LOCUS       sequence-356154-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with seven 
            Tyr residues 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+7F-7Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(177..245,246..1037)
                     /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(395..414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1208..1796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1697..1716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1950..1967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1982..2121
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2047..2066)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2207..2229
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2223..2414)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3876..3895
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3979..3998
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3979..3997
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4029..4051
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4071..4088
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4098..4122
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4125..4145
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4558..4582)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4635..4653)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4649..4696
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5062..5067,5068..5084,5085..5090))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(join(5117..5131,1..4))
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
       61 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
      421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      721 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1321 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     1381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1861 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1921 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     1981 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     2041 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     2101 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2161 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2221 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     2281 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     2341 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     2401 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     2461 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     2521 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     2581 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     2641 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     2701 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     2761 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     2821 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     2881 ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
     2941 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
     3001 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg
     3061 aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg
     3121 cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga
     3181 cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct
     3241 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
     3301 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg
     3361 ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc
     3421 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     3481 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     3541 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     3601 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     3661 tcaagggcat cggtcgatcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     3721 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     3781 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     3841 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     3901 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
     3961 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
     4021 gaaataattt tgtttaactt taagaaggag atatacatat gtcgtactac catcaccatc
     4081 accatcacct cgaatcaaca agtttgtaca aaaaagcagg cttcgagaac ctgtattttc
     4141 agggtagcat ggcaagcgca agcagcagcc agcgtggtcg tagcggtagc ggtaattttg
     4201 gtggtggtcg cggtggtggt tttggtggca atgataattt tggccgtggt ggtaatttta
     4261 gcggtcgtgg tggctttggc ggtagccgtg gcggtggcgg atttggcggt tcaggtgatg
     4321 gttttaatgg ctttggtaat gatggtagca actttggagg tggtggtagc tttaatgatt
     4381 ttggcaattt taacaaccag agcagcaatt tcggtccgat gaaaggtggt aacttcggtg
     4441 gtcgttcaag cggtggtagc ggaggtggcg gtcagttttt tgccaaaccg cgtaatcaag
     4501 gcggttttgg aggtagcagc agcagctcaa gctttggttc aggtcgtcgt ttttaagacc
     4561 cagctttctt gtacaaagtg gttgattcga ggctgctaac aaagcccgaa aggaagctga
     4621 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     4681 cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg acgggtgtgg
     4741 tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc
     4801 ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat
     4861 atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg atatcccgca
     4921 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
     4981 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
     5041 ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat
     5101 tcttgaagac gaaagggcct cgtgatacgc c
//
