LOCUS       sequence-390706-        4726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His-tagged Drosophila IntS11 amino acids 300-597.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4726)
  AUTHORS   Fujiwara R, Zhai SN, Liang D, Shah AP, Tracey M, Ma XK, Fields CJ, 
            Mendoza-Figueroa MS, Meline MC, Tatomer DC, Yang L, Wilusz JE
  TITLE     IntS6 and the Integrator phosphatase module tune the efficiency of 
            select premature transcription termination events.
  JOURNAL   Mol Cell. 2023 Nov 14:S1097-2765(23)00902-4. doi: 
            10.1016/j.molcel.2023.10.035.
  PUBMED    37995689
REFERENCE   2  (bases 1 to 4726)
  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: "Mol Cell. 
            2023 Nov 14:S1097-2765(23)00902-4. doi: 
            10.1016/j.molcel.2023.10.035."
COMMENT     Sequence Label: pRSFDuet-1 IntS11 AA 300-597
FEATURES             Location/Qualifiers
     source          1..4726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(193..1275)
                     /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     1237..1256
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1276..1353)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1399..1417
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1418..1442
                     /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             1498..1515
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2526..2544
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2545..2569
                     /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             2678..2722
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2760..2778)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2774..2821
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3051..3866)
                     /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(3051..3866)
                     /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     3774..3793
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3867..3958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3974..4723)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3984..4001)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4224..4243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
       61 taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg
      121 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
      181 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      241 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
      301 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
      361 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
      421 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
      481 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
      541 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
      601 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
      661 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
      721 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
      781 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
      841 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
      901 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
      961 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     1021 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     1081 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     1141 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     1201 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     1261 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     1321 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     1381 actcctgcat taggaaatta atacgactca ctatagggga attgtgagcg gataacaatt
     1441 cccctgtaga aataattttg tttaacttta ataaggagat ataccatggg cagcagccat
     1501 caccatcatc accacagcca ggatccgaat tcgagctcgg cgcgcctgca ggtcgacttc
     1561 gacttcaagc acatcaagcc ctttgacaag gcctacatcg acaatcctgg cgcgatggta
     1621 gtgttcgcca cgccaggcat gctgcacgca ggtctttccc tgcagatctt caagaagtgg
     1681 gcgccgaatg agaacaacat ggtgattatg cccggctact gtgtgcaggg aaccgtgggc
     1741 aacaagattc tcggtggcgc caagaaggtg gagttcgaga accgccaggt ggtcgaggtc
     1801 aaaatggccg tggagtacat gagcttctcg gcccatgcag atgccaaagg cattatgcag
     1861 ctaattcaga actgtgagcc aaaaaatgtc atgctcgtcc atggtgaagc ggggaagatg
     1921 aagttcctgc gctcgaagat caaagacgaa ttcaacctgg aaacctacat gccggccaac
     1981 ggcgaaacct gtgtgatttc tacgcctgtg aagataccag tagatgcatc cgtttcactg
     2041 cttaaagcag aggcccgctc ctacaacgcc cagccaccgg atcccaagag gcggcgacta
     2101 atccatggcg tcctcgtaat gaaggacaat cgaataatgc tgcaaaacct aaccgatgcc
     2161 ctaaaggaaa tcggaattaa tcgacatgtt atgcgattta cgtccaaggt caaaatggac
     2221 gactccggac cggtcattcg cacaagcgaa agactgaaga ctctgctcga agaaaagcta
     2281 gccggctgga cggtgacgat gcaggagaac ggctccatag ccatcgagtc cgttgaggtg
     2341 aaggtggagg aggacgagaa ggatcccaag cagaagaaca ttttgatatc gtggaccaac
     2401 caggacgagg acattggagc ctacataccg aatgtgctgc agaatatgtg ctagaagctt
     2461 gcggccgcat aatgcttaag tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc
     2521 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc catcttagta
     2581 tattagttaa gtataagaag gagatataca tatggcagat ctcaattgga tatcggccgg
     2641 ccacgcgatc gctgacgtcg gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt
     2701 tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac
     2761 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2821 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
     2881 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgacaag
     2941 ctgacgaccg ggtctccgca agtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     3061 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     3121 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     3181 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     3241 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     3301 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     3361 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     3421 agacgaaata cgcggtcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     3481 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     3541 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     3601 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     3661 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     3721 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     3781 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     3841 gacgtttccc gttgaatatg gctcatactc ttcctttttc aatattattg aagcatttat
     3901 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3961 ggcatgcagc gctcttccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     4021 cggcgagcgg tgtcagctca ctcaaaagcg gtaatacggt tatccacaga atcaggggat
     4081 aaagccggaa agaacatgtg agcaaaaagc aaagcaccgg aagaagccaa cgccgcaggc
     4141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagccagag
     4201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4321 aagcgtggcg ctttctcata gctcacgctg ttggtatctc agttcggtgt aggtcgttcg
     4381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccat
     4501 tggtaactga tttagaggac tttgtcttga agttatgcac ctgttaaggc taaactgaaa
     4561 gaacagattt tggtgagtgc ggtcctccaa cccacttacc ttggttcaaa gagttggtag
     4621 ctcagcgaac cttgagaaaa ccaccgttgg tagcggtggt ttttctttat ttatgagatg
     4681 atgaatcaat cggtctatca agtcaacgaa cagctattcc gttact
//
