LOCUS       sequence-334793-        5116 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial Expression of human hnRNPA1-A Low complexity domain 
            including amino acids 186-320, hexapeptide including aa 259-264 
            deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5116)
  AUTHORS   Martin EW, Holehouse AS, Peran I, Farag M, Incicco JJ, Bremer A, 
            Grace CR, Soranno A, Pappu RV, Mittag T
  TITLE     Valence and patterning of aromatic residues determine the phase 
            behavior of prion-like domains.
  JOURNAL   Science. 2020 Feb 7;367(6478):694-699. doi: 10.1126/science.aaw8653.
  PUBMED    32029630
REFERENCE   2  (bases 1 to 5116)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2020-02-7- 7"; volume: "367"; issue: 
            "6478"; pages: "694-699"
COMMENT     Sequence Label: A1-LCD deltahexa
FEATURES             Location/Qualifiers
     source          1..5116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(32..56)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(109..127)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      123..170
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(536..541,542..558,559..564))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(591..609)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        677..781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(782..850,851..1642)
                     /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(1000..1019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1813..2401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2302..2321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2555..2572
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2587..2726
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2652..2671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2812..2834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2828..3019)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4481..4500
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4584..4603
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4584..4602
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4634..4656
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4676..4693
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4703..4727
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4730..4750
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 agctatggca gtggcagaag attttaatag gacccagctt tcttgtacaa agtggttgat
       61 tcgaggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      121 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      181 gaactatatc cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg
      241 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
      301 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
      361 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
      421 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
      481 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
      541 agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat
      601 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
      661 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      721 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      781 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      841 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      901 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      961 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1021 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1081 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1141 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1201 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1261 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1321 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1381 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1441 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1501 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1561 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1621 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1681 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1741 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1801 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1861 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1921 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     1981 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2041 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2101 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2161 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2221 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2281 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2341 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2401 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2461 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2521 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2581 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     2641 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2701 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2761 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2821 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2881 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2941 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3001 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     3061 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     3121 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3181 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3241 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3301 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3361 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3421 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3481 cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg
     3541 acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt
     3601 gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca
     3661 ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag
     3721 ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc
     3781 cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg
     3841 aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat
     3901 cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc
     3961 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     4021 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     4081 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     4141 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     4201 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     4261 ggctctcaag ggcatcggtc gatcgacgct ctcccttatg cgactcctgc attaggaagc
     4321 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     4381 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     4441 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     4501 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     4561 tcgagatctc gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc
     4621 ctctagaaat aattttgttt aactttaaga aggagatata catatgtcgt actaccatca
     4681 ccatcaccat cacctcgaat caacaagttt gtacaaaaaa gcaggcttcg aaaacctgta
     4741 ttttcagggc agcatggcta gtgcttcatc cagccaaaga ggtcgaagtg gttctggaaa
     4801 ctttggtggt ggtcgtggag gtggtttcgg tgggaatgac aacttcggtc gtggaggaaa
     4861 cttcagtggt cgtggtggct ttggtggcag ccgtggtggt ggtggatatg gtggcagtgg
     4921 ggatggctat aatggatttg gtaatgatgg aagcaatttt ggaggtggtg gaaattacaa
     4981 caatcagtct tcaaattttg gacccatgaa gggaggaaat tttggaggca gaagctctgg
     5041 cccctatggc ggtggaggcc aatactttgc aaaaccacga aaccaaggtg gctatggcgg
     5101 ttccagcagc agcagt
//
