LOCUS       sequence-275068-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of RBM24 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  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 5504)
  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: RBM24_RRM_pGEX
FEATURES             Location/Qualifiers
     source          1..5504
                     /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(1336..1358)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1503..1521)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1589..1693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1694..1762,1763..2554)
                     /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(1912..1931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2725..3313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3214..3233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3467..3484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3557..3634
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3635..4717
                     /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(3654..3673)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4766..4771,4772..4789,4790..4796)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4804..4820
                     /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     4828..4844
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4828..4844
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4840..5367
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4860..4877)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4860..4876)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4869..4891)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5469..5474,5475..5490,5491..5497)
                     /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 ccatgcacac gacccagaag gacacgacgt acaccaagat cttcgtcggg
      901 gggctgccct accacaccac cgacgccagc ctgcgcaagt acttcgaggt cttcggcgag
      961 atcgaggagg cggtggtcat caccgaccgg cagacgggca agtcccgggg ctatggattt
     1021 gtcaccatgg ctgaccgggc tgctgccgaa agggcctgca aggatcccaa tcccatcatt
     1081 gatggcagaa aggccaacgt gaacctggca tacttaggag caaaaccaag gatcatgcaa
     1141 ccaggttttg cctttggtgt tcaacaactt catccagccc ttatacaaag acctttcggg
     1201 atacctgccc actatgtcta tccgcaggct tttgtgcagc cgggagtggt cattccacac
     1261 gtccagccga catgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg
     1321 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1381 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1441 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa
     1501 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1561 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1621 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1681 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1741 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1801 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1861 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1921 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     1981 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2041 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2101 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2161 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2221 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2521 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2581 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2641 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3421 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3481 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3541 acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     3601 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     3661 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     3721 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     3781 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3841 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     3901 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     3961 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     4021 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     4081 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     4141 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     4201 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     4261 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     4321 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     4381 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     4441 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     4501 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     4561 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     4621 gctagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4681 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     4741 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     4801 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca
     4861 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4921 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4981 ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca
     5041 gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc
     5101 ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc
     5161 attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca
     5221 tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt
     5281 ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
     5341 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
     5401 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
     5461 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaa
//
