LOCUS       sequence-390002-        7456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged zebrafish IntS6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7456)
  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 7456)
  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: pUb FLAG-Zebrafish IntS6
FEATURES             Location/Qualifiers
     source          1..7456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    27..48
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(63..68,69..86,87..93)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    101..117
                     /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)."
     primer_bind     106..128
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     125..141
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     125..141
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2202..2267
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4877..4918
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4928..4945
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4963..5092
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(5206..5225)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5325..5347
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5385..5403)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5471..5575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5576..5644,5645..6436)
                     /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     complement(5794..5813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6607..7195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7096..7115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7349..7366
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
       61 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      121 cacacaggaa acagctatga ccatgattac gccaagcttt gtcgccggaa cgcagcgaca
      181 gagattccaa tgtgtccgta tctttcaggc ttttgccctt cagttccaga cgaagcgact
      241 ggcgattcgc gtgtggggtc tgcttcaggg tcttgtgaat tagggcgcgc agatcgccga
      301 tgggcgtggc gccggagggc accttcacct tgccgtacgg cttgctgttc ttcgcgttca
      361 aaatctccag ctccattttg ctttcggtgc gcttgcaatc agtactgtcc aaaatcgaaa
      421 atcgccgaac cgtagtgtga ccgtgcgggg ctctgcgaaa ataaactttt ttaggtatat
      481 ggccacacac ggggaaagca cagtggatta tatgttttaa tattataata tgcaggtttt
      541 cattacttat ccagatgtaa gcccacttaa agcgatttaa caattatttg ccgaaagagt
      601 ataaacaaat ttcacttaaa aatggattaa gaaaagctag cttgtgtaag attatgcgca
      661 gcgttgccag atagctccat ttaaaacact tcaaaaacaa taagttttga aaatatatac
      721 ataaatagca gtcgttgccg caacgctcaa cacatcacac ttttaaaaca ccctttacct
      781 acacagaatt actttttaaa tttccagtca agctgcgagt ttcaaaatta tagccggtag
      841 agaagacagt gctatttcaa aagcaaacta acaagggtct taaattccaa aacaccaatc
      901 ctaacaagcc ttggactttt gtaagtttag atcaaaggtg gcattgcatt caatgtcatg
      961 gtaagaagta ggtcgtctag gtagaaatcc tcattcagcc ggtcaagtca gtacgagaaa
     1021 ggtctcaatt tgaaattgtc ttaaaaatat tttattgttt tgtactgtgg tgagtttaaa
     1081 cgaaaaacac aaaaaaaaag tgatacacag aaatcataaa aaattttaat acaaggtatt
     1141 cgtacgtatc aaaaacattt cggcacaatt ttttttctct gtactaaagt gttacgaaca
     1201 ctacggtatt ttttagtgat tttcaacgga caccgaaggt atataaacag cgttcgcgaa
     1261 cggtcgcctt caaaaccaat tgacatttgc agcagcaagt acaagtagaa agtaaagcgc
     1321 aatcagcgaa aaatttatac ttaattgttg gtgattaaag tacaattaaa agaacattct
     1381 cgaaagtcac aagaaacgta agtttttaac tcgctgttac caattagtaa taagagcaac
     1441 aagacgttga gtaatttcaa gaaaaactgc atttcaaggt ctttgttcgg ccattttttt
     1501 ttattcaacg ctctacgtaa ttacaaaata agaaattggc agccacgcat cttgttttcc
     1561 caatgaattg gcatcaaaac gcaaacaaat ctataaataa aacttgcgtg ttgattttcg
     1621 ccaagattta ttggcaaatt gtgaaattcg cagtgacgca tttgaaaatt cgagaaatca
     1681 cgaacgcact cgatcgagca tttgtgtgca tgttattagt tagttgttta gttaattgaa
     1741 gtattttacc aacgaaatcc acttattttt agctgaaata gagtaggttg cttaaacaaa
     1801 gccacgtctg aaaatttctt attgcttgta gttgtgacgt caccatatac acacaaaata
     1861 atgtgtatgc atgcatttca gctgtgtata tatacatgca cacactcgca acacgaaaac
     1921 gatgacgaag caacggaaca aaggtttctc aactaccctt tgttccctgt ttcttcgctt
     1981 tcctttgttc caatattcgt agagggttaa taggggtttc tcaacaaagt tggcgtcgat
     2041 aaataagttt cccattttta ttccccagcc aggaagttag tttcaatagt tttgtaattt
     2101 caacgaaact catttgattt cgtactaatt ttccacatct ctattttctg cccgcagaat
     2161 aatccaaact gcaggtcgac tctagctaga ggaagcttat ggactacaaa gaccatgacg
     2221 gtgattataa agatcatgat atcgattaca aggatgacga tgacaaggat ccactagtcc
     2281 ccgtgctact ttttctaata gacacgtccg cctccatgaa ccagcgctcc catctgggca
     2341 ctagctatct ggacatagcg aagggcgcag ttgagacttt cctgaagctg agaagcagag
     2401 atccggccag caggggagac agatacatgt tagtgagttt cgaggaagcg ccggctggca
     2461 ttaaggctgg atggaaggac agtcatgcca cttttatgac tgagctaagg aacctgcaag
     2521 cagttggact gacatcgttt ggccaagctt taaggacagc ttttgatttg ctcaatctta
     2581 atagattagt ttcagggata gacaactatg gacagggtag aaaccctttt ttcttggaac
     2641 ctgctatcat tgtggccata actgatggca gcaagctgac tggcagctct ggggtacaag
     2701 atgagttaca tttaccgtta acaacaccat tacctggcag tgaactgacc aaagagccct
     2761 ttcgatggga ccagcgactc ttctctttag tgctgcgtgt tccaggccat gcctctgctg
