LOCUS       sequence-274423-        5624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of RBM15B with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5624)
  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 5624)
  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: RBM15B_3xRRM_pGEX
FEATURES             Location/Qualifiers
     source          1..5624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(481..486,487..504,505..511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    519..535
                     /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     543..559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     543..559
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             555..1082
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(575..592)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(575..591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(584..606)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1184..1189,1190..1205,1206..1212)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1220..1236
                     /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             1259..1912
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1870..1892
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1919..1942
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1952..2065
                     /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(2675..2697)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2842..2860)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2928..3032
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3033..3101,3102..3893)
                     /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(3251..3270)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4064..4652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4553..4572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4806..4823
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4896..4973
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4974..5624,1..432)
                     /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(4993..5012)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
       61 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
      121 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
      181 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
      241 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
      301 ctgttgcccg tctcactggt gaaaagaaaa accacgctag cgcccaatac gcaaaccgcc
      361 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      421 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc
      481 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      541 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
      601 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      661 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      721 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
      781 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
      841 acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc
      901 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
      961 aggaaggcca gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt
     1021 gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt
     1081 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
     1141 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg
     1201 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat
     1261 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
     1321 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
     1381 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
     1441 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
     1501 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
     1561 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
     1621 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
     1681 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
     1741 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
     1801 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
     1861 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct
     1921 ggaagttctg ttccaggggc ccctgggatt aatggacgaa aaaaccaccg gttggcgtgg
     1981 tggtcacgtt gttgaaggtc tggctggtga actggaacag ctgcgtgctc gtctggaaca
     2041 ccacccgcag ggtcagcgtg aaccgggatc catgcccgag gatgaccagc gggccacgcg
     2101 caacctcttc attggtaacc tggaccacag cgtatctgag gtggagctgc gaagggcctt
     2161 cgagaaatat ggcatcatcg aggaggtggt catcaagagg cctgcccgtg gccagggcgg
     2221 tgcctatgcc ttcctcaagt tccagaacct ggacatggcc catagggcta aggtggccat
     2281 gtcgggccga gtgattggtc gcaaccccat taagataggc tatggcaagg ccaaccccac
     2341 cactcgtctc tgggtgggtg gcctgggacc taacacgtca ctggcggctc tggcccgaga
     2401 gtttgaccgc tttgggagca ttcggaccat tgatcacgtc aaaggagata gctttgccta
     2461 tattcagtac gagagcttgg acgcagccca ggccgcctgt gctaaaatga ggggttttcc
     2521 cttgggtgga ccagaccgca ggctccgcgt ggattttgcc aaagcagagg agactcggta
     2581 cccccagcag taccagccct cgccactccc tattgcggcc gcatcgtgac tgactgacga
     2641 tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2701 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2761 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     2821 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2881 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2941 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3001 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3061 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3121 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3181 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3241 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     3301 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3361 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3421 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3481 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3541 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     3601 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3661 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3721 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3781 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3841 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3901 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3961 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4021 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4081 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4141 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4201 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4261 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4321 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4381 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4441 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4501 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4561 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4621 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4681 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4741 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4801 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4861 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg cgcaaaacct
     4921 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     4981 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     5041 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     5101 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     5161 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     5221 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5281 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5341 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     5401 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     5461 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     5521 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     5581 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaa
//
