LOCUS       sequence-356191-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with seven 
            Arg residues added, ten Glu residues 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+7R+10D
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(158..226,227..1018)
                     /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(376..395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1189..1777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1678..1697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1931..1948
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1963..2102
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2028..2047)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2188..2210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2204..2395)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3857..3876
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3960..3979
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3960..3978
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4010..4032
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4052..4069
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4079..4103
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4106..4126
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4539..4563)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4616..4634)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4630..4677
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5043..5048,5049..5065,5066..5071))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5098..5116)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
       61 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      121 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      181 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      241 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      301 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
      361 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
      421 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      481 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      541 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
      601 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
      661 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
      721 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
      781 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
      841 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
      901 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
      961 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1021 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1081 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1141 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1201 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1261 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1321 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1381 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1441 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1501 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1561 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1621 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1681 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1741 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1801 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1861 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     1921 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     1981 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2041 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     2101 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2161 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2221 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     2281 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     2341 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     2401 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     2461 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     2521 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     2581 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     2641 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     2701 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     2761 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     2821 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     2881 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     2941 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     3001 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     3061 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     3121 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     3181 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     3241 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     3301 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     3361 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     3421 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     3481 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     3541 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     3601 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgatc
     3661 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     3721 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     3781 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     3841 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     3901 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     3961 aatacgactc actataggga gaccacaacg gtttccctct agaaataatt ttgtttaact
     4021 ttaagaagga gatatacata tgtcgtacta ccatcaccat caccatcacc tcgaatcaac
     4081 aagtttgtac aaaaaagcag gcttcgagaa cctgtatttt cagggtagca tggcaagcgc
     4141 agatagcagc cagcgtgatc gtgatggtcg tggtaatttt ggtgatggcc gtggcggtgg
     4201 ttttggtggt aatgataatt ttggccgtgg tggcaatttt agcgatcgtg gtggctttgg
     4261 cggtagtcgt ggtggtggtc gttatggtgg tgatggtgat cgttataatg gttttggcaa
     4321 tgatggtcgc aactttggtg gtggcggtag ctataacgat tttggcaact ataataacca
     4381 gagcagcaac ttcgatccga tgaaaggcgg taactttcgt gatcgtagca gcggtccgta
     4441 tgatcgcggt ggtcagtatt ttgccaaacc gcgtaatcaa ggtggctatg gtggtagcag
     4501 cagcagccgt agctatggta gtgatcgtcg tttttaagac ccagctttct tgtacaaagt
     4561 ggttgattcg aggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     4621 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     4681 aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga tcgcgtagtc
     4741 gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc ggtcggacag
     4801 tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta gcagcacgcc
     4861 atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg
     4921 cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc
     4981 attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc
     5041 gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc
     5101 tcgtgatacg cctattttta taggttaatg t
//