     2821 accctgaacc agtgggtgga gttccactcg actcctcccc tattacaccc atgtgtgagg
     2881 tcaccggagg ccgatcatac agcgtgtttt cccagagaat gctgaaccag tgcctggagt
     2941 ctttggtaca gaagatacag agtggtgtgg tcatcaattt tgagaagact ggacctgatc
     3001 cttctcctac tgaagatggt ccaatagagg tgaagcatgg acctcaagta tggcacagct
     3061 gtcataaatt gatctatgtg agacccaacc ctaaaactgg tgtacctgtt ggccattggc
     3121 ctatccctga atccttctgg cctgaccaga actctcctac tttgcctcca cgtgcagctc
     3181 accctcatgt gaagttttca tgtgtggatt cggagcccat ggtggtagat aaagtgccct
     3241 tcgacaagta tgagctggag ccttccccgc tcacccagta cattctagag aggaaatcac
     3301 cacatacctg ctggcaggtt tttgtgtgta acagtgctaa atatggagaa ctgggtcaac
     3361 cgtttggata tctgaaggct agcacagctc tcaactgtgt caacctgttt gttatgccct
     3421 ataactatcc tgtggtgctt ccattgcttg atgacttaat tgttgtgcat aagttcaagc
     3481 ctccagtgaa gtggcgacag tcttttgaaa attatcttaa aacaatgccc ccctactaca
     3541 tcccagccct gcggaaagca ttaagaatga tgggggctcc aaacctgtta gcagacaaca
     3601 tggaatatgg cctcagctac agtgttgtgt cttatcttaa aaagctcagt cagcaggcga
     3661 agatcgaggc agaccgagtt tgtgcttcag tgggaaaaaa agtggcacca gaaggtggaa
     3721 ttaaacttcg ttgtaggagc tctgctctat ccctggcaaa caggaaggat ttcacacagc
     3781 ttctgcagag catcatggga gatggaccag cacttccaat ggaggcaaac accaaggagt
     3841 ttgctggctt tcagctggct tccttgaata aagcattaaa accacagggc ttgcgcaacc
     3901 cttacgatat cccaagagct catcttctgg accagctgag ccgcatgagg aggaaccttc
     3961 tgcatgccag tatctgtatt ctcaaagggc aagatcaaga tcagcttcac agtgtgccta
     4021 tagctcagat gggcaactat caggacttct tgaagcattg cccatcacct ctgagagagg
     4081 cagaccctga tcagcccaaa cgtctgcaca cctttggtaa cccctttaaa ctggacaaga
     4141 aggccatgat ggtggatgag gcagatgagt ttgttactgg cactcagggc aaaggcaaac
     4201 gtcctggtga gtccagcagt cctactgtgg gaggggcacc taaacgcagg cgctgcatgt
     4261 ctccattgct tcggcctgga cgggcatata cacctcctag aacaccatcc agaactcccg
     4321 ataacaatca cttggaattg gaaaaccata tcaccaatca tctcgagcca aactgcgact
     4381 cggacagcga tataaatcct gtgccggagt ctgaagtgat ccagcaaagc aaccatctcc
     4441 agacagaaga ggttgagaac cagcaggact gcgtgcagga gaatggccag tccagtgaca
     4501 gtgagctgtc tttgggaagc gagggggaag aggaggcccc gcaccgctac cagtacccct
     4561 gccagctgaa gaagatcaga aaccaggaga gccaggagct caactctgaa ctccgagtgc
     4621 tcatcaccaa ggagatcagg aaacctggaa gacgttatga gaaaatcttc taccttctca
     4681 agcaaatcca gggcagtttg gaaacgcacc tgatcttctt gcagagcata atcaaagagg
     4741 ctgcaagatt taagaagagg gtcttaatcg agcagctgga gattttcata gaggagattc
     4801 acttaagagc caacggcatg aatcatctgg acagtttctg agcggccgct cgagtctaga
     4861 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     4921 taccggtcat catcaccatc accattgagt ttatctgact aaatcttagt ttgtattgtc
     4981 atgttttaat acaatatgtt atgtttaaat atgtttttaa taaattttat aaaataattt
     5041 caacttttat tgtaacaaca ttgtccattt acacactcct ttcaagcgcg tgggatcgat
     5101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5161 atgtgagcca tatgggccca tgtgagccat atggtgcact ctcagtacaa tctgctctga
     5221 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5281 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5341 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5401 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5461 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5521 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5581 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5641 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5701 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5761 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5821 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5881 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5941 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6001 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6061 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6121 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6181 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6241 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6301 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6361 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6421 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6481 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6541 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6601 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6661 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6721 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6781 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6841 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6901 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6961 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7021 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7081 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7141 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7201 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7261 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7321 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7381 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7441 caggtttccc gactgg
//
