LOCUS       sequence-356286-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with two Arg
            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+2R
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(246..270)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(323..341)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      337..384
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(750..755,756..772,773..778))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(805..823)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        891..995
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(996..1064,1065..1856)
                     /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(1214..1233)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2027..2615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2516..2535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2769..2786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2801..2940
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2866..2885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3026..3048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3042..3233)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4695..4714
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4798..4817
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4798..4816
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4848..4870
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4890..4907
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4917..4941
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4944..4964
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 tggtgatggt tataatggtt ttcgtaatga tggcagcaac tttggtggcg gaggtcgcta
       61 taatgatttt ggcaattata acaaccagag cagcaatttc ggtccgatga aaggtggcaa
      121 ttttggaggt cgtagcagcg gtccgtatgg tggtggcggt cagtattttg ccaaaccgcg
      181 taatcaaggt ggctatggtg gttctagcag cagcagctca tatggtagtg gtcgtcgttt
      241 ttaagaccca gctttcttgt acaaagtggt tgattcgagg ctgctaacaa agcccgaaag
      301 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
      361 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac
      421 gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac
      481 tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat
      541 caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg aatggacgat
      601 atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt
      661 gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg
      721 ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac
      781 atgagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
      841 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      901 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      961 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1021 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1081 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1141 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1201 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     1261 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1321 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1381 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1441 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1501 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     1561 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1621 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1681 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1741 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1801 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1861 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1921 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1981 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2041 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2101 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2161 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2221 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2281 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2341 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2401 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2461 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2521 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2581 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2641 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2701 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2761 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     2821 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     2881 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     2941 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3001 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3061 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     3121 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     3181 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     3241 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     3301 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     3361 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     3421 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     3481 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     3541 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     3601 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     3661 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     3721 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg
     3781 ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg
     3841 aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac
     3901 cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc
     3961 cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg
     4021 aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta
     4081 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     4141 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     4201 cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     4261 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     4321 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     4381 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     4441 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgatcga
     4501 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     4561 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     4621 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     4681 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     4741 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     4801 tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt
     4861 aagaaggaga tatacatatg tcgtactacc atcaccatca ccatcacctc gaatcaacaa
     4921 gtttgtacaa aaaagcaggc ttcgagaacc tgtattttca gggtagcatg gcaagcgcaa
     4981 gcagcagcca gcgtggtcgt agcggtagcg gtaattttgg tggtggtcgc ggtggtggtt
     5041 ttggtggcaa tgataatttt ggccgtggtg gtaattttag cggtcgtggt ggctttggcg
     5101 gtagccgtgg cggtggcggt tatggcggta g
//
