LOCUS       sequence-356275-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with twenty 
            Gly residues replaced with Ser, 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-20G+20S-12F+12Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     465..484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     718..735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    750..889
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(815..834)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     975..997
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(991..1182)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2644..2663
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2747..2766
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2747..2765
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2797..2819
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2839..2856
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2866..2890
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2893..2913
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(3326..3350)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3403..3421)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3417..3464
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3830..3835,3836..3852,3853..3858))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3885..3903)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3971..4075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4076..4144,4145..4936)
                     /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(4294..4313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5107..5131,1..564)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
       61 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      121 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
      721 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
      781 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      841 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
      901 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      961 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     1021 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     1081 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     1141 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     1201 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     1261 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     1321 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     1381 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     1441 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     1501 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     1561 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     1621 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg
     1681 cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc
     1741 attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc
     1801 gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac
     1861 gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg
     1921 ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg
     1981 gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac
     2041 agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg
     2101 aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc
     2161 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     2221 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     2281 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     2341 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     2401 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgatcgac gctctccctt
     2461 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     2521 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     2581 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     2641 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     2701 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     2761 atagggagac cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat
     2821 atacatatgt cgtactacca tcaccatcac catcacctcg aatcaacaag tttgtacaaa
     2881 aaagcaggct tcgagaacct gtattttcag ggtagcatgg caagcgcaag cagcagccag
     2941 cgtagccgta gcggtagtgg taattatagc ggtagccgtt caggtagcta tagcggcaat
     3001 gataattatg gtcgcagcgg taattattca ggtcgtagcg gttatggtgg ttcacgtagt
     3061 ggtggtggtt atagtggtag cggtgatagc tataattcct atggtaatga tggcagcaat
     3121 tattctggta gcggcagcta taatgattac ggcaattata acaatcagag cagcaactat
     3181 ggtccgatga aaagtggcaa ttatggtggc cgtagcagcg gtagcagtgg tggtagtggt
     3241 cagtattatg caaaaccgcg taatcagggt tcatatagcg gttctagcag cagcagctca
     3301 tatggtagca gccgtcgtta ttaagaccca gctttcttgt acaaagtggt tgattcgagg
     3361 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     3421 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     3481 tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca
     3541 agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg
     3601 ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg
     3661 atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     3721 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
     3781 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
     3841 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
     3901 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3961 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4021 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4081 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4141 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4201 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4261 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     4321 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4381 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4441 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4501 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4561 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     4621 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4681 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4741 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4801 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4861 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4921 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5101 atcttcttga gatccttttt ttctgcgcgt a
//
