LOCUS       sequence-403166-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express sumo-tagged human SRSF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  AUTHORS   Fargason T, De Silva NIU, King E, Zhang Z, Paul T, Shariq J, 
            Zaharias S, Zhang J
  TITLE     Peptides that Mimic RS repeats modulate phase separation of SRSF1, 
            revealing a reliance on combined stacking and electrostatic 
            interactions.
  JOURNAL   Elife. 2023 Mar 2;12:e84412. doi: 10.7554/eLife.84412.
  PUBMED    36862748
REFERENCE   2  (bases 1 to 6357)
  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"; journalName: "Elife."; 
            date: "2023-03-2"; volume: "12:e84412. doi"; pages: " 
            10.7554/eLife.84412"
COMMENT     Sequence Label: pSMT3_SRSF1
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             157..450
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             1222..1239
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1292..1310)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1306..1353
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1390..1845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1477..1496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1687..1708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1937..2752)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1937..2752)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2660..2679
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2744..2763
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2874..3462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3363..3382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3616..3633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3648..3787
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3713..3732)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3873..3895
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3889..4080)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4889..5971)
                     /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     5933..5952
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5972..6049)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6255..6274
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg gctagcatgt cggactcaga agtcaatcaa
      181 gaagctaagc cagaggtcaa gccagaagtc aagcctgaga ctcacatcaa tttaaaggtg
      241 tccgatggat cttcagagat cttcttcaag atcaaaaaga ccactccttt aagaaggctg
      301 atggaagcgt tcgctaaaag acagggtaag gaaatggact ccttaagatt cttgtacgac
      361 ggtattagaa ttcaagctga tcagacccct gaagatttgg acatggagga taacgatatt
      421 attgaggctc acagagaaca gattggtgga tccatgtcgg gaggtggtgt gattcgtggc
      481 cccgcaggga acaacgattg ccgcatctac gtgggtaact tacctccaga catccgaacc
      541 aaggacattg aggacgtgtt ctacaaatac ggcgctatcc gcgacatcga cctcaagaat
      601 cgccgcgggg gaccgccctt cgccttcgtt gagttcgagg acccgcgaga cgcggaagac
      661 gcggtgtatg gtcgcgacgg ctatgattac gatgggtacc gtctgcgggt ggagtttcct
      721 cgaagcggcc gtggaacagg ccgaggcggc ggcgggggtg gaggtggcgg agctccccga
      781 ggtcgctatg gccccccatc caggcggtct gaaaacagag tggttgtctc tggactgcct
      841 ccaagtggaa gttggcagga tttaaaggat cacatgcgtg aagcaggtga tgtatgttat
      901 gctgatgttt accgagatgg cactggtgtc gtggagtttg tacggaaaga agatatgacc
      961 tatgcagttc gaaaactgga taacactaag tttagatctc atgagggaga aactgcctac
     1021 atccgggtta aagttgatgg gccccgtagc ccgagctatg gccgttctcg atctcgaagc
     1081 cgtagtcgta gcagaagccg tagcagaagc aacagcagga gtcgcagtta ctccccaagg
     1141 agaagcagag gatcaccacg ctattctccc cgtcatagca gatctcgctc tcgtacataa
     1201 aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     1261 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     1321 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     1381 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1441 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1501 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1561 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1621 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1681 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1741 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1801 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     1861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1921 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     1981 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     2041 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     2101 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     2161 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     2221 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     2281 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     2341 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     2401 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     2461 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     2521 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     2581 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     2641 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2701 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     2761 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     2821 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2881 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2941 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3001 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3061 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3121 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3181 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3241 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3301 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3361 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3421 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3481 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3541 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3601 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3661 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     3721 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     3781 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3841 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3901 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     3961 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4021 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4081 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     4141 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     4201 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     4261 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     4321 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     4381 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     4441 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     4501 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     4561 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     4621 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     4681 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     4741 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     4801 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     4861 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4921 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4981 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     5041 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     5101 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     5161 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     5221 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     5281 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     5341 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     5401 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     5461 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     5521 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     5581 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     5641 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     5701 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     5761 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     5821 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     5881 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     5941 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     6001 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     6061 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     6121 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     6181 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     6241 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     6301 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaat
//
