LOCUS       sequence-334851-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human hnRNPA1-A Low complexity domain 
            including amino acids 186-320.
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 WT
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(276..300)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(353..371)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      367..414
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(780..785,786..802,803..808))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(835..853)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        921..1025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1026..1094,1095..1886)
                     /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(1244..1263)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2057..2645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2546..2565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2799..2816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2831..2970
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2896..2915)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3056..3078
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3072..3263)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4725..4744
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4828..4847
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4828..4846
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4878..4900
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4920..4937
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4947..4971
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4974..4994
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ctctagaggc ggaggcggct atggcggaag cggcgacggc tataacggct tcggcaacga
       61 tggctccaac ttcggaggcg ggggaagcta caacgatttc ggcaactaca acaaccagag
      121 cagcaacttc ggccccatga agggcggaaa ctttggcggc agaagcagcg gcggatctgg
      181 cggaggggga cagtacttcg ccaagcccag aaaccagggc ggctacgggg gatctagcag
      241 cagctctagc tacggctccg gcagacggtt ctaagaccca gctttcttgt acaaagtggt
      301 tgattcgagg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
      361 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
      421 ggaggaacta tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat
      481 agtggctcca agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc
      541 tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata
      601 gtgactggcg atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat
      661 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
      721 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
      781 ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
      841 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
      901 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1021 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1081 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1141 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1201 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1261 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1321 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1381 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1441 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1501 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1561 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1621 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1681 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1741 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1801 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1861 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1921 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1981 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2041 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2101 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2161 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     2221 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2281 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2341 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2401 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2461 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2521 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2581 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2641 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2701 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2761 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2821 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2881 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     2941 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3001 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3061 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     3121 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     3181 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     3241 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     3301 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     3361 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     3421 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     3481 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     3541 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     3601 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     3661 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     3721 cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg
     3781 gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag
     3841 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
     3901 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     3961 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     4021 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
     4081 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
     4141 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
     4201 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     4261 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4321 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4381 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4441 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     4501 tgaaggctct caagggcatc ggtcgatcga cgctctccct tatgcgactc ctgcattagg
     4561 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     4621 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     4681 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     4741 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     4801 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt
     4861 ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg tcgtactacc
     4921 atcaccatca ccatcacctc gaatcaacaa gtttgtacaa aaaagcaggc ttcgaaaacc
     4981 tgtattttca gggcagcatg gccagcgcca gcagcagcca gagaggcaga agcggcagcg
     5041 gcaatttcgg cggaggaaga ggcggcggat tcggcggcaa cgacaacttt ggcagaggcg
     5101 gcaacttcag cggcaggggc ggatttggag g
//
