LOCUS       sequence-274920-        6168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FL hnRNPA2 P298L with a C-terminal MBP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  AUTHORS   Ryan VH, Watters S, Amaya J, Khatiwada B, Venditti V, Naik MT, Fawzi
            NL
  TITLE     Weak binding to the A2RE RNA rigidifies hnRNPA2 RRMs and reduces 
            liquid-liquid phase separation and aggregation.
  JOURNAL   Nucleic Acids Res. 2020 Sep 1. pii: 5900113. doi: 
            10.1093/nar/gkaa710.
  PUBMED    32870271
REFERENCE   2  (bases 1 to 6168)
  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: "Nucleic 
            Acids Res. 2020 Sep 1. pii: 5900113. doi: 10.1093/nar/gkaa710."
COMMENT     Sequence Label: pJ4M_hnRNPA2_FL_P298L
FEATURES             Location/Qualifiers
     source          1..6168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..175
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        156..174
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    175..199
                     /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)."
     CDS             1274..1294
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1304..2401
                     /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      2258..2267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2375..2398
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    2420..2444
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2453..2470
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2478..2525
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2748..2767)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2834..2925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2926..3735
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3818..4900
                     /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     complement(3837..3856)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      4986..5574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5475..5494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5728..5745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5843..5860
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5844..5860
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca aaattataca
       61 tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat ttattatgca
      121 tttagaataa attttgtgtc gcccttccgc gaaattaata cgactcacta taggggaatt
      181 gtgagcggat aacaattccc ctctagaaat aattttgttt aacttttgaa ggagatatac
      241 ccatatggaa cgtgaaaaag agcagtttcg caagctgttt atcggtggcc tgagcttcga
      301 aactaccgaa gagagcctgc gcaactatta tgagcagtgg ggtaaactga cggattgcgt
      361 cgtgatgcgt gacccggcga gcaagcgcag ccgtggcttc ggtttcgtca ccttctccag
      421 catggcagag gttgacgccg ctatggcagc ccgtcctcac agcattgacg gccgtgtggt
      481 tgagccgaag cgtgcggtcg ctcgtgaaga gtcgggtaag ccgggtgctc acgtcaccgt
      541 caaaaagctg ttcgtgggtg gtatcaagga agataccgag gaacaccact tgcgcgacta
      601 cttcgaggaa tatggcaaaa tcgataccat tgagatcatt acggaccgtc aaagcggcaa
      661 aaagcgtggt ttcggctttg ttacgtttga cgatcacgac ccggttgaca aaatcgtgct
      721 gcaaaagtac cacaccatta acggccacaa cgccgaggtg cgtaaagcgc tgagccgtca
      781 agagatgcaa gaagtgcagt cctctcgcag cggtcgtggt ggcaactttg gtttcggtga
      841 tagccgcggt ggtggcggca acttcggtcc gggtccgggc agcaatttcc gtggtggttc
      901 cgacggttac ggtagcggtc gcggctttgg cgatggctat aatggttacg gtggtggtcc
      961 gggcggcggc aatttcggtg gtagcccggg ttacggcggt ggtcgcggcg gttatggcgg
     1021 tggcggtcca ggctacggta accagggtgg tggttatggt ggtggctacg ataactatgg
     1081 cggtggtaac tatggtagcg gcaattacaa tgattttggc aactacaacc agcaactgag
     1141 caattacggc ccgatgaaat ccggcaattt tggcggttcc cgtaatatgg gtggtccgta
     1201 tggtggcggt aactacggcc cgggtggtag cggtggttct ggcggctatg gtggccgcag
     1261 ccgttacctc gaggagaacc tgtactttca gggtagcggt accaaaatcg aagaaggtaa
     1321 actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa
     1381 attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa
     1441 attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg
     1501 ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca
     1561 ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta
     1621 cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc
     1681 aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc
     1741 gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg
     1801 ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc
     1861 tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc
     1921 agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat
     1981 caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt
     2041 actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat
     2101 taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac
     2161 tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc
     2221 ttacgaggaa gagttggcga aagatccacg tattgccgcc accatggaaa acgcccagaa
     2281 aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc
     2341 ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac
     2401 taattcggga tctgatatca caagtttgta caaaaaagct gaaggcggta cccatcacca
     2461 tcaccatcat tgacccccta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2521 ttttgcccct gagacgcgtc aatcgagttc gtacctaagg gcgacacccc ctaattagcc
     2581 cgggcgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggcagttcc
     2641 ctactctcgc atggggagtc cccacactac catcggcgct acggcgtttc acttctgagt
     2701 tcggcatggg gtcaggtggg accaccgcgc tactgccgcc aggcaaacaa ggggtgttat
     2761 gagccatatt caggtataaa tgggctcgcg ataatgttca gaattggtta attggttgta
     2821 acactgaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2881 caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag ccatattcaa
     2941 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     3001 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     3061 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     3121 gagatggtca gactaaactg gctgacggaa tttatgccac ttccgaccat caagcatttt
     3181 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcgttc
     3241 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     3301 ctgcgccggt tgcactcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     3361 cgcctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     3421 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     3481 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3541 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     3601 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     3661 tttcaaaaat atggtattga taatcctgat atgaataaat tgcaatttca tttgatgctc
     3721 gatgagtttt tctaagcggc gcgccatcga atggcgcaaa acctttcgcg gtatggcatg
     3781 atagcgcccg gaagagagtc aattcagggt ggtgaatatg aaaccagtaa cgttatacga
     3841 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     3901 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     3961 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     4021 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     4081 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     4141 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     4201 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     4261 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgagg acggtacgcg
     4321 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     4381 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     4441 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     4501 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     4561 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     4621 ctcggtagtg ggatacgacg ataccgaaga tagctcatgt tatatcccgc cgttaaccac
     4681 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     4741 tcagggccag gcggtgaagg gcaatcagct gttgccagtc tcactggtga aaagaaaaac
     4801 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4861 gctggcacga caggtttccc gactggaaag cgggcagtga ctcatgacca aaatccctta
     4921 acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5101 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttagcccac
     5161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5281 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5641 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5701 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag
     5761 agtagggaac tgccaggcat caaactaagc agaaggcccc tgacggatgg cctttttgcg
     5821 tttctacaaa ctctttctgt gttgtaaaac gacggccagt cttaagctcg ggccccctgg
     5881 gcggttctga taacgagtaa tcgttaatcc gcaaataacg taaaaacccg cttcggcggg
     5941 tttttttatg gggggagttt agggaaagag catttgtcag aatatttaag ggcgcctgtc
     6001 actttgcttg atatatgaga attatttaac cttataaatg agaaaaaagc aacgcacttt
     6061 aaataagata cgttgctttt tcgattgatg aacacctata attaaactat tcatctatta
     6121 tttatgattt tttgtatata caatatttct agtttgttaa agagaatt
//
