LOCUS       sequence-356310-        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 (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
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(186..206)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(209..233)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(243..260)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             280..302
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(333..352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(334..352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(436..455)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             1917..2108
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2102..2124)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2210..2349
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2265..2284
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2364..2381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2535..3123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2615..2634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3294..4085,4086..4154))
                     /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     3917..3936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4155..4259)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4327..4345
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4372..4377,4378..4394,4395..4400)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(4766..4813)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4809..4827
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    4880..4904
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ctattatagc tatcgctact accgctataa ccgctaccac tacgactacc tgaaaaaccg
       61 ctacgaccac taaaattgcc actgcgacca aaattatcat tgccgctaaa gctacctgaa
      121 cggctaccgc taaaattacc gctgctgcta cggctacgct ggctgctgct tgcgcttgcc
      181 atgctaccct gaaaatacag gttctcgaag cctgcttttt tgtacaaact tgttgattcg
      241 aggtgatggt gatggtgatg gtagtacgac atatgtatat ctccttctta aagttaaaca
      301 aaattatttc tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg
      361 ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
      421 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
      481 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
      541 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
      601 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgatcgaccg
      661 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
      721 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
      781 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
      841 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
      901 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
      961 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     1021 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     1081 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga
     1141 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
     1201 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     1261 cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa
     1321 gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga
     1381 acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg
     1441 ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc
     1501 ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact
     1561 gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt
     1621 cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg
     1681 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt
     1741 gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac
     1801 aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc
     1861 gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga
     1921 ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc
     1981 ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc
     2041 acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa
     2101 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2161 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     2221 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     2281 tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2341 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2401 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2461 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2701 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2941 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3601 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3901 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4261 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     4321 tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattctcatg tttgacagct
     4381 tatcatcgat aagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc
     4441 accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc
     4501 tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc
     4561 cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct
     4621 atgcgcaccc gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc
     4681 ttcgctactt ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat
     4741 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
     4801 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
     4861 tgttagcagc ctcgaatcaa ccactttgta caagaaagct gggtcttaaa aacgacgact
     4921 gctgctatag ctagagctgc ttgatgaacc gctatatgaa ccctgattac gcggtttggc
     4981 aaaatactga ccgcttgaac cgctgcttga gctgctacga ccggaaaaat taccactttt
     5041 catcggacca aagttgctgc tctggttgtt ataattgccg aaatcattat agctgctgga
     5101 accagaaaaa ttgctgctgt cattaccaaa g
//
