LOCUS       sequence-356143-        5323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with eight 
            Asp residues added (LCD: low-complexity domain, aa 186-320).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5323)
  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 5323)
  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+8D
FEATURES             Location/Qualifiers
     source          1..5323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(161..229,230..1021)
                     /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(379..398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1192..1780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1681..1700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1934..1951
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1966..2105
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2031..2050)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2191..2213
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2207..2398)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3860..3879
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3963..3982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3963..3981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4013..4035
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4055..4072
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4073..4096
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    4274..4298
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4301..4321
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4734..4758)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4811..4829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4825..4872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5238..5243,5244..5260,5261..5266))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5293..5311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
       61 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      121 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      181 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      241 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      301 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      361 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
      421 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      481 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
      541 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
      601 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
      661 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
      721 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
      781 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
      841 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
      901 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
      961 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1021 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1321 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1801 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1861 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1921 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     1981 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     2041 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     2101 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2161 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2221 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     2281 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     2341 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     2401 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     2461 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     2521 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     2581 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     2641 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     2701 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     2761 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     2821 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     2881 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     2941 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     3001 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     3061 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     3121 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat
     3181 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     3241 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     3301 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc
     3361 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     3421 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     3481 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     3541 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     3601 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     3661 atcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     3721 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     3781 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     3841 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     3901 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     3961 attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
     4021 actttaagaa ggagatatac atatgtcgta ctaccatcac catcaccatc acctggaagt
     4081 gctgtttcag ggcccggata cctataaact ggtgattgtg ctgaacggca ccacctttac
     4141 ctataccacc gaagcggtgg atgcggcgac cgcggaaaaa gtgtttaaac agtatgcgaa
     4201 cgatgcgggc gtggatggcg aatggaccta tgatgcggcg accaaaacct ttaccgtgac
     4261 cgaactcgaa tcaacaagtt tgtacaaaaa agcaggcttc gagaacctgt attttcaggg
     4321 tagcatggca agcgcaagca gcagccagcg tgatcgtagc ggtagcggta attttggtgg
     4381 tggtcgtgat ggtggttttg gtggcaatga taattttggc cgtggtgata actttagcgg
     4441 tcgtggtgat tttggcggtt cacgtgatgg cggtggttat ggcggaagcg gtgatggtta
     4501 taatggcttt ggtaatgatg gtagcaactt tggtggcggt ggcagctata atgattttgg
     4561 taactataat aaccagagca gcaattttgg tccgatgaaa ggtggcaatt tcggtggtcg
     4621 tagcagcgat ccgtatggtg gtggcggaca gtattttgcc aaaccgcgta atcaggatgg
     4681 ttatggtggt agcagcagca gctcaagcta tgatagtggt cgtcgttttt aagacccagc
     4741 tttcttgtac aaagtggttg attcgaggct gctaacaaag cccgaaagga agctgagttg
     4801 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4861 aggggttttt tgctgaaagg aggaactata tccggatatc cacaggacgg gtgtggtcgc
     4921 catgatcgcg tagtcgatag tggctccaag tagcgaagcg agcaggactg ggcggcggcc
     4981 aaagcggtcg gacagtgctc cgagaacggg tgcgcataga aattgcatca acgcatatag
     5041 cgctagcagc acgccatagt gactggcgat gctgtcggaa tggacgatat cccgcaagag
     5101 gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag
     5161 gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa
     5221 ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattctt
     5281 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taa
//
