LOCUS       sequence-356195-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with 
            twenty-three Ser residues replaced with Gly, twelve Phe residues 
            replaced with Tyr (LCD: low-complexity domain, aa 186-320).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  AUTHORS   Bremer A, Farag M, Borcherds WM, Peran I, Martin EW, Pappu RV, 
            Mittag T
  TITLE     Deciphering how naturally occurring sequence features impact the 
            phase behaviours of disordered prion-like domains.
  JOURNAL   Nat Chem. 2022 Feb;14(2):196-207. doi: 10.1038/s41557-021-00840-w. 
            Epub 2021 Dec 20.
  PUBMED    34931046
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: 
            "10.1038/s41557-021-00840-w"; journalName: "Nat Chem"; date: 
            "2022-02"; volume: "14"; issue: "2"; pages: "196-207"
COMMENT     Sequence Label: A1-LCD+23G-23S-12F+12Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(145..213,214..1005)
                     /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(363..382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1176..1764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1665..1684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1918..1935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1950..2089
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2015..2034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2175..2197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2191..2382)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3844..3863
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3947..3966
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3947..3965
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3997..4019
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4039..4056
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4066..4090
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4093..4113
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4526..4550)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4603..4621)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4617..4664
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5030..5035,5036..5052,5053..5058))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5085..5103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
       61 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      121 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      181 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      241 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      301 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      361 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
      421 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      481 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
      541 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
      601 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
      661 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
      721 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
      781 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
      841 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
      901 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
      961 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1021 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1081 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1141 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1201 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1261 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1321 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1381 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1441 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1501 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1561 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1621 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1681 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1741 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1801 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1861 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     1921 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     1981 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2041 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     2101 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2161 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     2221 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     2281 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     2341 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     2401 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     2461 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     2521 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     2581 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     2641 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     2701 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     2761 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     2821 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg
     2881 cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc
     2941 attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc
     3001 gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac
     3061 gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg
     3121 ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg
     3181 gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac
     3241 agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg
     3301 aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc
     3361 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     3421 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     3481 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     3541 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     3601 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgatcgac gctctccctt
     3661 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     3721 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     3781 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     3841 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     3901 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     3961 atagggagac cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat
     4021 atacatatgt cgtactacca tcaccatcac catcacctcg aatcaacaag tttgtacaaa
     4081 aaagcaggct tcgagaacct gtattttcag ggtagcatgg ctggtgccgg tggtggtcag
     4141 cgtggtcgtg gtggtggcgg taattatggc ggtggtcgcg gtggcggtta tggtggtaat
     4201 gataattatg gtcgcggagg taactatggt ggccgtggtg gctatggtgg tggacgcgga
     4261 ggtggtggat atggcggagg cggtgatggt tataatggtt atggcaatga tggtggcaac
     4321 tacggtggcg gtggcggata taatgattat ggtaactata ataaccaagg cggtaactac
     4381 ggtccgatga aaggtggtaa ttacggtgga agaggcggag gtggcggtgg tggtggtggc
     4441 cagtattatg caaaaccgcg taatcaaggt ggttatggtg gtggcggagg cggaggcgga
     4501 tatggtggtg gtcgtcgtta ttaagaccca gctttcttgt acaaagtggt tgattcgagg
     4561 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     4621 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     4681 tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca
     4741 agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg
     4801 ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg
     4861 atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     4921 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
     4981 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
     5041 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
     5101 atttttatag gttaatgtca tgataataat g
//
