LOCUS       sequence-307928-        5028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express His tagged hnRNPA2 RRMs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5028)
  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 5028)
  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: hnRNPA2_RRMs
FEATURES             Location/Qualifiers
     source          1..5028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     208..227
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        208..226
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    227..251
                     /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             312..329
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             330..350
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      1390..1437
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1660..1679)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1746..1837
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1838..2647
                     /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             2730..3812
                     /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(2749..2768)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3898..4486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4387..4406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4640..4657
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4755..4772
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4756..4772
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attatttatg attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat
       61 aaatctcgaa aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg
      121 ataatacaaa atataataca aactataaga tgttatcagt atttattatg catttagaat
      181 aaattttgtg tcgcccttcc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      241 ataacaattc ccctctagaa ataattttgt ttaacttttg aaggagatat accatgggta
      301 gcgacaagat tcaccaccat catcatcatg agaatttgta ctttcagggc catatggaac
      361 gtgaaaaaga gcagtttcgc aagctgttta tcggtggcct gagcttcgaa actaccgaag
      421 agagcctgcg caactattat gagcagtggg gtaaactgac ggattgcgtc gtgatgcgtg
      481 acccggcgag caagcgcagc cgtggcttcg gtttcgtcac cttctccagc atggcagagg
      541 ttgacgccgc tatggcagcc cgtcctcaca gcattgacgg ccgtgtggtt gagccgaagc
      601 gtgcggtcgc tcgtgaagag tcgggtaagc cgggtgctca cgtcaccgtc aaaaagctgt
      661 tcgtgggtgg tatcaaggaa gataccgagg aacaccactt gcgcgactac ttcgaggaat
      721 atggcaaaat cgataccatt gagatcatta cggaccgtca aagcggcaaa aagcgtggtt
      781 tcggctttgt tacgtttgac gatcacgacc cggttgacaa aatcgtgctg caaaagtacc
      841 acaccattaa cggccacaac gccgaggtgc gtaaagcgct gagccgtcaa gagatgcaag
      901 aagtgcagtc ctctcgcagc tagcgtggtg gcaactttgg tttcggtgat agccgcggtg
      961 gtggcggcaa cttcggtccg ggtccgggca gcaatttccg tggtggttcc gacggttacg
     1021 gtagcggtcg cggctttggc gatggctata atggttacgg tggtggtccg ggcggcggca
     1081 atttcggtgg tagcccgggt tacggcggtg gtcgcggcgg ttatggcggt ggcggtccag
     1141 gctacggtaa ccagggtggt ggttatggtg gtggctacga taactatggc ggtggtaact
     1201 atggtagcgg caattacaat gattttggca actacaacca gcaaccgagc aattacggcc
     1261 cgatgaaatc cggcaatttt ggcggttccc gtaatatggg tggtccgtat ggtggcggta
     1321 actacggccc gggtggtagc ggtggttctg gcggctatgg tggccgcagc cgttactaac
     1381 tcgagccccc tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgccc
     1441 ctgagacgcg tcaatcgagt tcgtacctaa gggcgacacc ccctaattag cccgggcgaa
     1501 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     1561 gcatggggag tccccacact accatcggcg ctacggcgtt tcacttctga gttcggcatg
     1621 gggtcaggtg ggaccaccgc gctactgccg ccaggcaaac aaggggtgtt atgagccata
     1681 ttcaggtata aatgggctcg cgataatgtt cagaattggt taattggttg taacactgac
     1741 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1801 ctgataaatg cttcaataat attgaaaaag gaagaatatg agccatattc aacgggaaac
     1861 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1921 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1981 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2041 cagactaaac tggctgacgg aatttatgcc acttccgacc atcaagcatt ttatccgtac
     2101 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcgt tccaggtatt
     2161 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2221 gttgcactcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc
     2281 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2341 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2401 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2461 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2521 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2581 atatggtatt gataatcctg atatgaataa attgcaattt catttgatgc tcgatgagtt
     2641 tttctaagcg gcgcgccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     2701 cggaagagag tcaattcagg gtggtgaata tgaaaccagt aacgttatac gatgtcgcag
     2761 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     2821 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     2881 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     2941 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     3001 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     3061 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     3121 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     3181 ctgaccagac acccatcaac agtattattt tctcccatga ggacggtacg cgactgggcg
     3241 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     3301 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     3361 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     3421 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     3481 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     3541 tgggatacga cgataccgaa gatagctcat gttatatccc gccgttaacc accatcaaac
     3601 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     3661 aggcggtgaa gggcaatcag ctgttgccag tctcactggt gaaaagaaaa accaccctgg
     3721 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3781 gacaggtttc ccgactggaa agcgggcagt gactcatgac caaaatccct taacgtgagt
     3841 tacgcgcgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa
     4081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4561 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4621 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga
     4681 actgccaggc atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca
     4741 aactctttct gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct
     4801 gataacgagt aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta
     4861 tggggggagt ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct
     4921 tgatatatga gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga
     4981 tacgttgctt tttcgattga tgaacaccta taattaaact attcatct
//
