LOCUS       sequence-356145-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with 
            twenty-three Ser residues replaced with Gly, seven Tyr replaced with
            Phe (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+23G-23S+7F-7Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(160..228,229..1020)
                     /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(378..397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1191..1779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1680..1699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1933..1950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1965..2104
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2030..2049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2190..2212
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2206..2397)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3859..3878
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3962..3981
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3962..3980
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4012..4034
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4054..4071
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4081..4105
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4108..4128
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4541..4565)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4618..4636)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4632..4679
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5045..5050,5051..5067,5068..5073))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5100..5118)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
       61 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      121 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      181 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      241 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      301 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      361 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
      421 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      481 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      541 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      601 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      661 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
      721 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      781 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      841 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
      901 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
      961 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1021 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1081 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1141 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1201 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1261 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1321 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1381 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1441 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1501 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1561 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1621 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1681 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1741 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1801 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1861 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1921 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     1981 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     2041 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     2101 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2161 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2221 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     2281 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     2341 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     2401 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     2461 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     2521 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     2581 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     2641 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     2701 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     2761 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     2821 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     2881 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     2941 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     3001 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     3061 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     3121 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
     3181 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     3241 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     3301 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
     3361 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     3421 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     3481 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     3541 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     3601 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     3661 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     3721 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     3781 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     3841 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     3901 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
     3961 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
     4021 ctttaagaag gagatataca tatgtcgtac taccatcacc atcaccatca cctcgaatca
     4081 acaagtttgt acaaaaaagc aggcttcgag aacctgtatt ttcagggtag catggctggt
     4141 gccggtggtg gtcagcgtgg tcgtggtggt ggcggtaatt ttggcggtgg tcgcggtggc
     4201 ggttttggtg gtaatgataa ttttggtcgc ggaggtaact ttggtggccg tggtggcttt
     4261 ggtggtggac gcggaggtgg tggtttcggt ggcggaggtg atggctttaa tggttttggc
     4321 aatgatggtg gcaattttgg aggcggtggt ggatttaacg atttcggcaa ttttaacaat
     4381 caaggtggta atttcggtcc gatgaaaggt ggcaacttcg gtggaagagg tggcggtggc
     4441 ggtggtggcg gaggccagtt ttttgccaaa ccgcgtaatc aaggcggttt tggcggaggc
     4501 ggaggtggcg gtggttttgg tggtggccgt cgtttttaag acccagcttt cttgtacaaa
     4561 gtggttgatt cgaggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     4621 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     4681 tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag
     4741 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
     4801 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
     4861 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
     4921 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     4981 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     5041 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg
     5101 cctcgtgata cgcctatttt tataggttaa t
//
