LOCUS       sequence-356187-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with 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-12F+12Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(155..223,224..1015)
                     /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(373..392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1186..1774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1675..1694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1928..1945
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1960..2099
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2025..2044)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2185..2207
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2201..2392)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3854..3873
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3957..3976
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3957..3975
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4007..4029
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4049..4066
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4076..4100
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4103..4123
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4536..4560)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4613..4631)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4627..4674
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5040..5045,5046..5062,5063..5068))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5095..5113)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
       61 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      121 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      181 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
      241 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
      301 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
      361 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
      421 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
      481 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
      541 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
      601 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
      661 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
      721 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
      781 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
      841 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
      901 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
      961 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1021 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1081 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1141 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1201 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1261 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1321 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1381 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1441 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1501 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1561 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1621 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1681 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1741 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1801 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1861 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     1921 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     1981 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     2041 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     2101 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2161 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2221 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     2281 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     2341 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     2401 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     2461 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     2521 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     2581 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     2641 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     2701 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     2761 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     2821 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg
     2881 agatgcgccg cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt
     2941 tggtttgcgc attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga
     3001 atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc
     3061 gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc
     3121 gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga
     3181 agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac
     3241 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
     3301 cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc
     3361 gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     3421 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     3481 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     3541 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     3601 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgatcgac
     3661 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     3721 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     3781 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     3841 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     3901 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     3961 acgactcact atagggagac cacaacggtt tccctctaga aataattttg tttaacttta
     4021 agaaggagat atacatatgt cgtactacca tcaccatcac catcacctcg aatcaacaag
     4081 tttgtacaaa aaagcaggct tcgagaacct gtattttcag ggtagcatgg caagcgcaag
     4141 cagcagccag cgtggtcgta gcggtagcgg taattatggt ggtggtcgcg gtggtggtta
     4201 tggtggcaat gataattatg gccgtggtgg taattatagc ggtcgtggtg gctatggcgg
     4261 tagccgtggc ggtggcggat atggcggttc aggtgatggt tataatggct atggtaatga
     4321 tggtagcaac tatggtggcg gaggtagcta taatgattat ggcaattata acaaccagag
     4381 cagcaattac ggtccgatga aaggtggtaa ctacggtggt cgttcaagcg gtggtagcgg
     4441 aggtggcggt cagtattatg caaaaccgcg taatcaaggc ggttacggtg gttctagcag
     4501 cagcagctca tatggttcag gtcgtcgtta ttaagaccca gctttcttgt acaaagtggt
     4561 tgattcgagg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     4621 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     4681 ggaggaacta tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat
     4741 agtggctcca agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc
     4801 tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata
     4861 gtgactggcg atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat
     4921 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     4981 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     5041 ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
     5101 tgatacgcct atttttatag gttaatgtca t
//
