LOCUS       sequence-275061-        5978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of RBMS3 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5978)
  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 5978)
  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: RBMS3_2xRRM_pGEX
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(524..529,530..547,548..554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    562..578
                     /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     586..602
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     586..602
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             598..1125
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(618..635)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(618..634)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(627..649)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1227..1232,1233..1248,1249..1255)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1263..1279
                     /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             1302..1955
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1913..1935
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1962..1985
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1995..2108
                     /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(3072..3094)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3239..3257)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3325..3429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3430..3498,3499..4290)
                     /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(3648..3667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4461..5049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4950..4969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5203..5220
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5293..5370
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5371..5978,1..475)
                     /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(5390..5409)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
       61 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
      121 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
      181 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
      241 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
      301 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
      361 ggtgaaaaga aaaaccacgc tagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      421 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      481 acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
      541 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
      601 accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
      661 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
      721 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt
      781 gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag
      841 gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac
      901 accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg
      961 ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga
     1021 attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa tgcttctggc
     1081 gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc
     1141 gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt
     1201 tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg
     1261 aattgtgagc ggataacaat ttcacacagg aaacagtatt catgtcccct atactaggtt
     1321 attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa
     1381 aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg
     1441 aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac
     1501 agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa
     1561 aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt
     1621 cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac
     1681 ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc
     1741 atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc
     1801 caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc
     1861 cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc
     1921 aagccacgtt tggtggtggc gaccatcctc caaaatcgga tctggaagtt ctgttccagg
     1981 ggcccctggg attaatggac gaaaaaacca ccggttggcg tggtggtcac gttgttgaag
     2041 gtctggctgg tgaactggaa cagctgcgtg ctcgtctgga acaccacccg cagggtcagc
     2101 gtgaaccggg atccatgggc aaacgcctgg atcagccaca aatgtacccc cagtacactt
     2161 actactatcc tcattatctc caaaccaagc agtcctatgc accagctccc caccccatgg
     2221 ctcctcccag ccccagcaca aacagcagca gcaacaacag cagcaacaac agcagcgggg
     2281 aacagttgag taaaaccaac ctgtacattc gaggcctccc accaggcacc actgaccagg
     2341 acctaatcaa gctgtgccaa ccgtatggaa aaattgtatc tacaaaggca attcttgaca
     2401 aaaacacaaa tcagtgcaaa ggttatggtt ttgtagattt tgacagtcct gcagccgcac
     2461 agaaagcggt agcatctctc aaggcaaatg gcgtgcaggc acagatggct aagcaacaag
     2521 agcaagaccc aacaaaccta tacatctcaa atctccccat ttctatggat gagcaggagc
     2581 ttgagaatat gctgaaaccc tttggacatg tcatttccac aagaatacta agagacgcta
     2641 atggagtcag cagaggtgtt ggctttgcca gaatggagtc tactgaaaaa tgtgaagtgg
     2701 taattcaaca ttttaatgga aaatatctga aaacaccacc aggcatccca gcccccagtg
     2761 agcctttgct gtgcaaattc gctgatggag gacaaaagaa gcgacagaat caaagcaaat
     2821 atacccagaa tgggaggcct tggcccaggg aaggagaggc tggcatggct ttgacctatg
     2881 accccacagc tgccatacag aatggatttt attcttcacc gtacagtatt gcaaccaacc
     2941 gcatgattcc acagacatct atcacgccat tcattgctgc ttcccctgtc tccacatacc
     3001 aggtccagtg agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat
     3061 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     3121 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     3181 gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc
     3241 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3301 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3361 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3421 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3481 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3541 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3601 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3661 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3721 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3781 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3841 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3901 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3961 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4021 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4081 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4141 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4201 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4261 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4321 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4381 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4441 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4501 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4561 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4621 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4681 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4741 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4801 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4861 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4921 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4981 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5041 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5101 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5161 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5221 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5281 cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
     5341 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     5401 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     5461 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     5521 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     5581 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     5641 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     5701 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     5761 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     5821 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     5881 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     5941 tctgtctcgg cgcgtctgcg tctggctggc tggcataa
//
