LOCUS       sequence-274842-        6168 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express FL hnRNPA2 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 Apr 14, 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_WT
FEATURES             Location/Qualifiers
     source          1..6168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     270..289
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        270..288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    289..313
                     /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             1388..1408
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1418..2515
                     /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      2372..2381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2489..2512
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    2534..2558
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2567..2584
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2592..2639
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2862..2881)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2948..3039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3040..3849
                     /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             3932..5014
                     /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(3951..3970)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      5100..5688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5589..5608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5842..5859
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5957..5974
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5958..5974
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cactttaaat aagatacgtt gctttttcga ttgatgaaca cctataatta aactattcat
       61 ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag aattaagaaa
      121 ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta tacatgtcaa
      181 cgataataca aaatataata caaactataa gatgttatca gtatttatta tgcatttaga
      241 ataaattttg tgtcgccctt ccgcgaaatt aatacgactc actatagggg aattgtgagc
      301 ggataacaat tcccctctag aaataatttt gtttaacttt tgaaggagat atacccatat
      361 ggaacgtgaa aaagagcagt ttcgcaagtt gtttatcggt ggcctgagct tcgaaactac
      421 cgaagagagc ctgcgcaact attatgagca gtggggtaaa ctgacggatt gcgtcgtgat
      481 gcgtgacccg gcgagcaagc gcagccgtgg cttcggtttc gtcaccttct ccagcatggc
      541 agaggttgac gccgctatgg cagcccgtcc tcacagcatt gacggccgtg tggttgagcc
      601 gaagcgtgcg gtcgctcgtg aagagtcggg taagccgggt gctcacgtca ccgtcaaaaa
      661 gctgttcgtg ggtggtatca aggaagatac cgaggaacac cacttgcgcg actacttcga
      721 ggaatatggc aaaatcgata ccattgagat cattacggac cgtcaaagcg gcaaaaagcg
      781 tggtttcggc tttgttacgt ttgacgatca cgacccggtt gacaaaatcg tgctgcaaaa
      841 gtaccacacc attaacggcc acaacgccga ggtgcgtaaa gcgctgagcc gtcaagagat
      901 gcaagaagtg cagtcctctc gcagcggtcg tggtggcaac tttggtttcg gtgatagccg
      961 cggtggtggc ggcaacttcg gtccgggtcc gggcagcaat ttccgtggtg gttccgacgg
     1021 ttacggtagc ggtcgcggct ttggcgatgg ctataatggt tacggtggtg gtccgggcgg
     1081 cggcaatttc ggtggtagcc cgggttacgg cggtggtcgc ggcggttatg gcggtggcgg
     1141 tccaggctac ggtaaccagg gtggtggtta tggtggtggc tacgataact atggcggtgg
     1201 taactatggt agcggcaatt acaatgattt tggcaactac aaccagcaac cgagcaatta
     1261 cggcccgatg aaatccggca attttggcgg ttcccgtaat atgggtggtc cgtatggtgg
     1321 cggtaactac ggcccgggtg gtagcggtgg ttctggcggc tatggtggcc gcagccgtta
     1381 cctcgaggag aacctgtact ttcagggtag cggtaccaaa atcgaagaag gtaaactggt
     1441 aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga
     1501 gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc
     1561 acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg
     1621 tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa
     1681 gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat
     1741 cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac
     1801 ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat
     1861 gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc
     1921 gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc
     1981 gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac
     2041 cgattactcc atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg
     2101 cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc
     2161 gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc
     2221 cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga
     2281 aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga
     2341 ggaagagttg gcgaaagatc cacgtattgc cgccaccatg gaaaacgccc agaaaggtga
     2401 aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat
     2461 caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaattc
     2521 gggatctgat atcacaagtt tgtacaaaaa agctgaaggc ggtacccatc accatcacca
     2581 tcattgaccc cctagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2641 ccctgagacg cgtcaatcga gttcgtacct aagggcgaca ccccctaatt agcccgggcg
     2701 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     2761 tcgcatgggg agtccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     2821 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa acaaggggtg ttatgagcca
     2881 tattcaggta taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg
     2941 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3001 ccctgataaa tgcttcaata atattgaaaa aggaagaata tgagccatat tcaacgggaa
     3061 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     3121 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     3181 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     3241 gtcagactaa actggctgac ggaatttatg ccacttccga ccatcaagca ttttatccgt
     3301 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc gttccaggta
     3361 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     3421 cggttgcact cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc
     3481 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3541 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3601 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3661 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3721 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3781 aaatatggta ttgataatcc tgatatgaat aaattgcaat ttcatttgat gctcgatgag
     3841 tttttctaag cggcgcgcca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     3901 cccggaagag agtcaattca gggtggtgaa tatgaaacca gtaacgttat acgatgtcgc
     3961 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     4021 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     4081 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     4141 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     4201 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     4261 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     4321 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     4381 ctctgaccag acacccatca acagtattat tttctcccat gaggacggta cgcgactggg
     4441 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     4501 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     4561 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     4621 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     4681 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     4741 agtgggatac gacgataccg aagatagctc atgttatatc ccgccgttaa ccaccatcaa
     4801 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     4861 ccaggcggtg aagggcaatc agctgttgcc agtctcactg gtgaaaagaa aaaccaccct
     4921 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4981 acgacaggtt tcccgactgg aaagcgggca gtgactcatg accaaaatcc cttaacgtga
     5041 gttacgcgcg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagc ccaccacttc
     5281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5641 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5701 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5761 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5821 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg cgagagtagg
     5881 gaactgccag gcatcaaact aagcagaagg cccctgacgg atggcctttt tgcgtttcta
     5941 caaactcttt ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc ctgggcggtt
     6001 ctgataacga gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg cgggtttttt
     6061 tatgggggga gtttagggaa agagcatttg tcagaatatt taagggcgcc tgtcactttg
     6121 cttgatatat gagaattatt taaccttata aatgagaaaa aagcaacg
//
