LOCUS       sequence-356159-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with three 
            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-3R+3K
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     77..94
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    109..248
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(174..193)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     334..356
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(350..541)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2003..2022
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2106..2125
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2106..2124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2156..2178
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2198..2215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2225..2249
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2252..2272
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2685..2709)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2762..2780)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2776..2823
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3189..3194,3195..3211,3212..3217))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3244..3262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3330..3434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3435..3503,3504..4295)
                     /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(3653..3672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4466..5054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4955..4974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
       61 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
      121 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
      181 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
      241 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      301 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      361 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
      421 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
      481 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
      541 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
      601 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
      661 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
      721 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
      781 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
      841 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
      901 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
      961 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     1021 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     1081 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     1141 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     1201 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     1261 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     1321 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     1381 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     1441 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     1501 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     1561 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     1621 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     1681 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     1741 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     1801 tcgatcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     1861 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     1921 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     1981 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     2041 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     2101 gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt
     2161 ttaactttaa gaaggagata tacatatgtc gtactaccat caccatcacc atcacctcga
     2221 atcaacaagt ttgtacaaaa aagcaggctt cgaaaacctg tattttcagg gcagcatggc
     2281 cagcgccagc agcagccaga gaggcaagtc tggcagcggc aatttcggcg gaggaagagg
     2341 cggcggattc ggcggcaacg acaactttgg cagaggcggc aacttcagcg gcaggggcgg
     2401 atttggaggc tctaaaggcg gaggcggcta tggcggaagc ggcgacggct ataacggctt
     2461 cggcaacgat ggctccaact tcggaggcgg gggaagctac aacgatttcg gcaactacaa
     2521 caaccagagc agcaacttcg gccccatgaa gggcggaaac tttggcggca gaagcagcgg
     2581 cggatctggc ggagggggac agtacttcgc caagcccaga aaccagggcg gctacggggg
     2641 atctagcagc agctctagct acggcagcgg ccggaagttc taagacccag ctttcttgta
     2701 caaagtggtt gattcgaggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     2761 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2821 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
     2881 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
     2941 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
     3001 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
     3061 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     3121 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     3181 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     3241 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3301 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3361 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3421 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3481 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3541 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3601 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3661 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3721 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3781 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3841 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3901 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3961 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     4021 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4081 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4141 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4201 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4261 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4321 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4381 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4441 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4501 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4561 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4921 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4981 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5041 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5101 cttttgctca catgttcttt cctgcgttat c
//
