LOCUS       sequence-273933-        6605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of  NOVA1 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6605)
  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 6605)
  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: NOVA1_Full-length_pGEX
FEATURES             Location/Qualifiers
     source          1..6605
                     /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(2437..2459)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2604..2622)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2690..2794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2795..2863,2864..3655)
                     /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(3013..3032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3826..4414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4315..4334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4568..4585
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4658..4735
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4736..5818
                     /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(4755..4774)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5867..5872,5873..5890,5891..5897)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5905..5921
                     /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     5929..5945
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5929..5945
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5941..6468
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5961..5977)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5970..5992)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6570..6575,6576..6591,6592..6598)
                     /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 ccatgatggc ggcagctccc atccagcaga acgggaccca cactggggtt
      901 cccatagacc tggacccgcc ggactcgcgg aaaaggccgc tggaagcccc ccctgaagcc
      961 ggcagcacca agaggaccaa tacgggcgaa gacggccagt attttctaaa ggttctcata
     1021 cctagttatg ctgctggatc tataattggg aagggaggac agacaattgt tcagttgcaa
     1081 aaagaaactg gagccaccat caagctgtct aagtccaaag atttttaccc aggtactact
     1141 gagcgagtgt gcttgatcca gggaacggtt gaagcactga atgcagttca tggattcatt
     1201 gcagaaaaaa ttcgagaaat gccccaaaat gtggccaaga cagaaccagt cagcattcta
     1261 caaccccaga ccaccgttaa tccagatcgc atcaaacaaa cattgccatc ttccccaact
     1321 accaccaagt cctctccatc tgatcccatg accacctcca gagctaatca ggtaaagatt
     1381 atagttccca acagcacagc aggtctgata atagggaagg gaggtgctac tgtgaaggct
     1441 gtaatggagc agtcaggggc ttgggtgcag ctttcccaga aacctgatgg gatcaacttg
     1501 caagagaggg ttgtcactgt gagtggagaa cctgaacaaa accgaaaagc tgttgaactt
     1561 atcatccaga agatacaaga ggatccacaa agtggcagct gtctcaatat cagttatgcc
     1621 aatgtgacag gtccagtggc aaattccaat ccaaccggat ctccttatgc aaacactgct
     1681 gaagtgttac caactgctgc agcagctgca gggctattag gacatgctaa ccttgctggc
     1741 gttgcagcct ttccagcagt tttatctggc ttcacaggca atgacctggt ggccatcacc
     1801 tctgcactta atacattagc cagctatgga tataatctca acactttagg tttaggtctc
     1861 agtcaagcag cagcaacagg ggctttggct gcagcagctg ccagtgccaa cccagcagca
     1921 gcagcagcca atttattggc cacctatgcc agtgaagcct cagccagtgg cagcacagct
     1981 ggtggtacgg cggggacatt tgcattaggt agcctggctg ctgctactgc tgcaaccaat
     2041 ggatattttg gagctgcttc tcccctagct gccagtgcca ttctaggaac agaaaagtcc
     2101 acagatggat ccaaggatgt agttgaaata gcagtgccag aaaacttagt tggtgcaata
     2161 cttggcaaag gagggaaaac attagtggaa taccaggagt tgactggtgc aaggatacag
     2221 atctccaaaa aaggagaatt cgtacctggc acaaggaatc ggaaggtaac cattactgga
     2281 acaccagctg caacacaggc tgctcaatat ttaattacac aaaggatcac atatgagcaa
     2341 ggagttcggg ctgccaatcc tcagaaagtg ggttaagcgg ccgcatcgtg actgactgac
     2401 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2461 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2521 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     2581 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2641 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2701 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2761 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2821 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2881 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2941 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3001 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     3061 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3121 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3181 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3241 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3301 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     3361 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3421 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3481 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3541 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3601 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3661 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3721 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3781 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3841 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3901 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3961 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4021 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4081 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4141 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4201 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4261 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4321 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4381 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4441 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4501 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4561 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4621 tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat ggcgcaaaac
     4681 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     4741 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     4801 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     4861 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     4921 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     4981 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     5041 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     5101 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     5161 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     5221 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     5281 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     5341 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     5401 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     5461 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     5521 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     5581 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     5641 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     5701 actggtgaaa agaaaaacca cgctagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5761 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5821 gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5881 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5941 atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6001 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6061 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     6121 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     6181 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     6241 tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     6301 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     6361 cgaattattt ttgatggcgt tggaattagc ttatcgactg cacggtgcac caatgcttct
     6421 ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa
     6481 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac
     6541 ggttctggca aatattctga aatgagctgt tgacaattaa tcatcggctc gtataatgtg
     6601 tggaa
//
