LOCUS       sequence-334791-        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. PHE and TYR 
            amino acids shuffled to optimize mixing..
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"
     protein_bind    complement(253..277)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(330..348)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      344..391
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(757..762,763..779,780..785))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(812..830)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        898..1002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1003..1071,1072..1863)
                     /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(1221..1240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2034..2622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2523..2542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2776..2793
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2808..2947
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2873..2892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3033..3055
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3049..3240)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4702..4721
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4805..4824
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4805..4823
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4855..4877
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4897..4914
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4924..4948
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4951..4971
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 gcggtagtgg tgatggttat aatggctttg gtaatgatgg tagcaacttt ggaggtggtg
       61 gtagctataa tgatttcggc aattataaca accagagcag caattttggt ccgatgaaag
      121 gtggcaattt cggtggtcgt agcagcggtg gttcatatgg tggtggccag tattttgcca
      181 aaccgcgtaa tcaaggtggc tatggtggta gtagctttag cagcagctat ggtagtggtc
      241 gtcgttttta agacccagct ttcttgtaca aagtggttga ttcgaggctg ctaacaaagc
      301 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
      361 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
      421 acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga
      481 gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa
      541 attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat
      601 ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
      661 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
      721 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
      781 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
      841 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
      901 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      961 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1021 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1081 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1141 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1201 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     1261 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1321 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1381 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1441 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1501 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     1561 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1621 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1681 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1741 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1801 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1861 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1921 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1981 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2041 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2101 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2161 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2221 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2281 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2341 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2401 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2461 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2521 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2581 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2641 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2701 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2761 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     2821 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     2881 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     2941 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3001 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3061 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     3121 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     3181 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     3241 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     3301 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     3361 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     3421 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     3481 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     3541 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     3601 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     3661 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     3721 gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg
     3781 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg
     3841 agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc
     3901 catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat
     3961 gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca
     4021 atgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg
     4081 tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg
     4141 agaagaatca taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag
     4201 cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     4261 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     4321 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     4381 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     4441 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     4501 cgatcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     4561 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     4621 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     4681 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     4741 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     4801 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
     4861 taactttaag aaggagatat acatatgtcg tactaccatc accatcacca tcacctcgaa
     4921 tcaacaagtt tgtacaaaaa agcaggcttc gaaaacctgt attttcaggg tagcatggca
     4981 tttgcaagca gctttcagcg tggtcgttat ggtagcggta attttggtgg tggtcgcggt
     5041 ggtggttttg gtggcaatga taattttggc cgtggtggta attttagcgg tcgtggtggc
     5101 tttggcggta gccgtggcgg tggcggttat g
//
