LOCUS       sequence-356315-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with 
            fourteen Gln residues removed, four Asn removed, replaced with 
            eighteen Gly (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-14N-4Q+18G
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(285..309)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(362..380)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      376..423
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(789..794,795..811,812..817))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(844..862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        930..1034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1035..1103,1104..1895)
                     /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(1253..1272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2066..2654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2555..2574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2808..2825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2840..2979
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2905..2924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3065..3087
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3081..3272)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4734..4753
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4837..4856
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4837..4855
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4887..4909
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4929..4946
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4956..4980
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4983..5003
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ttttggcggt tcacgtggtg gtggcggata tggtggtagt ggtgatggct atggtggctt
       61 cggtggtgac ggtagcggct ttggaggtgg tggttcatat ggcgattttg gaggttatgg
      121 tggcggtagc tcaggttttg gtccgatgaa aggcggaggc tttggtggtc gttcatcagg
      181 tggttcaggt ggcggaggcg gttattttgc caaacctcgc ggaggtggcg gttacggtgg
      241 cagcagcagc tcaagcagct atggtagcgg tcgtcgtttt taagacccag ctttcttgta
      301 caaagtggtt gattcgaggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      361 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      421 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
      481 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
      541 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
      601 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
      661 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
      721 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
      781 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
      841 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
      901 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      961 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1021 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1081 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1141 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1201 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1261 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1321 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1381 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1441 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1501 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1561 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     1621 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1681 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1741 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1801 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1861 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1921 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1981 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2041 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2101 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2161 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2221 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2281 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2341 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2401 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2461 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2521 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2581 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2641 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2701 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2761 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2821 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2881 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2941 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3001 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3061 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3121 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3181 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3241 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3301 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3361 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3421 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     3481 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3541 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3601 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3661 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3721 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     3781 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     3841 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     3901 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     3961 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     4021 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     4081 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     4141 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     4201 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     4261 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4321 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4381 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4441 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4501 tgactgggtt gaaggctctc aagggcatcg gtcgatcgac gctctccctt atgcgactcc
     4561 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     4621 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     4681 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     4741 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     4801 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact atagggagac
     4861 cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat atacatatgt
     4921 cgtactacca tcaccatcac catcacctcg aatcaacaag tttgtacaaa aaagcaggct
     4981 tcgagaacct gtattttcag ggtagcatgg caagcgcaag cagcagcggt cgtggtcgta
     5041 gcggtagcgg tggttttggt ggtggtcgcg gtggtggctt tggcggaggt gatggttttg
     5101 gccgtggcgg tggcttttca ggtcgtggcg g
//
