LOCUS       sequence-273912-        6305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of HNRNPH2 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6305)
  AUTHORS   Dominguez D, Freese P, Alexis MS, Su A, Hochman M, Palden T, Bazile 
            C, Lambert NJ, Van Nostrand EL, Pratt GA, Yeo GW, Graveley BR, Burge
            CB
  TITLE     Sequence, Structure, and Context Preferences of Human RNA Binding 
            Proteins.
  JOURNAL   Mol Cell. 2018 Jun 7;70(5):854-867.e9. doi: 
            10.1016/j.molcel.2018.05.001. Epub 2018 Jun 7.
  PUBMED    29883606
REFERENCE   2  (bases 1 to 6305)
  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"; doi: 
            "10.1016/j.molcel.2018.05.001"; journalName: "Mol Cell"; date: 
            "2018-06-7- 7"; volume: "70"; issue: "5"; pages: "854-867.e9"
COMMENT     Sequence Label: HNRNPH2_3xRRM_pGEX
FEATURES             Location/Qualifiers
     source          1..6305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..723
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             733..846
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     primer_bind     complement(2137..2159)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2304..2322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2390..2494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2495..2563,2564..3355)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2713..2732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3526..4114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4015..4034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4268..4285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4358..4435
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4436..5518
                     /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(4455..4474)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5567..5572,5573..5590,5591..5597)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5605..5621
                     /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)."
     primer_bind     5629..5645
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5629..5645
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5641..6168
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5661..5678)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5661..5677)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5670..5692)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6270..6275,6276..6291,6292..6298)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggaagttct gttccagggg
      721 cccctgggat taatggacga aaaaaccacc ggttggcgtg gtggtcacgt tgttgaaggt
      781 ctggctggtg aactggaaca gctgcgtgct cgtctggaac accacccgca gggtcagcgt
      841 gaaccgggat ccatgatgct gagcacggaa ggcagggagg ggttcgtggt gaaggtcagg
      901 ggcctaccct ggtcctgctc agccgatgaa gtgatgcgct tcttctctga ttgcaagatc
      961 caaaatggca catcaggtat tcgtttcatc tacaccagag aaggcagacc aagtggtgaa
     1021 gcatttgttg aacttgaatc tgaagaggaa gtgaaattgg ctttgaagaa ggacagagaa
     1081 accatgggac acagatacgt tgaagtattc aagtctaaca gtgttgaaat ggattgggtg
     1141 ttgaagcata caggtccgaa tagccctgat actgccaacg atggcttcgt ccggcttaga
     1201 ggactcccat ttggctgtag caaggaagag attgttcagt tcttttcagg tttggaaatt
     1261 gtgccaaatg ggatgacact gccagtggac tttcaggggc gaagcacagg ggaagccttt
     1321 gtgcagtttg cttcacagga gatagctgag aaggccttaa agaaacacaa ggaaagaata
     1381 gggcacaggt acattgagat cttcaagagt agccgagctg aagttcgaac ccactatgat
     1441 ccccctcgaa agctcatggc tatgcagcgg ccaggtccct atgataggcc gggggctggc
     1501 agagggtata atagcattgg cagaggagct gggtttgaaa ggatgaggcg tggtgcctat
     1561 ggtggagggt atggaggcta tgatgactat ggtggctata atgatggata tggctttggg
     1621 tctgatagat ttggaagaga cctcaattac tgtttttcag gaatgtctga tcatagatac
     1681 ggagatggtg ggtccagttt ccagagcacc acagggcact gtgtacacat gagggggtta
     1741 ccttacagag ccactgagaa tgatatttat aatttcttct cacctcttaa tcccatgaga
     1801 gtacatattg aaattggacc cgatggcaga gttaccggtg aggcagatgt tgaatttgct
     1861 actcatgaag atgctgtggc agctatggca aaagacaaag ctaatatgca acacagatat
     1921 gtggagctct tcttaaattc tactgcagga acaagtgggg gtgcttacga tcacagctat
     1981 gtagaacttt ttttgaattc tacagcaggg gcaagtggtg gcgcttatgg tagccaaatg
     2041 atgggaggga tgggcttatc caaccagtct agttaggcgg ccgcatcgtg actgactgac
     2101 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2161 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2221 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     2281 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2341 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2401 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2461 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2521 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2581 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2641 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2701 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     2761 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2821 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2881 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2941 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3001 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     3061 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3121 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3181 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3241 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3301 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3361 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3421 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3481 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3541 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3601 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3661 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3721 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3781 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3841 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3901 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3961 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4021 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4081 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4141 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4201 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4261 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4321 tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat ggcgcaaaac
     4381 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     4441 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     4501 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     4561 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     4621 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     4681 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     4741 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     4801 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     4861 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     4921 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     4981 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     5041 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     5101 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     5161 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     5221 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     5281 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     5341 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     5401 actggtgaaa agaaaaacca cgctagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5461 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5521 gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5581 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5641 atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5701 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5761 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     5821 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     5881 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     5941 tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     6001 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     6061 cgaattattt ttgatggcgt tggaattagc ttatcgactg cacggtgcac caatgcttct
     6121 ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa
     6181 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     6241 ggttctggca aatattctga aatgagctgt tgacaattaa tcatcggctc gtataatgtg
     6301 tggaa
//
