LOCUS       sequence-334790-        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 Patchy
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(221..245)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(298..316)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      312..359
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(725..730,731..747,748..753))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(780..798)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        866..970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(971..1039,1040..1831)
                     /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(1189..1208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2002..2590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2491..2510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2744..2761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2776..2915
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2841..2860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3001..3023
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3017..3208)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4670..4689
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4773..4792
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4773..4791
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4823..4845
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4865..4882
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4892..4916
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4919..4939
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 gattatttct actttggtag caatggcggt ggcggtagca atgatggtaa taacaatcag
       61 agcagcaatg gtccgatgaa aggtggcaat ttttactttt ttggtggtcg cagcagcggt
      121 ggttcaggtg gtggtggcca ggcaaaaccg cgtaatcaag gtggtggcgg aagctatttt
      181 ttctttagca gcagcagctc aggtagcggt cgtcgttaag acccagcttt cttgtacaaa
      241 gtggttgatt cgaggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      301 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      361 tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag
      421 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
      481 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
      541 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
      601 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
      661 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
      721 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg
      781 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
      841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1081 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1141 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1201 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1261 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1321 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1381 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1441 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1501 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1561 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1621 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1681 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1741 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1801 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1861 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1921 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1981 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2041 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2101 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2161 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2221 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2281 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2341 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2401 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2461 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2521 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2581 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2641 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2701 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2761 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2821 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     2881 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     2941 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3001 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3061 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3121 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3181 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3241 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     3301 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     3361 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     3421 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     3481 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     3541 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     3601 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     3661 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     3721 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     3781 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     3841 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     3901 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     3961 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     4021 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     4081 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     4141 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     4201 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4261 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4321 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4381 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     4441 tgggttgaag gctctcaagg gcatcggtcg atcgacgctc tcccttatgc gactcctgca
     4501 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     4561 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     4621 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     4681 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     4741 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca
     4801 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgtcgta
     4861 ctaccatcac catcaccatc acctcgaatc aacaagtttg tacaaaaaag caggcttcga
     4921 aaacctgtat tttcagggta gcatggcaag cgcaagcagc agccagcgtg gtcgttattt
     4981 ctattttagc ggtagcggta atggtggtgg tcgtggtggc ggtggtggta atgataatgg
     5041 tcgcggtggt aatagctttt atttcggtcg cggaggtggt ggttcacgcg gtggcggagg
     5101 cggaggttct ggtgatggta atggcggtaa t
//
