LOCUS       sequence-356186-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with 
            twenty-three Thr reisudes replaced with Ser (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-23S+23T
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    413..552
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(478..497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     638..660
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(654..845)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2307..2326
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2410..2429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2410..2428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2460..2482
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2502..2519
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2529..2553
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2556..2576
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2989..3013)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3066..3084)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3080..3127
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3493..3498,3499..3515,3516..3521))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3548..3566)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3634..3738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3739..3807,3808..4599)
                     /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(3957..3976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4770..5131,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
      421 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
      481 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
      541 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      601 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      661 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
      721 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
      781 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
      841 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
      901 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
      961 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     1021 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     1081 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     1141 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     1201 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     1261 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac
     1321 ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg
     1381 ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
     1441 cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
     1501 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa
     1561 tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg
     1621 gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat
     1681 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
     1741 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
     1801 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     1861 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     1921 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     1981 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     2041 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     2101 tcggtcgatc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     2161 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     2221 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     2281 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     2341 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     2401 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct agaaataatt
     2461 ttgtttaact ttaagaagga gatatacata tgtcgtacta ccatcaccat caccatcacc
     2521 tcgaatcaac aagtttgtac aaaaaagcag gcttcgagaa cctgtatttt cagggtagca
     2581 tggcaaccgc aaccaccaca cagcgtggtc gtaccggcac cggtaatttt ggtggtggtc
     2641 gtggtggcgg ttttggtggc aatgataatt ttggccgtgg tggtaatttt accggtcgcg
     2701 gtggttttgg cggtacacgt ggcggtggtg gctatggtgg caccggtgat ggttataatg
     2761 gttttggtaa tgatggcacc aactttggcg gtggcggtac gtataacgat tttggcaatt
     2821 ataacaacca gaccaccaat ttcggtccga tgaaaggtgg caactttggt ggtcgcacca
     2881 ctggtggtac aggtggtggt ggccagtatt ttgccaaacc gcgtaatcaa ggtggttacg
     2941 gtggtactac cacaaccaca acctatggta cgggtcgtcg tttttaagac ccagctttct
     3001 tgtacaaagt ggttgattcg aggctgctaa caaagcccga aaggaagctg agttggctgc
     3061 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3121 ttttttgctg aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga
     3181 tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc
     3241 ggtcggacag tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta
     3301 gcagcacgcc atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg
     3361 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     3421 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     3481 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga
     3541 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3601 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3661 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3721 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3781 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3841 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3901 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3961 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     4021 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4081 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4141 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4201 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4261 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     4321 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4381 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4441 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4501 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4561 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4621 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4681 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5101 tgcacacagc ccagcttgga gcgaacgacc t
//
