LOCUS       sequence-274902-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express hnRNPA2 with all serine change to glutamate .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  AUTHORS   Ryan VH, Perdikari TM, Naik MT, Saueressig CF, Lins J, Dignon GL, 
            Mittal J, Hart AC, Fawzi NL
  TITLE     Tyrosine phosphorylation regulates hnRNPA2 granule protein 
            partitioning and reduces neurodegeneration.
  JOURNAL   EMBO J. 2021 Feb 1;40(3):e105001. doi: 10.15252/embj.2020105001. 
            Epub 2020 Dec 22.
  PUBMED    33349959
REFERENCE   2  (bases 1 to 5751)
  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"; journalName: "EMBO J."; 
            date: "2021-02-1"; volume: "40(3):e105001. doi"; pages: " 
            10.15252/embj.2020105001. Epub 2020 Dec 22"
COMMENT     Sequence Label: hnRNPA2_LC_12E
FEATURES             Location/Qualifiers
     source          1..5751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..112
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     196..215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        196..214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    215..239
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             254..276
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             301..318
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             319..339
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             811..828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(881..899)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      895..942
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      979..1434
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1066..1085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1276..1297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1526..2341)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1526..2341)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2249..2268
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2333..2352
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2463..3051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2952..2971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3205..3222
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3237..3376
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3302..3321)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3462..3484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3478..3669)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4478..5560)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5522..5541
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5561..5638)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
       61 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
      121 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
      181 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      241 ctctagaaat aattttgttt aactttaaga aggagatata ccatgggctc tgataaaatt
      301 catcatcatc atcatcacga aaatctttat tttcaaggtc atatgggtcg tggcggtaac
      361 tttggtttcg gtgatgagcg tggcggcggc ggtaactttg gtccgggtcc gggcgagaac
      421 tttcgtggcg gtgaggacgg ctacggtgaa ggccgtggtt tcggcgatgg ttataacggt
      481 tatggtggcg gtccgggcgg tggcaacttt ggtggcgagc cgggttatgg tggcggtcgt
      541 ggcggttatg gcggtggcgg tccgggttat ggtaaccagg gcggtggcta tggtggcggt
      601 tacgacaact atggcggtgg caactacggc gagggtaact ataacgattt cggtaactac
      661 aaccagcaac cggaaaacta tggcccgatg aaagagggta actttggtgg cgaacgtaac
      721 atgggtggcc cgtatggtgg cggtaactac ggtccgggcg gcgaaggcgg cgaaggcggt
      781 tacggcggtc gtgaacgcta ctaactcgag caccaccacc accaccactg agatccggct
      841 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
      901 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
      961 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1021 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1081 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1141 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1201 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1261 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1321 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1381 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     1441 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1501 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca
     1561 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1621 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1681 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1741 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt
     1801 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1861 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     1921 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     1981 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     2041 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     2101 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     2161 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     2221 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2281 catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca
     2341 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc
     2401 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2461 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2521 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2581 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2641 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2701 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2761 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2821 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2881 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2941 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3001 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3061 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3121 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3181 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3241 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     3301 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3361 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3481 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3541 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3601 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3661 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     3721 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3781 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3841 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     3901 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     3961 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     4021 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     4081 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     4141 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     4201 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4261 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4321 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     4381 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4441 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4561 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4621 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4681 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4741 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4801 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4861 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4921 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     4981 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     5041 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     5101 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5161 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5221 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5281 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5341 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5401 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5461 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5521 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5581 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5641 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     5701 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc g
//
