LOCUS       sequence-469035-        5131 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged hnRNPA1 LCD 
            (186-320) with 4 aspartates mutated to valine.
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 Apr 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 -4D+4V
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(239..244,245..261,262..267))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(294..312)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        380..484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(485..553,554..1345)
                     /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(703..722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1516..2104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2005..2024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2258..2275
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2290..2429
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2355..2374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2515..2537
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2531..2722)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4184..4203
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4287..4306
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4287..4305
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4337..4359
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4379..4396
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4406..4430
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4433..4453
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4866..4890)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4943..4961)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4957..5004
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag
       61 tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata
      121 accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg
      181 ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat
      241 taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt
      301 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      361 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      421 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      481 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      541 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      601 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      661 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      721 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      781 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      841 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      901 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      961 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1021 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1081 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1141 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1201 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1261 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1321 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1381 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1441 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1621 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2221 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2281 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     2341 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     2401 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     2461 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2521 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     2581 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     2641 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     2701 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     2761 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     2821 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     2881 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     2941 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     3001 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     3061 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     3121 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     3181 gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg
     3241 cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga
     3301 attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat
     3361 tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta
     3421 tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat
     3481 cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc
     3541 ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg
     3601 catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt
     3661 cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg
     3721 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     3781 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     3841 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     3901 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     3961 gaaggctctc aagggcatcg gtcgatcgac gctctccctt atgcgactcc tgcattagga
     4021 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     4081 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     4141 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     4201 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     4261 ggatcgagat ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
     4321 tccctctaga aataattttg tttaacttta agaaggagat atacatatgt cgtactacca
     4381 tcaccatcac catcacctcg aatcaacaag tttgtacaaa aaagcaggct tcgagaacct
     4441 gtattttcag ggtagcatgg caagcgcaag cagcagccag cgtggtcgta gcggtagcgg
     4501 taattttggt ggtggtcgcg gtggtggttt tggtggcaat gtgaattttg gccgtggtgg
     4561 taattttagc ggtcgtggtg gctttggcgg tagccgtggc ggtggcggtt atggcggtag
     4621 tggtgttggt tataatggtt ttggcaatgt tggtagcaac tttggaggtg gtggtagcta
     4681 taatgtgttc ggcaattata acaatcagag cagcaatttc ggtccgatga aaggtggcaa
     4741 ttttggcgga cgtagcagcg gtccgtatgg tggtggcggt cagtattttg ccaaaccgcg
     4801 taatcaaggt ggctatggtg gttctagcag cagcagctca tatggtagtg gtcgtcgttt
     4861 ttaagaccca gctttcttgt acaaagtggt tgattcgagg ctgctaacaa agcccgaaag
     4921 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     4981 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac
     5041 gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac
     5101 tgggcggcgg ccaaagcggt cggacagtgc t
//
