LOCUS       sequence-274861-        7698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express FL hnRNPF with N-terminal maltose binding protein tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7698)
  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 7698)
  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_FL
FEATURES             Location/Qualifiers
     source          1..7698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..94
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     178..197
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        178..196
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    197..221
                     /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             236..258
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             283..300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             313..1410
                     /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      1267..1276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1384..1407
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1429..1453
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1462..1482
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2740..2757
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2810..2828)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2824..2871
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2908..3363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2995..3014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3205..3226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3455..4270)
                     /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(3455..4270)
                     /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     4178..4197
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4262..4281
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4392..4980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4881..4900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5134..5151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5166..5305
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5231..5250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5391..5413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5407..5598)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6407..7489)
                     /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     7451..7470
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7490..7567)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
       61 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
      121 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
      181 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
      241 ttaactttaa gaaggagata taccatggga tctgataaaa ttcatcatca tcatcatcac
      301 agcagcggta ccaaaatcga agaaggtaaa ctggtaatct ggattaacgg cgataaaggc
      361 tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat taaagtcacc
      421 gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc
      481 cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc tggcctgttg
      541 gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac ctgggatgcc
      601 gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt
      661 tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc ggcgctggat
      721 aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga accgtacttc
      781 acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa cggcaagtac
      841 gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac cttcctggtt
      901 gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc agaagctgcc
      961 tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc caacatcgac
     1021 accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca accatccaaa
     1081 ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa agagctggca
     1141 aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt taataaagac
     1201 aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggcgaa agatccacgt
     1261 attgccgcca ccatggaaaa cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg
     1321 tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact
     1381 gtcgatgaag ccctgaaaga cgcgcagact aattcgggat ctgatatcac aagtttgtac
     1441 aaaaaagctg aaggcggtac cgagaacctg tacttccagg gtcatatgat gctgggtcct
     1501 gagggtggcg agggtttcgt tgtcaagttg cgcggtctgc cgtggtcttg ctctgttgaa
     1561 gatgtccaga acttcctgtc ggattgtact atccacgatg gcgctgccgg tgtgcacttc
     1621 atctataccc gtgagggtcg ccagagcggt gaggccttcg ttgagctggg ttccgaagat
     1681 gacgtcaaaa tggcgctgaa aaaagaccgt gaatctatgg gccaccgtta tattgaagtg
     1741 ttcaaaagcc accgtaccga gatggactgg gttctgaagc acagcggtcc gaacagcgca
     1801 gatagcgcga atgatggttt tgttcgtctg cgtggcctgc cttttggttg caccaaagaa
     1861 gaaattgtcc aatttttctc gggtctggaa atcgttccga atggcattac gctgccggtt
     1921 gatccggaag gcaagattac cggtgaggca ttcgtgcaat ttgctagcca agaactggca
     1981 gagaaagcac tgggcaagca caaggaacgt attggtcacc gctacatcga agtattcaag
     2041 agcagccaag aggaagtgcg cagctatagc gatccgccgc tgaagtttat gtccgtgcag
     2101 cgtccgggtc catacgaccg cccgggcacc gcccgtcgtt atatcggtat cgtgaaacag
     2161 gccggcttgg agcgcatgcg cccgggtgcg tatagcacgg gctacggtgg ctacgaagag
     2221 tactctggcc tgagcgacgg ctatggtttt acgacggacc tgttcggtcg tgacttgagc
     2281 tattgcctga gcggtatgta cgaccaccgt tatggtgatt ccgagtttac cgtccagtct
     2341 accaccggtc attgtgttca catgcgtggt ctgccataca aggcaaccga gaatgacatc
     2401 tacaacttct tcagcccgct gaatccggtg cgtgtccaca tcgaaattgg tccggatggt
     2461 cgtgtcacgg gtgaggctga tgtcgaattt gcgactcatg aagaagccgt ggcggcgatg
     2521 agcaaagacc gtgcgaacat gcagcatcgc tacattgagt tgtttctgaa cagcaccacg
     2581 ggcgcaagca acggtgcgta ctccagccaa gttatgcaag gcatgggtgt gagcgcggca
     2641 caggcgacgt acagcggcct ggagagccag tccgttagcg gctgctatgg tgcgggctat
     2701 agcggtcaga acagcatggg cggctatgac tgactcgagc accaccacca ccaccactga
     2761 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2821 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     2881 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     2941 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3001 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3061 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3121 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3181 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3241 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3301 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3361 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3421 acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat
     3481 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     3541 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     3601 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     3661 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
     3721 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     3781 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     3841 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     3901 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     3961 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     4021 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     4081 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     4141 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     4201 ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa
     4261 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     4321 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4381 aaaggatctt cttgatatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4441 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4501 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     4561 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4621 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4681 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4741 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4801 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4861 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4921 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4981 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5041 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5101 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5161 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     5221 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     5281 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     5341 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5401 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     5461 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     5521 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     5581 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     5641 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     5701 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     5761 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     5821 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     5881 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     5941 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     6001 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     6061 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     6121 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     6181 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     6241 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     6301 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     6361 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     6541 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     6601 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     6661 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     6721 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     6781 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     6841 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     6901 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     6961 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     7021 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     7081 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     7141 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     7201 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     7261 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     7321 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     7381 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     7441 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     7501 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     7561 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     7621 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     7681 cccaacagtc ccccggcc
//
