LOCUS       sequence-356410-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with four 
            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 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"
     protein_bind    complement(251..275)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(328..346)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      342..389
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(755..760,761..777,778..783))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(810..828)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        896..1000
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1001..1069,1070..1861)
                     /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(1219..1238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2032..2620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2521..2540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2774..2791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2806..2945
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2871..2890)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3031..3053
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3047..3238)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4700..4719
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4803..4822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4803..4821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4853..4875
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4895..4912
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4922..4946
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4949..4969
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ggttcaggtg atggttataa tggctttggt aatgatggta gcaactttgg aggtggtggt
       61 agctataatg atttcggcaa ttataacaac cagagcagca attttggtcc gatgaaaggt
      121 ggcaatttcg gtggtcgtag cagcgatccg tatggtggtg gcggtcagta ttttgccaaa
      181 ccgcgtaatc aaggtggcta tggtggttct agcagcagca gctcatatga tagtggtcgt
      241 cgtttttaag acccagcttt cttgtacaaa gtggttgatt cgaggctgct aacaaagccc
      301 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      361 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac
      421 aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc
      481 aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat
      541 tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg
      601 acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
      661 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
      721 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
      781 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      841 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      901 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      961 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1021 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1081 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1141 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1201 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     1261 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1321 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1381 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1441 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1501 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     1561 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1621 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1681 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1741 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1801 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1861 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1921 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1981 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2041 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2101 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2161 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2221 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2281 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2341 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2401 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2461 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2521 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2581 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2641 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2701 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2761 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2821 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     2881 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     2941 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3001 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3061 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3121 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3181 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3241 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3301 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     3361 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     3421 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     3481 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     3541 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     3601 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     3661 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     3721 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     3781 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     3841 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     3901 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     3961 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat
     4021 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     4081 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     4141 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc
     4201 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     4261 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     4321 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     4381 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     4441 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4501 atcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     4561 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     4621 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     4681 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     4741 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     4801 attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
     4861 actttaagaa ggagatatac atatgtcgta ctaccatcac catcaccatc acctcgaatc
     4921 aacaagtttg tacaaaaaag caggcttcga gaacctgtat tttcagggta gcatggcaag
     4981 cgcaagcagc agccagcgtg atcgtagcgg tagcggtaat tttggtggtg gtcgcggtgg
     5041 tggttttggt ggcaatgata attttggccg tggtggtaat tttagcggtc gtggcgattt
     5101 tggcggtagc cgtggcggtg gcggttatgg c
//
