LOCUS       sequence-357019-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with thirty 
            Gly residues replaced with Ser, twelve Phe residues replaced with 
            Tyr (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-30G+30S-12F+12Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             167..187
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(600..624)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(677..695)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      691..738
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1104..1109,1110..1126,1127..1132))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1159..1177)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1245..1349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1350..1418,1419..2210)
                     /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(1568..1587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2381..2969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2870..2889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3123..3140
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3155..3294
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3220..3239)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3380..3402
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3396..3587)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5049..5068
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggcttcgaga acctgtattt
      181 tcagggtagc atggcaagcg caagcagcag ccagcgtagc cgtagcagca gcggtaatta
      241 tagcggtagc cgttcaggta gctatagcgg caatgataat tatggtcgca gtggcaatta
      301 tagtggtcgt agcggttatt caggtagtcg tagtggtagc ggctatagtg gttcaagcga
      361 tagctataat tcctatggta atgacagcag caattattct ggttccagca gctataatga
      421 ttatggcaac tataataacc agagcagcaa ctatggtccg atgaaaagtg gtaattattc
      481 cggtcgtagc agctcaagca gtggtagcag cggtcagtat tatgcaaaac cgcgtaatca
      541 gggttcatat agcggttcta gcagcagtag cagctattca agcagccgtc gttattaaga
      601 cccagctttc ttgtacaaag tggttgattc gaggctgcta acaaagcccg aaaggaagct
      661 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      721 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca ggacgggtgt
      781 ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca ggactgggcg
      841 gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt gcatcaacgc
      901 atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga cgatatcccg
      961 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     1021 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     1081 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     1141 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1201 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1261 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1321 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1381 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1441 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1501 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1561 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     1621 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1681 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1741 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1801 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1861 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     1921 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1981 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2041 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2101 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2161 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2221 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2281 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2341 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2401 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2461 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2521 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2581 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2641 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2701 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2761 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2821 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2881 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2941 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3001 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3061 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3121 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3181 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     3241 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3301 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3361 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3421 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3481 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3541 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     3601 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     3661 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     3721 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     3781 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     3841 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     3901 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     3961 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4021 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg
     4081 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     4141 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     4201 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac
     4261 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc
     4321 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta
     4381 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc
     4441 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     4501 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     4561 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     4621 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4681 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4741 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4801 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga tcgacgctct
     4861 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     4921 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     4981 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     5041 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     5101 ccggccacga tgcgtccggc gtagaggatc g
//
