LOCUS       sequence-274899-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express hnRNPA2 with 5 serine changed 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_5E
FEATURES             Location/Qualifiers
     source          1..5751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..123
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     207..226
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        207..225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    226..250
                     /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             265..287
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             312..329
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             330..350
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             822..839
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(892..910)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      906..953
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      990..1445
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1077..1096)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1287..1308
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1537..2352)
                     /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(1537..2352)
                     /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     2260..2279
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2344..2363
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2474..3062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2963..2982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3216..3233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3248..3387
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3313..3332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3473..3495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3489..3680)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4489..5571)
                     /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     5533..5552
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5572..5649)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
       61 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      121 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      181 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
      241 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggct
      301 ctgataaaat tcatcatcat catcatcacg aaaatcttta ttttcaaggt catatgggtc
      361 gtggcggtaa cttcggtttt ggtgacagcc gtggtggtgg tggtaacttt ggtccgggtc
      421 cgggcagcaa cttccgtggt ggtagcgacg gctacggcga gggccgtggt ttcggcgatg
      481 gttataacgg ttatggtggc ggtccgggcg gtggcaactt tggtggcagc ccgggttatg
      541 gtggcggtcg tggcggttat ggcggtggcg gtccgggtta tggtaaccag ggcggtggct
      601 atggtggcgg ttacgacaac tatggcggtg gcaactacgg cgagggtaac tataacgatt
      661 tcggtaacta caaccagcaa ccggagaact atggcccgat gaagagcggt aactttggtg
      721 gcgaacgtaa catgggtggc ccgtatggtg gcggtaacta tggtccgggt ggtagcggtg
      781 gtgaaggtgg ttatggcggt cgcagccgtt actaactcga gcaccaccac caccaccact
      841 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      901 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      961 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     1021 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1081 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1141 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1201 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1261 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1321 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1381 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     1441 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1501 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
     1561 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1621 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1681 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1741 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1801 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1861 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     1921 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     1981 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2041 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2101 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2161 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2221 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2281 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     2341 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     2401 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2461 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2521 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2581 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2641 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2701 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2761 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2821 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2881 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2941 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3001 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3061 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3241 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     3301 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3361 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3421 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3481 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     3541 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     3601 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     3661 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     3721 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     3781 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     3841 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     3901 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     3961 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4021 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     4081 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     4141 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     4201 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     4261 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     4321 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     4381 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     4441 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     4501 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4561 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     4621 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     4681 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4741 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     4801 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     4861 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     4921 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     4981 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5041 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5101 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5161 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5221 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5281 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5341 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5401 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5461 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5521 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     5581 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     5641 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5701 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc a
//
