LOCUS       sequence-274867-        7545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express FL hnRNPF without the prion like domain with N-terminal 
            maltose binding protein tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7545)
  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 7545)
  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: pTHMT_hnRNPF_deltaPLD
FEATURES             Location/Qualifiers
     source          1..7545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     939..958
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(978..1055)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1261..1280
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1364..1383
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1364..1382
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1383..1407
                     /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             1422..1444
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1469..1486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1499..2596
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      2453..2462
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2570..2593
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    2615..2639
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2648..2668
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3773..3790
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3843..3861)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3857..3904
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3941..4396
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4028..4047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4238..4259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4488..5303)
                     /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(4488..5303)
                     /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     5211..5230
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5295..5314
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5425..6013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5914..5933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6167..6184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6199..6338
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6264..6283)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6424..6446
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6440..6631)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7440..7545,1..977))
                     /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"
ORIGIN
        1 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
       61 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
      121 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
      181 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
      241 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
      301 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
      361 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
      421 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
      481 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
      541 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
      601 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
      661 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
      721 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
      781 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
      841 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
      901 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
      961 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     1021 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     1081 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     1141 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     1201 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     1261 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     1321 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
     1381 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
     1441 gagatatacc atgggatctg ataaaattca tcatcatcat catcacagca gcggtaccaa
     1501 aatcgaagaa ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc
     1561 tgaagtcggt aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga
     1621 taaactggaa gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt
     1681 ctgggcacac gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc
     1741 ggacaaagcg ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg
     1801 caagctgatt gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct
     1861 gctgccgaac ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc
     1921 gaaaggtaag agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat
     1981 tgctgctgac gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt
     2041 gggcgtggat aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa
     2101 caaacacatg aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga
     2161 aacagcgatg accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa
     2221 ttatggtgta acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt
     2281 gctgagcgca ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga
     2341 aaactatctg ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc
     2401 cgtagcgctg aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccaccat
     2461 ggaaaacgcc cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta
     2521 tgccgtgcgt actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct
     2581 gaaagacgcg cagactaatt cgggatctga tatcacaagt ttgtacaaaa aagctgaagg
     2641 cggtaccgag aacctgtact tccagggtca tatgatgctg ggtcctgagg gtggcgaggg
     2701 tttcgttgtc aagttgcgcg gtctgccgtg gtcttgctct gttgaagatg tccagaactt
     2761 cctgtcggat tgtactatcc acgatggcgc tgccggtgtg cacttcatct atacccgtga
     2821 gggtcgccag agcggtgagg ccttcgttga gctgggttcc gaagatgacg tcaaaatggc
     2881 gctgaaaaaa gaccgtgaat ctatgggcca ccgttatatt gaagtgttca aaagccaccg
     2941 taccgagatg gactgggttc tgaagcacag cggtccgaac agcgcagata gcgcgaatga
     3001 tggttttgtt cgtctgcgtg gcctgccttt tggttgcacc aaagaagaaa ttgtccaatt
     3061 tttctcgggt ctggaaatcg ttccgaatgg cattacgctg ccggttgatc cggaaggcaa
     3121 gattaccggt gaggcattcg tgcaatttgc tagccaagaa ctggcagaga aagcactggg
     3181 caagcacaag gaacgtattg gtcaccgcta catcgaagta ttcaagagca gccaagagga
     3241 agtgcgcagc tatagcgatc cgccgctgaa gtttatgtcc gtgcagcgtc cgggtccata
     3301 cgaccgcccg ggcaccgccc gtcgttatat cggtatcgtg aaacaggccg gcttggagcg
     3361 catgcgcccg ggtgcgtata gcacgggcta cggtggctac gaagagtact ctggcctgag
     3421 cgacggctat ggttttacga cggacctgtt cggtcgtgac ttgagctatt gcctgagcgg
     3481 tatgtacgac caccgttatg gtgattccga gtttaccgtc cagtctacca ccggtcattg
     3541 tgttcacatg cgtggtctgc catacaaggc aaccgagaat gacatctaca acttcttcag
     3601 cccgctgaat ccggtgcgtg tccacatcga aattggtccg gatggtcgtg tcacgggtga
     3661 ggctgatgtc gaatttgcga ctcatgaaga agccgtggcg gcgatgagca aagaccgtgc
     3721 gaacatgcag catcgctaca ttgagttgtt tctgaacagc acctgactcg agcaccacca
     3781 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     3841 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     3901 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     3961 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4021 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4081 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4141 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4201 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4261 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4321 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4381 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4441 tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca
     4501 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     4561 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     4621 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     4681 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     4741 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     4801 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     4861 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     4921 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     4981 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     5041 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     5101 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     5161 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     5221 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     5281 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     5341 tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5401 cgtagaaaag atcaaaggat cttcttgata tccttttttt ctgcgcgtaa tctgctgctt
     5461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6181 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6241 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6301 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     6361 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6421 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     6481 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     6541 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     6601 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     6661 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     6721 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     6781 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     6841 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     6901 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     6961 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     7021 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     7081 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     7141 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     7201 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     7261 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     7321 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     7381 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     7441 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7501 gcgcggggag aggcggtttg cgtattgggc gccagggtgg ttttt
//
