LOCUS       sequence-356414-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with ten Gly
            residues replaced with Ser (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-10G+10S
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(40..87)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     83..101
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    154..178
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(591..611)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(614..638)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(648..665)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             685..707
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(738..757)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(739..757)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(841..860)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             2322..2513
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2507..2529)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2615..2754
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2670..2689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2769..2786)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2940..3528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3020..3039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3699..4490,4491..4559))
                     /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     4322..4341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4560..4664)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4732..4750
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4777..4782,4783..4799,4800..4805)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 cccgtcctgt ggatatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg
       61 tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc
      121 ttcctttcgg gctttgttag cagcctcgaa tcaaccactt tgtacaagaa agctgggtct
      181 taaaaacgac gaccagaacc atagcttgag ctgctgctgc taccgctata actaccctga
      241 ttacgcggtt tggcaaaata ctgacctgaa ccacctgagc taccgctgct acgacctcca
      301 aaattgccgc ttttcatcgg accgaaattg ctgctctggt tgttataatt gccaaaatca
      361 ttataggagc cgctaccacc aaagttgcta ccatcattac caaagccatt atagctatca
      421 ccactaccac cataaccgcc accaccacgg ctaccgccaa agccaccacg accgctaaaa
      481 ttacctgaac ggccaaaatt atcattgcca ccaaaaccac cgctacgacc accaccaaaa
      541 ttaccactac cgctacggct acgctggctg ctgcttgcgc ttgccatgct accctgaaaa
      601 tacaggttct cgaagcctgc ttttttgtac aaacttgttg attcgaggtg atggtgatgg
      661 tgatggtagt acgacatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag
      721 ggaaaccgtt gtggtctccc tatagtgagt cgtattaatt tcgcgggatc gagatctcga
      781 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
      841 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
      901 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
      961 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
     1021 gcttcctaat gcaggagtcg cataagggag agcgtcgatc gaccgatgcc cttgagagcc
     1081 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     1141 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     1201 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     1261 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     1321 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     1381 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     1441 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     1501 caaggatcgc tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg
     1561 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
     1621 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     1681 tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc
     1741 aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt
     1801 ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg
     1861 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg
     1921 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt
     1981 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa
     2041 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc
     2101 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt
     2161 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc
     2221 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca
     2281 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa
     2341 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
     2401 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
     2461 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2521 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2581 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     2641 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2701 ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     2761 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2821 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3121 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3361 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3601 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3661 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3721 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3781 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3841 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3901 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3961 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     4021 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4081 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4141 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4201 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4261 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     4321 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4381 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4441 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4501 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4561 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4621 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4681 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4741 atcacgaggc cctttcgtct tcaagaattc tcatgtttga cagcttatca tcgataagct
     4801 ttaatgcggt agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc
     4861 taacaatgcg ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt
     4921 ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc cattccgaca gcatcgccag
     4981 tcactatggc gtgctgctag cgctatatgc gttgatgcaa tttctatgcg cacccgttct
     5041 cggagcactg tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc tacttggagc
     5101 cactatcgac tacgcgatca tggcgaccac a
//
