LOCUS       sequence-189359-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged hnRNPA2 AA 190-341 with D290V and S329C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  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 5751)
  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: hnRNPA2_LC_D290V_S329C
FEATURES             Location/Qualifiers
     source          1..5751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             106..123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             124..144
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             616..633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(686..704)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      700..747
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      784..1239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(871..890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1081..1102
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1331..2146)
                     /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(1331..2146)
                     /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     2054..2073
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2138..2157
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2268..2856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2757..2776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3010..3027
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3042..3181
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3107..3126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3267..3289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3283..3474)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4283..5365)
                     /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     5327..5346
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5366..5443)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5649..5668
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggctctgata aaattcatca tcatcatcat
      121 cacgaaaatc tttattttca aggtcatatg ggtcgtggtg gcaactttgg tttcggtgat
      181 agccgcggtg gtggcggcaa cttcggtccg ggtccgggca gcaatttccg tggtggttcc
      241 gacggttacg gtagcggtcg cggctttggc gatggctata atggttacgg tggtggtccg
      301 ggcggcggca atttcggtgg tagcccgggt tacggcggtg gtcgcggcgg ttatggcggt
      361 ggcggtccag gctacggtaa ccagggtggt ggttatggtg gtggctacga taactatggc
      421 ggtggtaact atggtagcgg caattacaat gtttttggca actacaacca gcaaccgagc
      481 aattacggcc cgatgaaatc cggcaatttt ggcggttccc gtaatatggg tggtccgtat
      541 ggtggcggta actacggccc gggtggttgc ggtggttctg gcggctatgg tggccgcagc
      601 cgttactaac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
      661 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      721 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
      781 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      841 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      901 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      961 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1021 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1081 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1141 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1201 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     1261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1321 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     1381 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1441 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1501 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1561 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     1621 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1681 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     1741 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1801 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     1861 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1921 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1981 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     2041 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     2101 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     2161 tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt
     2221 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2281 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2341 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2401 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2461 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2521 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2581 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2641 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2701 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2761 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2821 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2881 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2941 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3001 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3061 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     3121 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     3181 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3241 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3301 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     3361 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3421 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     3481 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     3541 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     3601 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     3661 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     3721 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     3781 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     3841 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     3901 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat
     3961 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     4021 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     4081 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     4141 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     4201 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     4261 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4321 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     4381 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     4441 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     4501 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     4561 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     4621 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4681 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4741 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     4801 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     4861 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     4921 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     4981 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     5041 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     5101 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     5161 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     5221 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     5281 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     5341 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     5401 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     5461 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     5521 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     5581 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     5641 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     5701 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
