LOCUS       sequence-334852-        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     312..331
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     415..434
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        415..433
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             465..487
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             507..524
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    534..558
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             561..581
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(994..1018)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1071..1089)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1085..1132
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1498..1503,1504..1520,1521..1526))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1553..1571)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1639..1743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1744..1812,1813..2604)
                     /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(1962..1981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2775..3363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3264..3283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3517..3534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3549..3688
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3614..3633)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3774..3796
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3790..3981)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
       61 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgatcgacgc
      121 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
      181 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
      241 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
      301 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
      361 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
      421 gactcactat agggagacca caacggtttc cctctagaaa taattttgtt taactttaag
      481 aaggagatat acatatgtcg tactaccatc accatcacca tcacctcgaa tcaacaagtt
      541 tgtacaaaaa agcaggcttc gaaaacctgt attttcaggg cagcatggcc agcgccagca
      601 gcagccagag aggcagaagc ggctctggca atagcggcgg aggaagaggc ggcggattcg
      661 gcggcaatga caattctggc agaggcggca acagcagcgg cagaggggga tttggaggct
      721 ctagaggcgg aggcggaagt ggcggaagcg gcgacggcta taatggcagc ggcaacgacg
      781 gcagcaattc cgggggaggc ggcagctcca acgacttcgg caactccaac aaccagagca
      841 gcaacagcgg ccccatgaag ggcggcaact ttggcggcag atctagcgga ggatctggcg
      901 gagggggcca gtactctgcc aagcccagaa atcagggcgg cagcggcgga tcttccagca
      961 gctctagctc tggctccggc aggcggagct aagacccagc tttcttgtac aaagtggttg
     1021 attcgaggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1081 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1141 aggaactata tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag
     1201 tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc
     1261 cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt
     1321 gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa
     1381 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
     1441 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
     1501 aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg
     1561 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     1621 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1681 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1741 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1801 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1861 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1921 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1981 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2041 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2101 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2161 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2221 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2281 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2341 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2401 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2461 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2521 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2581 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2641 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2701 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2761 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2821 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2881 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2941 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3001 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3061 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3121 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3181 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3241 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3301 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3361 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3421 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3481 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3541 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     3601 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3661 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3721 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3781 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3841 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     3901 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3961 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     4021 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     4081 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     4141 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     4201 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     4261 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     4321 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     4381 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     4441 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     4501 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     4561 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     4621 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     4681 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     4741 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     4801 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     4861 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     4921 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     4981 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     5041 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5101 ccgaaaatga cccagagcgc tgccggcacc t
//
