LOCUS       sequence-356307-        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 (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
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(308..332)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(385..403)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      399..446
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(812..817,818..834,835..840))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(867..885)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        953..1057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1058..1126,1127..1918)
                     /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(1276..1295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2089..2677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2578..2597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2831..2848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2863..3002
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2928..2947)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3088..3110
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3104..3295)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4757..4776
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4860..4879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4860..4878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4910..4932
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4952..4969
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4979..5003
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5006..5026
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ggtaactttg gtggccgtgg tggctttggt ggtggacgcg gaggtggtgg ttatggtggc
       61 ggaggtgatg gctataatgg ttttggcaat gatggtggca attttggagg cggtggtgga
      121 tataacgatt tcggcaatta taacaatcaa ggtggtaatt tcggtccgat gaaaggtggc
      181 aacttcggtg gaagaggtgg cggtggcggt ggtggcggag gccagtattt tgccaaaccg
      241 cgtaatcaag gcggatatgg cggaggcggt ggcggaggtg gttatggcgg tggtcgtcgt
      301 ttttaagacc cagctttctt gtacaaagtg gttgattcga ggctgctaac aaagcccgaa
      361 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      421 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg
      481 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg
      541 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc
      601 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg
      661 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
      721 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
      781 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
      841 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
      901 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
      961 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1021 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1081 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1141 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1201 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1261 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     1321 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1381 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1441 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1501 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1561 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1621 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1681 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1741 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1801 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2221 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2761 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2821 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2881 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2941 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3001 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3061 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3121 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3181 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3241 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3301 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3361 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3421 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3481 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3541 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3601 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3661 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3721 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     3781 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     3841 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     3901 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     3961 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     4021 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     4081 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     4141 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     4201 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     4261 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     4321 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     4381 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     4441 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     4501 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgatc
     4561 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4621 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     4681 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     4741 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     4801 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     4861 aatacgactc actataggga gaccacaacg gtttccctct agaaataatt ttgtttaact
     4921 ttaagaagga gatatacata tgtcgtacta ccatcaccat caccatcacc tcgaatcaac
     4981 aagtttgtac aaaaaagcag gcttcgagaa cctgtatttt cagggtagca tggctggtgc
     5041 cggtggtggt cagcgtggtc gtggtggtgg cggtaatttt ggcggtggtc gcggtggcgg
     5101 ttttggtggt aatgataatt ttggtcgcgg a
//
