LOCUS       sequence-356189-        5164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with twelve 
            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 5164)
  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 5164)
  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+12D
FEATURES             Location/Qualifiers
     source          1..5164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(164..232,233..1024)
                     /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(382..401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1195..1783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1684..1703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1937..1954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1969..2108
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2034..2053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2194..2216
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2210..2401)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3863..3882
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3966..3985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3966..3984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4016..4038
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4058..4075
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4085..4109
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4130..4150
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4578..4602)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4655..4673)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4669..4716
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5082..5087,5088..5104,5105..5110))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5137..5155)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
       61 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
      421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      541 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
      661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
      721 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
      781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
      841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
      901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1021 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1081 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1141 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1321 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     1381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1861 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     1921 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     1981 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     2041 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     2101 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2161 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2221 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     2281 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     2341 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     2401 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     2461 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     2521 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     2581 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     2641 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     2701 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     2761 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     2821 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     2881 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     2941 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     3001 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     3061 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     3121 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     3181 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     3241 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     3301 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     3361 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     3421 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     3481 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     3541 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     3601 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     3661 tcgatcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     3721 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     3781 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     3841 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     3901 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     3961 gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt
     4021 ttaactttaa gaaggagata tacatatgtc gtactaccat caccatcacc atcacctcga
     4081 atcaacaagt ttgtacaaaa aagcaggctc cgcggccgcc cccttcaccg aaaacctgta
     4141 ttttcagggt agcatggcaa gcgcagatag cagccagcgt gatcgtgatg atagcggtaa
     4201 ttttggtgat ggtcgtggtg gtggttttgg tggtaatgat aattttggcc gtggtggcaa
     4261 ttttagcgat cgcggtggct ttggcggtag ccgtggtgat ggcggttatg gcggtgatgg
     4321 tgatggatat aatggttttg gcaatgatgg tagcaacttt ggtggcggtg gtagctataa
     4381 cgattttggc aactataata accagagcag caatttcgat ccgatgaaag gtggtaactt
     4441 tggcgatcgt agcagcggtc cgtatgatgg tggtggccag tattttgcca aaccgcgtaa
     4501 tcaaggtggc tatggtggtt caagcagcag cagctcatat ggtagcgatc gtcgttttta
     4561 aaagggtggg cgcgccgacc cagctttctt gtacaaagtg gttgattcga ggctgctaac
     4621 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     4681 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     4741 tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga
     4801 agcgagcagg actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca
     4861 tagaaattgc atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc
     4921 ggaatggacg atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac
     4981 agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg
     5041 tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat
     5101 aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     5161 aggt
//
