LOCUS       sequence-334843-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of human hnRNPA1-A Low complexity domain 
            including amino acids 186-320, aromatic amino acids. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  AUTHORS   Martin EW, Holehouse AS, Peran I, Farag M, Incicco JJ, Bremer A, 
            Grace CR, Soranno A, Pappu RV, Mittag T
  TITLE     Valence and patterning of aromatic residues determine the phase 
            behavior of prion-like domains.
  JOURNAL   Science. 2020 Feb 7;367(6478):694-699. doi: 10.1126/science.aaw8653.
  PUBMED    32029630
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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2020-02-7- 7"; volume: "367"; issue: 
            "6478"; pages: "694-699"
COMMENT     Sequence Label: A1-LCD Aro-
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    158..297
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(223..242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     383..405
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(399..590)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2052..2071
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2155..2174
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2155..2173
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2205..2227
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2247..2264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2274..2298
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2301..2321
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2734..2758)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2811..2829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2825..2872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3238..3243,3244..3260,3261..3266))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3293..3311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3379..3483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3484..3552,3553..4344)
                     /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(3702..3721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4515..5103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5004..5023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
       61 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      121 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
      181 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
      241 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
      301 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
      361 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
      421 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
      481 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
      541 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
      601 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
      661 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
      721 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
      781 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
      841 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
      901 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
      961 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     1021 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     1081 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     1141 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     1201 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     1261 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     1321 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag
     1381 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     1441 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     1501 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     1561 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     1621 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     1681 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     1741 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     1801 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgatcgacgc
     1861 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     1921 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     1981 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     2041 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     2101 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     2161 gactcactat agggagacca caacggtttc cctctagaaa taattttgtt taactttaag
     2221 aaggagatat acatatgtcg tactaccatc accatcacca tcacctcgaa tcaacaagtt
     2281 tgtacaaaaa agcaggcttc gaaaacctgt attttcaggg cagcatggcc agcgccagca
     2341 gcagccagag aggcagaagc ggctctggca atagcggcgg aggaagaggc ggcggattcg
     2401 gcggcaacga caattttggc agaggcggca acagcagcgg caggggcgga tttggaggat
     2461 ctagaggcgg aggcggctac ggcggaagcg gcgacggata caatggcttc ggcaacgatg
     2521 gcagcaacag cggagggggc ggaagcagca acgacttcgg caattacaac aaccagagca
     2581 gcaacttcgg ccccatgaag ggcggcaact ttggcggcag atctagcgga ggatctggcg
     2641 ggggaggcca gtactctgcc aagcccagaa atcagggcgg ctatgggggc agcagctcca
     2701 gctctagttc tggctccggc agacggttct aagacccagc tttcttgtac aaagtggttg
     2761 attcgaggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2821 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     2881 aggaactata tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag
     2941 tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc
     3001 cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt
     3061 gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa
     3121 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
     3181 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
     3241 aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg
     3301 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3361 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3421 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3481 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3541 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3601 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3661 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3721 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3781 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3841 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3901 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3961 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4021 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4081 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4141 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4201 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4261 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4321 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4381 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4441 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4501 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4561 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4621 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4681 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4741 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4801 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4861 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4921 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4981 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5041 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5101 aaaaacgcca gcaacgcggc ctttttacgg t
//
