LOCUS       sequence-275414-        6503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of RBM6 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6503)
  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 6503)
  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: RBM6_RRM_pGEX
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(157..162,163..180,181..187)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    195..211
                     /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     219..235
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     219..235
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             231..758
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(251..268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(251..267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(260..282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(860..865,866..881,882..888)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    896..912
                     /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             935..1588
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1546..1568
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1595..1618
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1628..1741
                     /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(3230..3252)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3397..3415)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3483..3587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3588..3656,3657..4448)
                     /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(3806..3825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4619..5207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5108..5127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5361..5378
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5451..5528
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5529..6503,1..108)
                     /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(5548..5567)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 agaaaaacca cgctagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
       61 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      121 taatgtaagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      181 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      241 ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      301 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      361 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt
      421 ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata
      481 ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg
      541 tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt
      601 actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt
      661 ttgatggcgt tggaattagc ttatcgactg cacggtgcac caatgcttct ggcgtcaggc
      721 agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc
      781 tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca
      841 aatattctga aatgagctgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg
      901 agcggataac aatttcacac aggaaacagt attcatgtcc cctatactag gttattggaa
      961 aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga
     1021 agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg
     1081 tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat
     1141 ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg
     1201 tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat
     1261 tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat
     1321 gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac
     1381 ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg
     1441 cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat
     1501 tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac
     1561 gtttggtggt ggcgaccatc ctccaaaatc ggatctggaa gttctgttcc aggggcccct
     1621 gggattaatg gacgaaaaaa ccaccggttg gcgtggtggt cacgttgttg aaggtctggc
     1681 tggtgaactg gaacagctgc gtgctcgtct ggaacaccac ccgcagggtc agcgtgaacc
     1741 gggatcccgc ttaggacatc aaaagagaga agcagaaagg tatctgcctc cttctcgaag
     1801 ggaagggcca actttccgaa gagaccgaga gagggagtca tggtctggag agacacgcca
     1861 ggatggagag agcaaaacta tcatgctaaa gcgtatctat cgttccacac cacctgaggt
     1921 gatagtggaa gtgctggagc cctatgtccg ccttactact gccaacgtcc gtatcatcaa
     1981 gaacagaaca ggccctatgg ggcataccta tggctttatt gacctcgact cccatgcgga
     2041 agctcttcgt gtggtgaaga tcttacagaa ccttgatccg ccatttagca ttgatgggaa
     2101 gatggtagct gtaaacctgg ccactggaaa acgaagaaat gattctgggg accattctga
     2161 ccacatgcat tactatcagg gtaaaaaata tttccgagat aggaggggag gtggcagaaa
     2221 ttcagactgg tcttcagata caaatcgaca aggacaacag tcatcatctg actgctacat
     2281 atatgattct gctactggct actattatga ccccttggca ggaacttatt atgaccccaa
     2341 tacccagcaa gaagtctatg tgccccagga tcctggatta cctgaggaag aagagatcaa
     2401 ggaaaaaaaa cccaccagtc aaggaaagtc aagtagcaag aaggaaatgt ctaaaagaga
     2461 tggcaaggag aaaaaagaca gaggagtgac gaggtttcag gaaaatgcca gtgaagggaa
     2521 ggcccctgca gaagacgtct ttaagaagcc cctgcctcct actgtgaaga aggaagagag
     2581 tccccctcca cctaaagtgg taaacccact gatcggcctc ttgggtgaat atggaggaga
     2641 cagtgactat gaggaggaag aagaggagga acagacccct cccccacagc cccgcacagc
     2701 acagccccag aagcgagagg agcaaaccaa gaaggagaat gaagaagaca aactcactga
     2761 ctggaataaa ctggcttgtc tgctttgcag aaggcagttt cccaataaag aagttctgat
     2821 caaacaccag cagctgtcag acctgcacaa gcaaaacctg gaaatccacc ggaagataaa
     2881 acagtctgag caggagctag cctatctgga aaggagagaa cgagagggaa agtttaaagg
     2941 aagaggaaat gatcgcaggg aaaagctcca gtcttttgac tctccagaaa ggaaacggat
     3001 taagtactcc agggaaactg acagtgatcg taaacttgtt gataaagaag atatcgacac
     3061 tagcagcaaa ggaggctgtg tccaacaggc tactggctgg aggaaaggga caggcctggg
     3121 atatggccat cctggattgg cttcatcaga ggaggctgaa ggccggtgag cggccgcatc
     3181 gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3241 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3301 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     3361 gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     3421 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3541 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3601 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3661 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3721 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3781 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     3841 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3901 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3961 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4021 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4081 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     4141 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4201 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4261 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4321 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4381 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4441 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4741 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5281 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5341 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     5401 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg
     5461 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     5521 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     5581 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     5641 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     5701 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     5761 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     5821 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     5881 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     5941 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     6001 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     6061 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     6121 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     6181 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     6241 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     6301 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     6361 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     6421 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     6481 tgttgcccgt ctcactggtg aaa
//
