LOCUS       sequence-356411-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with two Lys
            residues removed (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-2K
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(252..276)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(329..347)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      343..390
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(756..761,762..778,779..784))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(811..829)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        897..1001
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1002..1070,1071..1862)
                     /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(1220..1239)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2033..2621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2522..2541
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2775..2792
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2807..2946
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2872..2891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3032..3054
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3048..3239)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4701..4720
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4804..4823
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4804..4822
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4854..4876
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4896..4913
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4923..4947
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4950..4970
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 cggtagtggt gatggttata atggctttgg taatgatggt agcaactttg gaggtggtgg
       61 tagctataat gatttcggca attataacaa ccagagcagc aattttggtc cgatgggtgg
      121 cggaaacttt ggtggtcgtt caagcggtcc gtatggtggt ggcggtcagt attttgcagg
      181 tccgcgtaat caaggtggct atggtggttc tagcagcagc agctcatatg gtagtggtcg
      241 tcgtttttaa gacccagctt tcttgtacaa agtggttgat tcgaggctgc taacaaagcc
      301 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
      361 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatcca
      421 caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag
      481 caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa
      541 ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg
      601 gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc
      661 cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg
      721 actgcgttag caatttaact gtgataaact accgcattaa agcttatcga tgataagctg
      781 tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
      841 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
      901 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      961 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1021 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1081 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1141 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1201 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     1261 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1321 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1381 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1441 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1501 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     1561 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1621 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1681 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1741 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1801 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1861 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1921 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1981 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2041 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2101 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2161 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2221 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2281 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2341 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2401 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2461 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2521 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2581 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2641 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2701 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2761 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2821 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     2881 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     2941 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3001 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3061 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     3121 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     3181 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     3241 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     3301 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     3361 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     3421 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     3481 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     3541 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     3601 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     3661 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg
     3721 ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc
     3781 caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga
     3841 gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc
     3901 atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg
     3961 ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa
     4021 tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt
     4081 catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga
     4141 gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc
     4201 ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     4261 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     4321 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     4381 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4441 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4501 gatcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     4561 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     4621 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     4681 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     4741 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     4801 aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt
     4861 aactttaaga aggagatata catatgtcgt actaccatca ccatcaccat cacctcgaat
     4921 caacaagttt gtacaaaaaa gcaggcttcg agaacctgta ttttcagggt agcatggcaa
     4981 gcgcaagcag cagccagcgt ggtcgtagcg gtagcggtaa ttttggtggt ggtcgcggtg
     5041 gtggttttgg tggcaatgat aattttggcc gtggtggtaa ttttagcggt cgtggtggct
     5101 ttggcggtag ccgtggcggt ggcggttatg g
//
