LOCUS       sequence-273993-        5477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of HNRNPCL1 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5477)
  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 5477)
  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: HNRNPCL1_RRM_pGEX
FEATURES             Location/Qualifiers
     source          1..5477
                     /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(1309..1331)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1476..1494)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1562..1666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1667..1735,1736..2527)
                     /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(1885..1904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2698..3286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3187..3206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3440..3457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3530..3607
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3608..4690
                     /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(3627..3646)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4739..4744,4745..4762,4763..4769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4793
                     /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     4801..4817
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4801..4817
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4813..5340
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4833..4850)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4833..4849)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4842..4864)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(5442..5447,5448..5463,5464..5470)
                     /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 ccatggccag caacgttacc aacaagatgg atcctcactc catgaactcc
      901 cgtgtgttca ttgggaatct caacactctt gttgtcaaga aatcggatgt ggaggcgatc
      961 ttttccaagt atggcaaaat tgcgggctgc tctgttcata agggctttgc cttcgttcaa
     1021 tatgataagg agaaaaatgc ccgggctgct gtagcaggag aggatggcag aatgattgct
     1081 agccaggttg tagatattaa cctggctgcg gagccaaaag tgaaccgagg aaacgcaggt
     1141 gtgaaacgat cagcagcgga gatgtacggc tcctcttttg acttggacta tggctttcaa
     1201 cgggattatt atgatggaat gtacagtttc ccagcacgtg taccttaggc ggccgcatcg
     1261 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1321 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1381 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     1441 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     1501 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1561 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1621 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1681 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1741 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1801 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1861 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1921 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1981 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2041 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2101 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2161 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     2221 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2281 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2341 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2401 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2461 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2521 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2581 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2641 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2701 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2761 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2821 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2881 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2941 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3001 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3061 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3361 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3421 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     3481 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     3541 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     3601 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     3661 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     3721 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     3781 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     3841 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     3901 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     3961 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     4021 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     4081 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     4141 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     4201 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     4261 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     4321 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     4381 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     4441 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     4501 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     4561 gttgcccgtc tcactggtga aaagaaaaac cacgctagcg cccaatacgc aaaccgcctc
     4621 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4681 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
     4741 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4801 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     4861 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4921 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4981 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     5041 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     5101 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
     5161 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     5221 gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc
     5281 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     5341 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     5401 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
     5461 tcgtataatg tgtggaa
//
