LOCUS       sequence-273977-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of RBM45 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  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: RBM45_Full-length_pGEX
FEATURES             Location/Qualifiers
     source          1..6506
                     /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(2338..2360)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2505..2523)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2591..2695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2696..2764,2765..3556)
                     /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(2914..2933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3727..4315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4216..4235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4469..4486
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4559..4636
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4637..5719
                     /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(4656..4675)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5768..5773,5774..5791,5792..5798)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5806..5822
                     /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     5830..5846
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5830..5846
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5842..6369
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5862..5879)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5862..5878)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5871..5893)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6471..6476,6477..6492,6493..6499)
                     /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 ccatggacga agctggcagc tctgcgagcg gcgggggctt ccgcccgggc
      901 gtggacagcc tggacgaacc gcccaacagc cgcatcttcc ttgtgatcag caagtacaca
      961 cctgagtcgg tgctgaggga gcgcttctcg ccttttggcg acatccagga catctgggtg
     1021 gtgcgggaca agcacaccaa ggagtccaag ggcattgctt tcgtcaagtt cgcccgcagc
     1081 tcacaggcct gcagggccat ggaggagatg catggccagt gcctcggccc caacgacacc
     1141 aagcccatca aggttttcat tgctcagtcc cgatcatctg gaagtcaccg agatgttgaa
     1201 gatgaagaac ttacaagaat ctttgttatg ataccaaagt cctacacaga agaagatctg
     1261 cgggaaaaat ttaaggtgta tggagatatc gagtattgca gcattattaa gaataaagtg
     1321 actggagaaa gtaaaggttt gggctacgta cgatacttaa aaccatcaca agctgcccaa
     1381 gcaatagaaa actgtgatcg aagttttaga gcaatcttgg ctgaacctaa aaataaagca
     1441 tctgaatcct ctgaacaaga ttattatagt aatatgaggc aagaagcttt gggacatgaa
     1501 cctagagtaa atatgtttcc atttgaacaa caatctgaat tttcaagttt tgacaagaat
     1561 gatagccgag gccaggaagc aatctccaaa cgcttgtcag ttgtatcaag agttcctttc
     1621 actgaagaac agcttttcag catttttgat atagtaccag gattggaata ttgtgaagtt
     1681 caacgagatc cttattcaaa ttatggtcat ggagtggttc agtattttaa tgtagcatca
     1741 gctatttatg caaaatacaa attacatgga tttcagtacc ctcctgggaa ccgaataggt
     1801 gtttccttca ttgatgatgg aagtaatgca acagatctcc ttagaaaaat ggcaacacag
     1861 atggtagctg cacagcttgc atcaatggtg tggaataacc caagtcagca acaatttatg
     1921 caatttggag gaagctctgg atcacagttg cctcaaatcc agacagatgt tgtacttcca
     1981 tcatgcaaaa aaaaagctcc tgctgaaact cctgtgaaag aaagactttt tattgtgttt
     2041 aatcctcatc ctttaccttt agacgtatta gaagatatat tctgtcgttt tggtaacctg
     2101 atcgaagttt accttgtgtc aggaaaaaat gtggggtatg ccaagtatgc cgatagaata
     2161 agtgctaatg atgccattgc cactctacat ggaaagattc tgaatggggt gagacttaaa
     2221 gttatgctgg cagattcgcc aagagaagaa tctaacaaac ggcaaagaac ttactgagcg
     2281 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     2341 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2401 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2461 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
     2521 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2581 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2641 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2701 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2761 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2821 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2881 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     2941 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3001 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3061 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3121 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3181 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     3241 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3301 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3361 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3421 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3481 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3541 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3601 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3661 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3721 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3781 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3841 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3901 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3961 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4021 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4081 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4141 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4201 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4261 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4321 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4381 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4441 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4501 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
     4561 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     4621 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     4681 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     4741 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     4801 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     4861 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     4921 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     4981 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     5041 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     5101 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     5161 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     5221 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     5281 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     5341 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     5401 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     5461 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     5521 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     5581 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc acgctagcgc ccaatacgca
     5641 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5701 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     5761 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5821 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
     5881 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     5941 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     6001 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
     6061 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
     6121 cacggttacg atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg
     6181 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
     6241 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaattag cttatcgact
     6301 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
     6361 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     6421 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     6481 atcatcggct cgtataatgt gtggaa
//
