
      361 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
      421 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
      481 agctgactgg gttgaaggct ctcaagggca tcggtcgatc gacgctctcc cttatgcgac
      541 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
      601 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
      661 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
      721 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
      781 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actataggga
      841 gaccacaacg gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata
      901 tgtcgtacta ccatcaccat caccatcacc tcgaatcaac aagtttgtac aaaaaagcag
      961 gcttcgagaa cctgtatttt cagggtagca tggcaagcgc aagcagcagc cagcgtggtc
     1021 gtagcggtag cggtaatttt ggtggtggtc gcggtggtgg ttttggtggc aatgataatt
     1081 ttggccgtgg tggtaattgg agcggtcgtg gtggttgggg tggtagccgt ggcggaggtg
     1141 gctggggagg tagtggtgat ggttataatg gctttggtaa tgatggtagc aactttggtg
     1201 gcggtggtag ctataatgtt tttggcaatt ataacaacca gagcagcaat ttcggtccga
     1261 tgaaaggtgg caattttggc ggtcgtagca gcggtccgta tggcggtggc ggtcagtatt
     1321 ttgccaaacc gcgtaatcaa ggcggttggg gaggttctag cagcagcagc tcatatggta
     1381 gcggtcgtcg ttggtaagac ccagctttct tgtacaaagt ggttgattcg aggctgctaa
     1441 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1501 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1561 atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg
     1621 aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc
     1681 atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt
     1741 cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     1801 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     1861 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     1921 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     1981 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2041 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2101 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2161 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2221 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2281 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2341 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     2401 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2461 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2521 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2581 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2641 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     2701 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2761 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2821 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2881 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2941 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3001 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3061 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3121 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3181 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3241 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3301 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3361 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3421 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3481 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3541 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3601 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3661 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3721 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3781 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3841 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3901 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3961 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     4021 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4081 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4141 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4201 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4261 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4321 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4381 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     4441 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     4501 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     4561 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     4621 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     4681 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     4741 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     4801 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     4861 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     4921 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     4981 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     5041 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     5101 atccatgcca acccgttcca tgtgctcgcc g
//
