LOCUS       sequence-273982-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of RBM23 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  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 5942)
  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: RBM23_2xRRM_pGEX
FEATURES             Location/Qualifiers
     source          1..5942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(165..170,171..188,189..195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    203..219
                     /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     227..243
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     227..243
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             239..766
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(259..276)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(259..275)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(268..290)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(868..873,874..889,890..896)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    904..920
                     /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             943..1596
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1554..1576
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1603..1626
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1636..1749
                     /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(2677..2699)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2844..2862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2930..3034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3035..3103,3104..3895)
                     /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(3253..3272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4066..4654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4555..4574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4808..4825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4898..4975
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4976..5942,1..116)
                     /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(4995..5014)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggtgaaaag aaaaaccacg ctagcgccca atacgcaaac cgcctctccc cgcgcgttgg
       61 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
      121 aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt
      181 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
      241 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
      301 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      361 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
      421 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
      481 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
      541 caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
      601 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
      661 aattattttt gatggcgttg gaattagctt atcgactgca cggtgcacca atgcttctgg
      721 cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt
      781 cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg
      841 ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg
      901 gaattgtgag cggataacaa tttcacacag gaaacagtat tcatgtcccc tatactaggt
      961 tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa
     1021 aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt
     1081 gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca
     1141 cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca
     1201 aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt
     1261 tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta
     1321 cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat
     1381 catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac
     1441 ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc
     1501 ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg
     1561 caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt tctgttccag
     1621 gggcccctgg gattaatgga cgaaaaaacc accggttggc gtggtggtca cgttgttgaa
     1681 ggtctggctg gtgaactgga acagctgcgt gctcgtctgg aacaccaccc gcagggtcag
     1741 cgtgaaccgg gatcccaccg tagccgtagc tgggatcgtc gacatggtag tgagtcgcga
     1801 agtcgggacc atcgtcgtga ggatcgtgtg cattacagga gtcctccact tgccactggt
     1861 tataggtatg gacacagtaa gagtcctcat ttcagagaga agagcccagt cagggagcca
     1921 gttgataatc tgagtcctga ggagcgtgat gcccgcacag ttttctgtat gcagttagct
     1981 gcccgaattc ggcctcgaga tctggaggac tttttctctg ctgtaggcaa ggttcgcgat
     2041 gtacgtatca tctcagatcg gaactcacgt cgttctaagg gcattgccta cgtggaattc
     2101 tgtgaaatcc agtctgtgcc actggccatt gggctgactg ggcagcggtt gctgggagtg
     2161 cctatcattg tacaggcttc acaggcagag aaaaaccgac tggcagccat ggccaacaac
     2221 ctgcaaaagg gcaatggtgg accaatgcgc ctctatgtgg gttccctgca cttcaatatc
     2281 actgaagaca tgctccgggg catctttgag ccctttggta aaattgataa tattgtcctg
     2341 atgaaggact cagatacagg ccgctctaaa ggttatggtt tcatcacgtt ctctgattct
     2401 gagtgtgccc ggcgggccct ggaacagttg aatgggtttg agcttgctgg tcgacctatg
     2461 agggttggcc atgtgactga gcgactggat ggtggcacag acatcacttt tcctgatggg
     2521 gaccaggagc tggatctggg atcagcaggt ggacgttttc agctcatggc aaagctggca
     2581 gaaggcgctg gaatccaact gccaagcact gcttaggcgg ccgcatcgtg actgactgac
     2641 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2701 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2761 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     2821 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2881 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2941 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3001 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3061 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3121 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3181 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3241 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     3301 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3361 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3421 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3481 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3541 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     3601 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3661 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3721 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3781 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3841 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3901 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3961 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4081 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4141 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4201 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4261 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4321 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4381 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4441 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4501 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4561 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4621 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4681 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4741 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4801 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4861 tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat ggcgcaaaac
     4921 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     4981 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     5041 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     5101 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     5161 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     5221 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     5281 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     5341 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     5401 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     5461 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     5521 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     5581 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     5641 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     5701 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     5761 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     5821 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     5881 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     5941 ac
//
