LOCUS       sequence-356258-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with two Arg
            residues removed, two Lys removed, three Asp added (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-2R-2K+3D
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             21..38
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    48..72
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             75..95
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(508..532)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(585..603)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      599..646
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1012..1017,1018..1034,1035..1040))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1067..1085)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1153..1257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1258..1326,1327..2118)
                     /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(1476..1495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2289..2877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2778..2797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3031..3048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3063..3202
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3128..3147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3288..3310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3304..3495)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4957..4976
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5060..5079
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5060..5078
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             join(5110..5131,1)
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atatacatat gtcgtactac catcaccatc accatcacct cgaatcaaca agtttgtaca
       61 aaaaagcagg cttcgagaac ctgtattttc agggtagcat ggcaagcgca agcagcagcc
      121 aggatggtcg tagcggtagc ggtaattttg gtggtggtcg cggtggtggt tttggtggca
      181 atgataattt tggccgtggt ggtaatttta gcggtcgtgg tggctttggc ggtagccgtg
      241 gcggtggcgg ttatggcggt agtggtgatg gttataatgg ctttggtaat gatggtagca
      301 actttggagg tggtggtagc tataatgatt tcggcaatta taacaaccag agcagcaatt
      361 ttggtccgat ggatggtggt aacttcggtg gtcgtagtag cggtccgtat ggtggcggag
      421 gtcagtattt tgcagatccg cgtaatcaag gtggctatgg tggttctagc agcagcagct
      481 catatggttc aggtggtcgt ttttaagacc cagctttctt gtacaaagtg gttgattcga
      541 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
      601 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
      661 tatatccgga tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc
      721 caagtagcga agcgagcagg actgggcggc ggccaaagcg gtcggacagt gctccgagaa
      781 cgggtgcgca tagaaattgc atcaacgcat atagcgctag cagcacgcca tagtgactgg
      841 cgatgctgtc ggaatggacg atatcccgca agaggcccgg cagtaccggc ataaccaagc
      901 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
      961 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     1021 tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc
     1081 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1141 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1201 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1261 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1321 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1381 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1441 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1501 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1561 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1621 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1681 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1741 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1801 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1861 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1921 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1981 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2041 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2101 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2161 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2221 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2281 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2341 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2401 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     2461 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2521 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2581 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2641 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2701 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2761 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2821 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2881 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2941 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3001 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3061 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     3121 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     3181 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3241 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3301 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     3361 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     3421 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     3481 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     3541 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3601 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3661 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3721 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3781 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3841 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3901 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3961 tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
     4021 gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt
     4081 ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc
     4141 gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg
     4201 gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg
     4261 acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc
     4321 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
     4381 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
     4441 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     4501 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     4561 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     4621 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     4681 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     4741 ctcaagggca tcggtcgatc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     4801 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     4861 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     4921 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     4981 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     5041 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
     5101 agaaataatt ttgtttaact ttaagaagga g
//
