LOCUS       sequence-356161-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with ten Arg
            residues replaced with Lys (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-10R+10K
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(150..218,219..1010)
                     /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(368..387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1181..1769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1670..1689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1923..1940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1955..2094
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2020..2039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2180..2202
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2196..2387)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3849..3868
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3952..3971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3952..3970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4002..4024
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4044..4061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4071..4095
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4098..4118
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4531..4555)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4608..4626)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4622..4669
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5035..5040,5041..5057,5058..5063))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5090..5108)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
       61 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      241 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
      301 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
      361 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
      421 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
      481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
      541 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
      601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
      661 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
      721 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
      781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
      841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
      901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
      961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1021 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1081 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1141 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1261 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1321 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     1681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     1741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     1801 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     1861 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1921 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     1981 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     2041 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     2101 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     2161 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     2221 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     2281 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     2341 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     2401 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     2461 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     2521 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     2581 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     2641 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     2701 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     2761 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     2821 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg
     2881 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     2941 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     3001 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac
     3061 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc
     3121 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta
     3181 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc
     3241 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     3301 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     3361 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     3421 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     3481 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     3541 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     3601 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga tcgacgctct
     3661 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     3721 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     3781 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     3841 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     3901 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
     3961 tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag
     4021 gagatataca tatgtcgtac taccatcacc atcaccatca cctcgaatca acaagtttgt
     4081 acaaaaaagc aggcttcgaa aacctgtatt ttcagggcag catggccagc gccagcagca
     4141 gccagaaggg caagtctggc agcggcaatt tcggcggagg aaagggcgga ggcttcggcg
     4201 gcaacgacaa ctttggcaag ggcggcaact tcagcggcaa ggggggcttt ggaggcagca
     4261 aaggcggagg cggatatggc ggatctggcg acggctacaa cggcttcggc aacgatggct
     4321 ccaacttcgg aggcggcgga agctacaatg atttcggcaa ctacaacaac cagagcagca
     4381 acttcggccc catgaagggg ggaaactttg gcggcaagag cagcggagga tctgggggag
     4441 gcggccagta cttcgccaag cccaaaaacc agggcggcta cggcggcagc agctcctcta
     4501 gctcttacgg ctccggcaag aagttctaag acccagcttt cttgtacaaa gtggttgatt
     4561 cgaggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     4621 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     4681 aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg
     4741 ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga
     4801 gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac
     4861 tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca
     4921 agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag
     4981 atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa
     5041 gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata
     5101 cgcctatttt tataggttaa tgtcatgata a
//
