LOCUS       sequence-356201-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with eight 
            Phe residues removed, four Tyr 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-8F+4Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(168..236,237..1028)
                     /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(386..405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1199..1787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1688..1707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1941..1958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1973..2112
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2038..2057)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2198..2220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2214..2405)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3867..3886
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3970..3989
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3970..3988
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4020..4042
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4089..4113
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4116..4136
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4549..4573)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4626..4644)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4640..4687
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5053..5058,5059..5075,5076..5081))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5108..5126)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
       61 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      121 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      181 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      241 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      301 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      361 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
      421 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      481 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      541 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      601 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      661 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
      721 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
      781 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      841 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      901 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      961 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1021 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1081 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1141 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1201 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1261 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1321 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1381 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1441 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1501 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1561 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1621 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1681 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1741 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1801 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1861 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1921 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     1981 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     2041 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2101 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2161 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2221 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     2281 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     2341 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     2401 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     2461 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     2521 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     2581 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     2641 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     2701 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     2761 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     2821 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac
     2881 ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg
     2941 ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
     3001 cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
     3061 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa
     3121 tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg
     3181 gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat
     3241 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
     3301 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
     3361 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     3421 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     3481 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     3541 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     3601 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     3661 tcggtcgatc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     3721 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     3781 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     3841 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     3901 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     3961 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct agaaataatt
     4021 ttgtttaact ttaagaagga gatatacata tgtcgtacta ccatcaccat caccatcacc
     4081 tcgaatcaac aagtttgtac aaaaaagcag gcttcgagaa cctgtatttt cagggtagca
     4141 tggcaagcgc aagcagcagc cagcgtggtc gtagcggtag cggtaatttt ggtggtggtc
     4201 gcggtggtgg ttatggtggt aatgataatg gtggccgtgg tggcaattat agcggtcgtg
     4261 gtggttttgg cggttcacgc ggtggcggtg gctatggcgg tagtggtgat ggttataatg
     4321 gcggtggtaa cgatggtagc aattatggtg gcggaggtag ctataatgat agcggtaact
     4381 ataataacca gagcagcaat tttggtccga tgaaaggtgg caactatggt ggtcgttcaa
     4441 gcggtggttc aggtggtggt ggccagtatg gtgcaaaacc gcgtaatcaa ggcggttacg
     4501 gtggtagcag cagcagctca agctatggta gtggtcgtcg tttttaagac ccagctttct
     4561 tgtacaaagt ggttgattcg aggctgctaa caaagcccga aaggaagctg agttggctgc
     4621 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     4681 ttttttgctg aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga
     4741 tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc
     4801 ggtcggacag tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta
     4861 gcagcacgcc atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg
     4921 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     4981 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     5041 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga
     5101 cgaaagggcc tcgtgatacg cctattttta t
//
