LOCUS       sequence-271566-        4275 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses SFPQ 369-598 with TEV-cleavable hexahistidine tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4275)
  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 4275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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(369-598)
FEATURES             Location/Qualifiers
     source          1..4275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..56
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(94..112)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      108..155
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(326..1117)
                     /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(1118..1209)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1257..1995)
                     /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(2205..3287)
                     /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     3249..3268
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3288..3365)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3411..3430
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3411..3429
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3430..3454
                     /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             3504..3521
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3552..3572
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
       61 ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc
      121 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
      181 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
      241 ctatttaacg accctgccct gaaccgacga ccgggtcatc gtggccggat cttgcggccc
      301 ctcggcttga acgaattgtt agacattatt tgccgactac cttggtgatc tcgcctttca
      361 cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc
      421 caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
      481 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
      541 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
      601 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
      661 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
      721 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
      781 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
      841 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
      901 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
      961 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     1021 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     1081 gttgagtcga tacttcggcg atcaccgctt ccctcatact cttccttttt caatattatt
     1141 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1201 ataaacaaat agctagctca ctcggtcgct acgctccggg cgtgagactg cggcgggcgc
     1261 tgcggacaca tacaaagtta cccacagatt ccgtggataa gcaggggact aacatgtgag
     1321 gcaaaacagc agggccgcgc cggtggcgtt tttccatagg ctccgccctc ctgccagagt
     1381 tcacataaac agacgctttt ccggtgcatc tgtgggagcc gtgaggctca accatgaatc
     1441 tgacagtacg ggcgaaaccc gacaggactt aaagatcccc accgtttccg gcgggtcgct
     1501 ccctcttgcg ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc cgcctttctc
     1561 ccttacggga agtgtggcgc tttctcatag ctcacacact ggtatctcgg ctcggtgtag
     1621 gtcgttcgct ccaagctggg ctgtaagcaa gaactccccg ttcagcccga ctgctgcgcc
     1681 ttatccggta actgttcact tgagtccaac ccggaaaagc acggtaaaac gccactggca
     1741 gcagccattg gtaactggga gttcgcagag gatttgttta gctaaacacg cggttgctct
     1801 tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga tttggttgct gtgctctgcg
     1861 aaagccagtt accacggtta agcagttccc caactgactt aaccttcgat caaaccacct
     1921 ccccaggtgg ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa aggatctcaa
     1981 gaagatcctt tgatcttttc tactgaaccg ctctagattt cagtgcaatt tatctcttca
     2041 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc
     2101 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     2161 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     2221 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2281 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     2341 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     2401 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     2461 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
     2521 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     2581 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     2641 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     2701 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     2761 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     2821 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     2881 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     2941 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     3001 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     3061 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     3121 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     3181 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     3241 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     3301 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     3361 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact
     3421 cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt tgtttaactt
     3481 taataaggag atataccatg aaacatcacc atcaccatca ccccatgagc gattacgaca
     3541 tccccactac tgagaatctt tattttcagg gcgccatggc gcatgctgct gccctttctg
     3601 ttcgtaatct ttcaccttat gtttccaatg aactgttgga agaagccttt agccaatttg
     3661 gtcctattga aagggctgtt gtaatagtgg atgatcgtgg aagatctaca gggaaaggca
     3721 ttgttgaatt tgcttctaag ccagcagcaa gaaaggcatt tgaacgatgc agtgaaggtg
     3781 ttttcttact gacgacaact cctcgtccag tcattgtgga accacttgaa caactagatg
     3841 atgaagatgg tcttcctgaa aaacttgccc agaagaatcc aatgtatcaa aaggagagag
     3901 aaacccctcc tcgttttgcc cagcatggca cgtttgagta cgaatattct cagcgatgga
     3961 agtctttgga tgaaatggaa aaacagcaaa gggaacaagt tgaaaaaaac atgaaagatg
     4021 caaaagacaa attggaaagt gaaatggaag atgcctatca tgaacatcag gcaaatcttt
     4081 tgcgccaaga tctgatgaga cgacaggaag aattaagacg catggaagaa cttcacaatc
     4141 aagaaatgca gaaacgtaaa gaaatgcaat tgaggcaaga ggaggaacga cgtagaagag
     4201 aggaagagat gatgattcgt caacgtgaga tggaagaaca aatgaggcgc caaagagagg
     4261 aaagttacag ctaac
//
