LOCUS       sequence-271590-        6149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NONO 53-312 in the second site; for co-expression with 
            His-tagged protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6149)
  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 6149)
  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: pET-Duet-1-NONO(53-312)
FEATURES             Location/Qualifiers
     source          1..6149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    233..257
                     /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             1098..1142
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1180..1198)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1194..1241
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1278..1733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1365..1384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1575..1596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1848..2639,2640..2708))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2471..2490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2709..2801)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2882..3470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3371..3390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3624..3641
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3656..3795
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3721..3740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3881..3903
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3897..4088)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4660..5742)
                     /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     5704..5723
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5743..5820)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6026..6045
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(6133..6149,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
      301 tggaaggctt gactattgac ctgaagaatt ttagaaaacc aggagagaag accttcaccc
      361 aacgaagccg tctttttgtg ggaaatcttc ctcccgacat cactgaggaa gaaatgagga
      421 aactatttga gaaatatgga aaggcaggcg aagtcttcat tcataaggat aaaggatttg
      481 gctttatccg cttggaaacc cgaaccctag cggagattgc caaagtggag ctggacaata
      541 tgccactccg tggaaagcag ctgcgtgtgc gctttgcctg ccatagtgca tcccttacag
      601 ttcgaaacct tcctcagtat gtgtccaacg aactgctgga agaagccttt tctgtgtttg
      661 gccaggtaga gagggctgta gtcattgtgg atgatcgagg aaggccctca ggaaaaggca
      721 ttgttgagtt ctcagggaag ccagctgctc ggaaagctct ggacagatgc agtgaaggct
      781 ccttcctgct aaccacattt cctcgtcctg tgactgtgga gcccatggac cagttagatg
      841 atgaagaggg acttccagag aagctggtta taaaaaacca gcaatttcac aaggaacgag
      901 agcagccacc cagatttgca cagcctggct cctttgagta tgaatatgcc atgcgctgga
      961 aggcactcat tgagatggag aagcagcagc aggaccaagt ggaccgcaac atcaaggagg
     1021 ctcgtgagaa gctggagatg gagatggaag ctgcacgcca tgagcaccag gtcatgctaa
     1081 tgtaactcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact
     1141 cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa taactagcat
     1201 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     1261 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1321 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1381 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1441 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1501 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1561 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1621 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     1681 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttctggcgg
     1741 cacgatggca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1801 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1861 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1921 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1981 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2041 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2101 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2161 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2221 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2281 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2341 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2401 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2461 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2521 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2581 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2641 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2701 atactcatac tcttcctttt tcaatcatga ttgaagcatt tatcagggtt attgtctcat
     2761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggtcatg accaaaatcc
     2821 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2881 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2941 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3001 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3061 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3121 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3181 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3241 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3301 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3361 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3421 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3481 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3541 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3601 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3661 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3721 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3781 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3841 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3901 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3961 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4021 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4081 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4141 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4201 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4261 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4321 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4381 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4441 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4501 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgctagt catgccccgc
     4561 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     4621 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4681 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4741 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     4801 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     4861 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     4921 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     4981 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5041 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     5101 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     5161 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     5221 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     5281 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     5341 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5401 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     5461 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     5521 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     5581 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     5641 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     5701 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     5761 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     5821 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     5881 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     5941 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6001 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6061 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg
     6121 atcccgcgaa attaatacga ctcactata
//
