LOCUS       sequence-356435-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with nine 
            Phe residues removed, three 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-9F+3Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(191..215)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(268..286)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      282..329
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(695..700,701..717,718..723))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(750..768)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        836..940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(941..1009,1010..1801)
                     /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(1159..1178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1972..2560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2461..2480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2714..2731
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2746..2885
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2811..2830)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2971..2993
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2987..3178)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4640..4659
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4743..4762
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4743..4761
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4793..4815
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4835..4852
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4862..4886
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4889..4909
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 agctataatg atagcggcaa tggtaataat cagagcagca attttggtcc gatgaaaggt
       61 ggcaactatg gtggtcgttc aagcggtggt tcaggtggtg gtggccagta tggtgcaaaa
      121 ccgcgtaatc aaggcggtta cggtggtagc agcagcagct caagctatgg tagtggtcgt
      181 cgtagctaag acccagcttt cttgtacaaa gtggttgatt cgaggctgct aacaaagccc
      241 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      301 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac
      361 aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc
      421 aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat
      481 tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg
      541 acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
      601 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
      661 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
      721 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      781 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      841 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      901 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      961 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1021 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1081 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1141 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     1201 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1261 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1321 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1381 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1441 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     1501 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1561 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1621 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1681 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1741 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1801 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1861 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1921 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1981 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2041 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2101 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2161 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2221 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2281 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2341 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2401 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2461 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2521 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2581 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2641 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2701 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2761 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     2821 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     2881 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2941 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3001 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3061 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3121 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3181 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3241 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     3301 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     3361 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     3421 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     3481 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     3541 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     3601 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     3661 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     3721 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     3781 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     3841 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     3901 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat
     3961 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     4021 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     4081 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc
     4141 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     4201 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     4261 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     4321 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     4381 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4441 atcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     4501 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     4561 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     4621 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     4681 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     4741 attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
     4801 actttaagaa ggagatatac atatgtcgta ctaccatcac catcaccatc acctcgaatc
     4861 aacaagtttg tacaaaaaag caggcttcga gaacctgtat tttcagggta gcatggcaag
     4921 cgcaagcagc agccagcgtg gtcgtagcgg tagcggtaat tttggtggtg gtcgcggtgg
     4981 tggttatggt ggtaatgata atggtggccg tggtggcaat tatagcggtc gtggtggttt
     5041 tggcggttca cgcggtggcg gtggctatgg cggtagtggt gatggttata atggcggtgg
     5101 taacgatggt agcaattatg gtggcggagg t
//
