LOCUS       sequence-273978-        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of SNRPA with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  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 5930)
  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: SNRPA_Full-length_pGEX
FEATURES             Location/Qualifiers
     source          1..5930
                     /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(1762..1784)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1929..1947)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2015..2119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2120..2188,2189..2980)
                     /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(2338..2357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3151..3739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3640..3659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3893..3910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3983..4060
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4061..5143
                     /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(4080..4099)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5192..5197,5198..5215,5216..5222)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5230..5246
                     /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     5254..5270
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5254..5270
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5266..5793
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5286..5303)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5286..5302)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5295..5317)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5895..5900,5901..5916,5917..5923)
                     /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 ccatggcagt tcccgagacc cgccctaacc acactattta tatcaacaac
      901 ctcaatgaga agatcaagaa ggatgagcta aaaaagtccc tgtacgccat cttctcccag
      961 tttggccaga tcctggatat cctggtatca cggagcctga agatgagggg ccaggccttt
     1021 gtcatcttca aggaggtcag cagcgccacc aacgccctgc gctccatgca gggtttccct
     1081 ttctatgaca aacctatgcg tatccagtat gccaagaccg actcagatat cattgccaag
     1141 atgaaaggca ccttcgtgga gcgggaccgc aagcgggaga agaggaagcc caagagccag
     1201 gagaccccgg ccaccaagaa ggctgtgcaa ggcgggggag ccacccccgt ggtgggggct
     1261 gtccaggggc ctgtcccggg catgccgccg atgactcagg cgccccgcat tatgcaccac
     1321 atgccgggcc agccgcccta catgccgccc cctggtatga tccccccgcc aggccttgca
     1381 cctggccaga tcccaccagg ggccatgccc ccgcagcagc ttatgccagg acagatgccc
     1441 cctgcccagc ctctttctga gaatccaccg aatcacatct tgttcctcac caacctgcca
     1501 gaggagacca acgagctcat gctgtccatg cttttcaatc agttccctgg cttcaaggag
     1561 gtccgtctgg tacccgggcg gcatgacatc gccttcgtgg agtttgacaa tgaggtacag
     1621 gcaggggcag ctcgcgatgc cctgcagggc tttaagatca cgcagaacaa cgccatgaag
     1681 atctcctttg ccaagaagtg agcggccgca tcgtgactga ctgacgatct gcctcgcgcg
     1741 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     1801 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     1861 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
     1921 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     1981 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2041 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2101 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2161 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2221 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2281 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2341 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     2401 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2461 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2521 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2581 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2641 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2701 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2761 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2821 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2881 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2941 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3001 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3061 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3301 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3781 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3841 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3901 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3961 tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc
     4021 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     4081 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     4141 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     4201 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     4261 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     4321 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     4381 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     4441 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     4501 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     4561 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     4621 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     4681 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     4741 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     4801 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     4861 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     4921 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     4981 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     5041 aaccacgcta gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5101 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5161 taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     5221 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     5281 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5341 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5401 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
     5461 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
     5521 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
     5581 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
     5641 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
     5701 ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca
     5761 tcggaagctg tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag
     5821 gcgcactccc gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat
     5881 tctgaaatga gctgttgaca attaatcatc ggctcgtata atgtgtggaa
//
