LOCUS       sequence-356295-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with four 
            Arg residues removed, two Lys removed, five 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-4R-2K+5D
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     227..246
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        227..245
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             277..299
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             319..336
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    346..370
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             373..393
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(806..830)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(883..901)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      897..944
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1310..1315,1316..1332,1333..1338))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1365..1383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1451..1555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1556..1624,1625..2416)
                     /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(1774..1793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2587..3175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3076..3095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3329..3346
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3361..3500
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3426..3445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3586..3608
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3602..3793)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
       61 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
      121 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
      181 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
      241 atagggagac cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat
      301 atacatatgt cgtactacca tcaccatcac catcacctcg aatcaacaag tttgtacaaa
      361 aaagcaggct tcgagaacct gtattttcag ggtagcatgg caagcgcaag cagcagccag
      421 gatggtcgta gcggtagcgg taattttggt ggtggtgatg gtggcggttt tggtggcaat
      481 gataattttg gccgtggtgg taattttagc ggtggcggtg gttttggcgg tagtcgcgga
      541 ggtggtggct atggcggttc aggtgatggt tataatggtt ttggtaatga tggcagcaac
      601 tttggtggcg gaggtagcta taacgatttt ggcaattata acaaccagag cagcaatttt
      661 ggaccgatgg atggtggtaa ctttggaggt cgtagcagcg gtccgtatgg cggaggcggt
      721 cagtattttg cagatccgcg taatcaaggc ggatatggtg gtagcagcag cagctcaagc
      781 tatggtagcg gtgatcgttt ttaagaccca gctttcttgt acaaagtggt tgattcgagg
      841 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
      901 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
      961 tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca
     1021 agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg
     1081 ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg
     1141 atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     1201 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
     1261 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
     1321 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
     1381 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1441 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1501 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1561 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1621 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1681 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1741 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1801 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1861 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1921 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1981 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2041 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     2101 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2161 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2221 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2281 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2341 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2401 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2461 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2521 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2581 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2641 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2701 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2761 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2821 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2881 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2941 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3001 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3061 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3121 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3181 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3241 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3301 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3361 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3421 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3481 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3541 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3601 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3661 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3721 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3781 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3841 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3901 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3961 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     4021 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     4081 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     4141 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4201 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4261 gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga
     4321 tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg
     4381 ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     4441 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     4501 gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac
     4561 gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg
     4621 tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat
     4681 gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc
     4741 cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc
     4801 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4861 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4921 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     4981 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     5041 caagggcatc ggtcgatcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5101 gtagtaggtt gaggccgttg agcaccgccg c
//
