LOCUS       sequence-273905-        6152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant expression of PCBP1 with dual-affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6152)
  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 6152)
  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: PCBP1_Full-length_pGEX
FEATURES             Location/Qualifiers
     source          1..6152
                     /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(1984..2006)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2151..2169)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2237..2341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2342..2410,2411..3202)
                     /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(2560..2579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3373..3961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3862..3881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4115..4132
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4205..4282
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4283..5365
                     /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(4302..4321)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5414..5419,5420..5437,5438..5444)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5452..5468
                     /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     5476..5492
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5476..5492
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5488..6015
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5508..5525)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5508..5524)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5517..5539)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6117..6122,6123..6138,6139..6145)
                     /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 ccatggatgc cggtgtgact gaaagtggac taaatgtgac tctcaccatt
      901 cggcttctta tgcacggaaa ggaagtagga agcatcattg ggaagaaagg ggagtcggtt
      961 aagaggatcc gcgaggagag tggcgcgcgg atcaacatct cggaggggaa ttgtccggag
     1021 agaatcatca ctctgaccgg ccccaccaat gccatcttta aggctttcgc tatgatcatc
     1081 gacaagctgg aggaagatat caacagctcc atgaccaaca gtaccgcggc cagcaggccc
     1141 ccggtcaccc tgaggctggt ggtgccggcc acccagtgcg gctccctgat tgggaaaggc
     1201 gggtgtaaga tcaaagagat ccgcgagagt acgggggcgc aggtccaggt ggcgggggat
     1261 atgctgccca actccaccga gcgggccatc accatcgctg gcgtgccgca gtctgtcacc
     1321 gagtgtgtca agcagatttg cctggtcatg ctggagacgc tctcccagtc tccgcaaggg
     1381 agagtcatga ccattccgta ccagcccatg ccggccagct ccccagtcat ctgcgcgggc
     1441 ggccaagatc ggtgcagcga cgctgcgggc tacccccatg ccacccatga cctggaggga
     1501 ccacctctag atgcctactc gattcaagga caacacacca tttctccgct cgatctggcc
     1561 aagctgaacc aggtggcaag acaacagtct cactttgcca tgatgcacgg cgggaccgga
     1621 ttcgccggaa ttgactccag ctctccagag gtgaaaggct attgggcaag tttggatgca
     1681 tctactcaaa ccacccatga actcaccatt ccaaataact taattggctg cataatcggg
     1741 cgccaaggcg ccaacattaa tgagatccgc cagatgtccg gggcccagat caaaattgcc
     1801 aacccagtgg aaggctcctc tggtaggcag gttactatca ctggctctgc tgccagtatt
     1861 agtctggccc agtatctaat caatgccagg ctttcctctg agaagggcat ggggtgcagc
     1921 taggcggccg catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga
     1981 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2041 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     2101 gacccagtca cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat
     2161 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2221 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2281 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2341 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2401 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2461 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2521 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2581 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2641 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2701 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2761 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2821 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2881 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2941 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3001 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3061 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3121 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3181 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3241 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3301 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3361 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3421 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3481 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3541 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3601 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3661 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3721 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3781 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3841 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3901 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3961 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4021 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4081 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4141 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa
     4201 ttccgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     4261 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     4321 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     4381 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     4441 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     4501 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     4561 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     4621 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     4681 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     4741 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     4801 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     4861 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     4921 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     4981 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     5041 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     5101 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     5161 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     5221 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccacgc tagcgcccaa
     5281 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5341 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     5401 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     5461 gataacaatt tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt
     5521 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     5581 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5641 ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg
     5701 gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg
     5761 cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat
     5821 ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat
     5881 gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta
     5941 tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg
     6001 ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga
     6061 taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga
     6121 caattaatca tcggctcgta taatgtgtgg aa
//
