LOCUS       sequence-274905-        5448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express hnRNPF PLD with all Y changed to S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  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 5448)
  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: hnRNPF_PLD_YtoS
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..68
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     152..171
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        152..170
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    171..195
                     /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             210..232
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             257..274
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             275..295
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             464..481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(534..552)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      548..595
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      632..1087
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(719..738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     929..950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1179..1994)
                     /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(1179..1994)
                     /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     1902..1921
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1986..2005
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2116..2704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2605..2624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2858..2875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2890..3029
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2955..2974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3115..3137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3131..3322)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4131..5213)
                     /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     5175..5194
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5214..5291)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
       61 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
      121 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
      181 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
      241 gggctctgat aaaattcatc atcatcatca tcacgaaaat ctttattttc aaggtcatat
      301 gaccggtgcg agcaacggtg cgagcagcag ccaagtgatg cagggtatgg gtgtgagcgc
      361 ggcgcaagcg accagcagcg gcctggagag ccagagcgtg agcggttgca gcggtgcggg
      421 tagcagcggt cagaacagca tgggtggcag cgactaactc gagcaccacc accaccacca
      481 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      541 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      601 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
      661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      781 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      841 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      901 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      961 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1021 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     1081 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1141 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     1201 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     1261 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     1321 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     1381 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     1441 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     1501 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     1561 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     1621 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     1681 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     1741 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     1801 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     1861 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     1921 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     1981 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     2041 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2221 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2761 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2821 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2881 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     2941 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3001 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3061 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3121 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3181 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3241 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3301 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3361 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3421 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     3481 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     3541 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     3601 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     3661 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     3721 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     3781 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     3841 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     3901 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     3961 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4021 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4081 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     4141 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4201 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     4261 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     4321 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     4381 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     4441 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     4501 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4561 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4621 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4681 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     4741 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4801 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4861 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4921 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     4981 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     5041 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     5101 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     5161 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     5221 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     5281 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     5341 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     5401 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgcc
//
