LOCUS       sequence-201655-        7476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express MBP-tagged full length hnRNPA2 P298L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7476)
  AUTHORS   Ryan VH, Dignon GL, Zerze GH, Chabata CV, Silva R, Conicella AE, 
            Amaya J, Burke KA, Mittal J, Fawzi NL
  TITLE     Mechanistic View of hnRNPA2 Low-Complexity Domain Structure, 
            Interactions, and Phase Separation Altered by Mutation and Arginine 
            Methylation.
  JOURNAL   Mol Cell. 2018 Jan 17. pii: S1097-2765(17)30979-6. doi: 
            10.1016/j.molcel.2017.12.022.
  PUBMED    29358076
REFERENCE   2  (bases 1 to 7476)
  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: "Mol Cell. 
            2018 Jan 17. pii: S1097-2765(17)30979-6. doi: 
            10.1016/j.molcel.2017.12.022."
COMMENT     Sequence Label: MBP-hnRNPA2_FL_P298L
FEATURES             Location/Qualifiers
     source          1..7476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             190..207
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(260..278)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      274..321
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      358..813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(445..464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     655..676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(905..1720)
                     /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(905..1720)
                     /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     1628..1647
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1712..1731
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1842..2430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2331..2350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2584..2601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2616..2755
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2681..2700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2841..2863
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2857..3048)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3857..4939)
                     /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     4901..4920
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4940..5017)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5223..5242
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5326..5345
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5326..5344
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5345..5369
                     /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             5384..5406
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5431..5448
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5461..6558
                     /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      6415..6424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     6532..6555
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    6577..6601
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6610..6630
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 aactatggta gcggcaatta caatgatttt ggcaactaca accagcaact gagcaattac
       61 ggcccgatga aatccggcaa ttttggcggt tcccgtaata tgggtggtcc gtatggtggc
      121 ggtaactacg gcccgggtgg tagcggtggt tctggcggct atggtggccg cagccgttac
      181 taactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
      241 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      301 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
      361 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      421 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      481 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      541 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      601 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
      661 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
      721 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
      781 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
      841 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
      901 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
      961 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1021 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1081 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1141 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     1201 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1261 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     1321 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     1381 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     1441 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     1501 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     1561 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     1621 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     1681 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     1741 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     1801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgatatcc tttttttctg
     1861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1981 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2581 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     2641 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     2701 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     2761 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2821 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2881 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     2941 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3001 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     3061 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     3121 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     3181 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     3241 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     3301 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     3361 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     3421 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     3481 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     3541 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     3601 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     3661 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     3721 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     3781 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     3841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3901 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     3961 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     4021 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     4081 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     4141 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     4201 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     4261 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     4321 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     4381 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     4441 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     4501 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     4561 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     4621 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     4681 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     4741 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     4801 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     4861 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     4921 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     4981 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     5041 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     5101 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     5161 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     5221 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     5281 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     5341 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     5401 aggagatata ccatgggatc tgataaaatt catcatcatc atcatcacag cagcggtacc
     5461 aaaatcgaag aaggtaaact ggtaatctgg attaacggcg ataaaggcta taacggtctc
     5521 gctgaagtcg gtaagaaatt cgagaaagat accggaatta aagtcaccgt tgagcatccg
     5581 gataaactgg aagagaaatt cccacaggtt gcggcaactg gcgatggccc tgacattatc
     5641 ttctgggcac acgaccgctt tggtggctac gctcaatctg gcctgttggc tgaaatcacc
     5701 ccggacaaag cgttccagga caagctgtat ccgtttacct gggatgccgt acgttacaac
     5761 ggcaagctga ttgcttaccc gatcgctgtt gaagcgttat cgctgattta taacaaagat
     5821 ctgctgccga acccgccaaa aacctgggaa gagatcccgg cgctggataa agaactgaaa
     5881 gcgaaaggta agagcgcgct gatgttcaac ctgcaagaac cgtacttcac ctggccgctg
     5941 attgctgctg acgggggtta tgcgttcaag tatgaaaacg gcaagtacga cattaaagac
     6001 gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa
     6061 aacaaacaca tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc
     6121 gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg
     6181 aattatggtg taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc
     6241 gtgctgagcg caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc
     6301 gaaaactatc tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt
     6361 gccgtagcgc tgaagtctta cgaggaagag ttggcgaaag atccacgtat tgccgccacc
     6421 atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg
     6481 tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc
     6541 ctgaaagacg cgcagactaa ttcgggatct gatatcacaa gtttgtacaa aaaagctgaa
     6601 ggcggtaccg agaacctgta cttccagggt catatggaac gtgaaaaaga gcagtttcgc
     6661 aagctgttta tcggtggcct gagcttcgaa actaccgaag agagcctgcg caactattat
     6721 gagcagtggg gtaaactgac ggattgcgtc gtgatgcgtg acccggcgag caagcgcagc
     6781 cgtggcttcg gtttcgtcac cttctccagc atggcagagg ttgacgccgc tatggcagcc
     6841 cgtcctcaca gcattgacgg ccgtgtggtt gagccgaagc gtgcggtcgc tcgtgaagag
     6901 tcgggtaagc cgggtgctca cgtcaccgtc aaaaagctgt tcgtgggtgg tatcaaggaa
     6961 gataccgagg aacaccactt gcgcgactac ttcgaggaat atggcaaaat cgataccatt
     7021 gagatcatta cggaccgtca aagcggcaaa aagcgtggtt tcggctttgt tacgtttgac
     7081 gatcacgacc cggttgacaa aatcgtgctg caaaagtacc acaccattaa cggccacaac
     7141 gccgaggtgc gtaaagcgct gagccgtcaa gagatgcaag aagtgcagtc ctctcgcagc
     7201 ggtcgtggtg gcaactttgg tttcggtgat agccgcggtg gtggcggcaa cttcggtccg
     7261 ggtccgggca gcaatttccg tggtggttcc gacggttacg gtagcggtcg cggctttggc
     7321 gatggctata atggttacgg tggtggtccg ggcggcggca atttcggtgg tagcccgggt
     7381 tacggcggtg gtcgcggcgg ttatggcggt ggcggtccag gctacggtaa ccagggtggt
     7441 ggttatggtg gtggctacga taactatggc ggtggt
//
