LOCUS       sequence-271583-        4737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SFPQ 214-598 quadruple mutant (L535A, L539A, L546A, M549A)
            with TEV-cleavable hexahistidine tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4737)
  AUTHORS   Lee M, Sadowska A, Bekere I, Ho D, Gully BS, Lu Y, Iyer KS, 
            Trewhella J, Fox AH, Bond CS
  TITLE     The structure of human SFPQ reveals a coiled-coil mediated polymer 
            essential for functional aggregation in gene regulation.
  JOURNAL   Nucleic Acids Res. 2015 Apr 20;43(7):3826-40. doi: 
            10.1093/nar/gkv156. Epub 2015 Mar 12.
  PUBMED    25765647
REFERENCE   2  (bases 1 to 4737)
  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: 
            "10.1093/nar/gkv156"; journalName: "Nucleic Acids Res"; date: 
            "2015-04-20- 20"; volume: "43"; issue: "7"; pages: "3826-40"
COMMENT     Sequence Label: pCDF11-SFPQ(214-598)_quadruple_mutant
FEATURES             Location/Qualifiers
     source          1..4737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             16..60
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(98..116)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      112..159
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(330..1121)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1122..1213)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1261..1999)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(2209..3291)
                     /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     3253..3272
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3292..3369)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3415..3434
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3415..3433
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3434..3458
                     /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             3508..3525
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3556..3576
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 taactcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg
       61 tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa
      121 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag
      181 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
      241 gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc ggatcttgcg
      301 gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct
      361 ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct
      421 tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc
      481 tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac
      541 caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag
      601 ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca
      661 aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc
      721 aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg
      781 cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg
      841 tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc
      901 gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc
      961 accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg
     1021 tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct
     1081 gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcaatat
     1141 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1201 aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag actgcggcgg
     1261 gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg
     1321 tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca
     1381 gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg
     1441 aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt
     1501 cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt
     1561 tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt
     1621 gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg
     1681 cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact
     1741 ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg
     1801 ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc
     1861 tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc
     1921 acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc
     1981 tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc aatttatctc
     2041 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc
     2101 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     2161 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     2221 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2281 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     2341 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     2401 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     2461 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     2521 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     2581 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     2641 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     2701 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     2761 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     2821 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     2881 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     2941 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     3001 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     3061 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     3121 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     3181 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     3241 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     3301 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     3361 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac
     3421 gactcactat aggggaattg tgagcggata acaattcccc tgtagaaata attttgttta
     3481 actttaataa ggagatatac catgaaacat caccatcacc atcaccccat gagcgattac
     3541 gacatcccca ctactgagaa tctttatttt cagggcgcca tgggcgggcc gaagccaggt
     3601 ggcggcccgg gcctaagtac gcctggcggc caccccaagc cgccgcatcg aggcggcggg
     3661 gagccccgcg ggggccgcca gcaccacccg ccctaccacc agcagcatca ccaggggccc
     3721 ccgcccggcg ggcccggcgg ccgcagcgag gagaagatct cggactcgga ggggtttaaa
     3781 gccaatttgt ctctcttgag gaggcctgga gagaaaactt acacacagcg atgtcggttg
     3841 tttgttggga atctacctgc tgatatcacg gaggatgagt tcaaaagact atttgctaaa
     3901 tatggagaac caggagaagt ttttatcaac aaaggcaaag gattcggatt tattaagctt
     3961 gaatctagag ctttggctga aattgccaaa gccgaactgg atgatacacc catgagaggt
     4021 agacagcttc gagttcgctt tgccacacat gctgctgccc tttctgttcg taatctttca
     4081 ccttatgttt ccaatgaact gttggaagaa gcctttagcc aatttggtcc tattgaaagg
     4141 gctgttgtaa tagtggatga tcgtggaaga tctacaggga aaggcattgt tgaatttgct
     4201 tctaagccag cagcaagaaa ggcatttgaa cgatgcagtg aaggtgtttt cttactgacg
     4261 acaactcctc gtccagtcat tgtggaacca cttgaacaac tagatgatga agatggtctt
     4321 cctgaaaaac ttgcccagaa gaatccaatg tatcaaaagg agagagaaac ccctcctcgt
     4381 tttgcccagc atggcacgtt tgagtacgaa tattctcagc gatggaagtc tttggatgaa
     4441 atggaaaaac agcaaaggga acaagttgaa aaaaacatga aagatgcaaa agacaaattg
     4501 gaaagtgaaa tggaagatgc ctatcatgaa catcaggcaa atcttgcgcg ccaagatgcg
     4561 atgagacgac aggaagaagc aagacgcgcg gaagaacttc acaatcaaga aatgcagaaa
     4621 cgtaaagaaa tgcaattgag gcaagaggag gaacgacgta gaagagagga agagatgatg
     4681 attcgtcaac gtgagatgga agaacaaatg aggcgccaaa gagaggaaag ttacagc
//
