LOCUS       sequence-273911-        6209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of ILF2 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6209)
  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 6209)
  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: ILF2_DZF_pGEX
FEATURES             Location/Qualifiers
     source          1..6209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(496..501,502..519,520..526)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    534..550
                     /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     558..574
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     558..574
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             570..1097
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(590..607)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(590..606)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(599..621)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1199..1204,1205..1220,1221..1227)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1235..1251
                     /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             1274..1927
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1885..1907
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1934..1957
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1967..2080
                     /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(3275..3297)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3442..3460)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3528..3632
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3633..3701,3702..4493)
                     /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(3851..3870)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4664..5252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5153..5172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5406..5423
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5496..5573
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5574..6209,1..447)
                     /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(5593..5612)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
       61 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
      121 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
      181 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
      241 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
      301 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac gctagcgccc
      361 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      421 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
      481 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      541 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg
      601 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      661 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      721 agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc
      781 cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact
      841 ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca
      901 atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg
      961 atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaattagct
     1021 tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat
     1081 ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg
     1141 gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt
     1201 gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca
     1261 ggaaacagta ttcatgtccc ctatactagg ttattggaaa attaagggcc ttgtgcaacc
     1321 cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt atgagcgcga
     1381 tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc
     1441 ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac gttatatagc
     1501 tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt caatgcttga
     1561 aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta aagactttga
     1621 aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg
     1681 tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact tcatgttgta
     1741 tgacgctctt gatgttgttt tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt
     1801 agtttgtttt aaaaaacgta ttgaagctat cccacaaatt gataagtact tgaaatccag
     1861 caagtatata gcatggcctt tgcagggctg gcaagccacg tttggtggtg gcgaccatcc
     1921 tccaaaatcg gatctggaag ttctgttcca ggggcccctg ggattaatgg acgaaaaaac
     1981 caccggttgg cgtggtggtc acgttgttga aggtctggct ggtgaactgg aacagctgcg
     2041 tgctcgtctg gaacaccacc cgcagggtca gcgtgaaccg ggatccagag gaggcccagg
     2101 aggagggttc aggccctttg taccacatat cccatttgac ttctatttgt gtgaaatggc
     2161 ctttccccgg gtcaagccag cacctgatga aacttccttc agtgaggcct tgctgaagag
     2221 gaatcaggac ctggctccca attctgctga acaggcatct atcctttctc tggtgacaaa
     2281 aataaacaat gtgattgata atctgattgt ggctccaggg acatttgaag tgcaaattga
     2341 agaagttcga caggtgggat cctataaaaa ggggacaatg actacaggac acaatgtggc
     2401 tgacctggtg gtgatactca agattctgcc aacgttggaa gctgttgctg ccctggggaa
     2461 caaagtcgtg gaaagcctaa gagcacagga tccttctgaa gttttaacca tgctgaccaa
     2521 cgaaactggc tttgaaatca gttcttctga tgctacagtg aagattctca ttacaacagt
     2581 gccacccaat cttcgaaaac tggatccaga actccatttg gatatcaaag tattgcagag
     2641 tgccttagca gccatccgac atgcccgctg gttcgaggaa aatgcttctc agtccacagt
     2701 taaagttctc atcagactac tgaaggactt gaggattcgt tttcctggct ttgagcccct
     2761 cacaccctgg atccttgacc tactaggcca ttatgctgtg atgaacaacc ccaccagaca
     2821 gcctttggcc ctaaacgttg catacaggcg ctgcttgcag attctggctg caggactgtt
     2881 cctgccaggt tcagtgggta tcactgaccc ctgtgagagt ggcaacttta gagtacacac
     2941 agtcatgacc ctagaacagc aggacatggt ctgctataca gctcagactc tcgtccgaat
     3001 cctctcacat ggtggcttta ggaagatcct tggccaggag ggtgatgcca gctatcttgc
     3061 ttctgaaata tctacctggg atggagtgat agtaacacct tcagaaaagg cttatgagaa
     3121 gccaccagag aagaaggaag gagaggaaga agaggagaat acagaagaac cacctcaagg
     3181 agaggaagaa gaaagcatgg aaactcagga gtgagcggcc gcatcgtgac tgactgacga
     3241 tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3301 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3361 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     3421 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3481 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3541 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3601 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3661 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3721 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3781 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3841 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     3901 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3961 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4021 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4081 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4141 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     4201 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4261 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4321 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4381 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4441 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4501 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4561 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4621 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4681 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4741 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4801 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4861 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4921 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4981 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5041 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5101 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5161 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5221 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5281 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5341 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5401 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     5461 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg cgcaaaacct
     5521 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     5581 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     5641 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     5701 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     5761 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     5821 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5881 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5941 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     6001 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     6061 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     6121 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     6181 aatatctcac tcgcaatcaa attcagccg
//
