LOCUS       sequence-356197-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with four 
            Asp 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-4D
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..150
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(151..219,220..1011)
                     /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(369..388)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1182..1770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1671..1690
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1924..1941
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1956..2095
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2021..2040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2181..2203
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2197..2388)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3850..3869
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3953..3972
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3953..3971
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4003..4025
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4045..4062
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4072..4096
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4099..4119
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4532..4556)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4609..4627)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4623..4670
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5036..5041,5042..5058,5059..5064))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5091..5109)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
       61 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      121 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      181 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      241 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      301 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      361 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
      421 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      481 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
      541 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      601 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
      661 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
      721 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
      781 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
      841 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
      901 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
      961 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1021 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1081 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1141 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1201 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1261 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1321 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1381 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1441 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1501 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1561 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1621 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1681 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1741 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1801 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1861 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1921 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     1981 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     2041 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     2101 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2161 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2221 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     2281 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     2341 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     2401 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     2461 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     2521 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     2581 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     2641 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     2701 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     2761 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     2821 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     2881 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     2941 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     3001 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     3061 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
     3121 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt
     3181 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
     3241 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     3301 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     3361 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     3421 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     3481 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     3541 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     3601 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg atcgacgctc
     3661 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     3721 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     3781 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     3841 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     3901 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     3961 ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa
     4021 ggagatatac atatgtcgta ctaccatcac catcaccatc acctcgaatc aacaagtttg
     4081 tacaaaaaag caggcttcga gaacctgtat tttcagggta gcatggcaag cgcaagcagc
     4141 agccagcgtg gtcgtagcgg tagcggtaat tttggtggtg gtcgcggtgg tggttttggt
     4201 ggcaatggca attttggccg tggtggtaat tttagcggtc gtggtggctt tggcggtagc
     4261 cgtggcggtg gcggttatgg cggtagtggt ggtggctata atggttttgg caatagcggt
     4321 tcaaactttg gaggtggtgg cagttataac ggctttggta attacaataa tcagagcagc
     4381 aactttggtc cgatgaaagg tggcaatttc ggtggtcgtt caagcggtcc gtatggtggc
     4441 ggaggtcagt attttgccaa accgcgtaat caaggtggct atggtggttc tagcagcagc
     4501 agctcatatg gtagtggtcg tcgtttttaa gacccagctt tcttgtacaa agtggttgat
     4561 tcgaggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     4621 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     4681 gaactatatc cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg
     4741 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
     4801 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
     4861 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
     4921 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
     4981 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
     5041 agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat
     5101 acgcctattt ttataggtta atgtcatgat a
//
