LOCUS       sequence-356293-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged A1-LCD with nine 
            Phe residues removed, six Tyr 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-9F+6Y
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        49..153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(154..222,223..1014)
                     /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(372..391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1185..1773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1674..1693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1927..1944
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1959..2098
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2024..2043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2184..2206
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2200..2391)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3853..3872
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3956..3975
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3956..3974
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4006..4028
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4048..4065
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4075..4099
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4102..4122
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(4535..4559)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4612..4630)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4626..4673
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5039..5044,5045..5061,5062..5067))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5094..5112)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
       61 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      121 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      181 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      241 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      301 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      361 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
      421 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      481 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      541 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      601 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      661 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
      721 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      781 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      841 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      901 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
      961 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1021 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1081 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1141 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1201 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1261 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1321 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1381 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1441 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1501 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1561 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1621 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1681 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1741 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1801 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1861 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1921 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     1981 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     2041 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     2101 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2161 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2221 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     2281 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     2341 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     2401 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     2461 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     2521 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     2581 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     2641 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     2701 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     2761 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     2821 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     2881 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt
     2941 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa
     3001 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     3061 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg
     3121 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa
     3181 gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc
     3241 tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc
     3301 ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg
     3361 tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     3421 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     3481 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     3541 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     3601 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgatcgacg
     3661 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     3721 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     3781 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     3841 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     3901 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
     3961 cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa
     4021 gaaggagata tacatatgtc gtactaccat caccatcacc atcacctcga atcaacaagt
     4081 ttgtacaaaa aagcaggctt cgagaacctg tattttcagg gtagcatggc aagcgcaagc
     4141 agcagccagc gtggtcgtag cggtagcggt aattttggtg gtggtcgcgg tggtggttat
     4201 ggtggtaatg ataattatgg tcgtggtggc aattatagcg gtcgtggcgg ttttggtggc
     4261 agccgtggtg gcggtggcta tggcggtagt ggtgatggtt ataatggcgg tggtaacgat
     4321 ggtagcaact atggtggcgg aggtagctat aatgatagcg gtaactataa taaccagagc
     4381 agcaattttg gtccgatgaa aggtggtaat tacggtggtc gttcaagcgg tggttcaggt
     4441 ggtggtggcc agtatggtgc aaaaccgcgt aatcaaggcg gttatggcgg ttctagcagc
     4501 agcagctcat atggtagtgg tcgtcgttat taagacccag ctttcttgta caaagtggtt
     4561 gattcgaggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     4621 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     4681 gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata
     4741 gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct
     4801 ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag
     4861 tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata
     4921 accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg
     4981 ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat
     5041 taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt
     5101 gatacgccta tttttatagg ttaatgtcat g
//
