LOCUS       sequence-356407-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with twenty 
            Gly residues replaced with Ser, 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-20G+20S+7F-7Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     221..240
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        221..239
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             271..293
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             313..330
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    340..364
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             367..387
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(800..824)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(877..895)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      891..938
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1304..1309,1310..1326,1327..1332))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1359..1377)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1445..1549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1550..1618,1619..2410)
                     /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(1768..1787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2581..3169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3070..3089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3323..3340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3355..3494
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3420..3439)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3580..3602
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3596..3787)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
       61 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
      121 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
      181 gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg
      241 agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg agatatacat
      301 atgtcgtact accatcacca tcaccatcac ctcgaatcaa caagtttgta caaaaaagca
      361 ggcttcgaga acctgtattt tcagggtagc atggcaagcg caagcagcag ccagcgtagc
      421 cgtagcggta gtggtaattt tagcggtagc cgttcaggta gctttagcgg caatgataat
      481 tttggtcgca gcggtaattt ttcaggtcgt agcggttttg gtggttcacg tagtggtggt
      541 ggttttagtg gtagcggtga tagctttaat agctttggta atgatggcag caatttttct
      601 ggtagcggca gctttaatga tttcggcaat tttaacaatc agagcagcaa ctttggtccg
      661 atgaaaagtg gcaattttgg tggccgtagc agcggtagca gtggtggtag tggtcagttt
      721 tttgccaaac cgcgtaatca gggttcattt agcggttcta gcagcagcag ctcatttggt
      781 agcagccgtc gtttttaaga cccagctttc ttgtacaaag tggttgattc gaggctgcta
      841 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
      901 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
      961 gatatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc
     1021 gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg
     1081 catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg
     1141 tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct
     1201 acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac
     1261 ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg
     1321 ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt
     1381 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1441 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1501 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1561 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1621 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1681 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1741 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     1801 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1861 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1921 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1981 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2041 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     2101 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2161 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2221 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2281 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2341 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2401 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2701 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3241 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3301 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3361 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     3421 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3481 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3541 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3601 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3661 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3721 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3781 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     3841 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     3901 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     3961 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     4021 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     4081 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     4141 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     4201 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     4261 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
     4321 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     4381 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     4441 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     4501 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     4561 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     4621 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     4681 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     4741 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4801 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4861 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4921 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4981 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5041 catcggtcga tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     5101 ggttgaggcc gttgagcacc gccgccgcaa g
//
