LOCUS       sequence-390001-        7571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged zebrafish IntS6L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7571)
  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 7571)
  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 IntS6L
FEATURES             Location/Qualifiers
     source          1..7571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    64..85
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(100..105,106..123,124..130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    138..154
                     /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     143..165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     162..178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     162..178
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2239..2304
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5029..5070
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5080..5097
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5115..5244
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(5358..5377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5477..5499
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5537..5555)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5623..5727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5728..5796,5797..6588)
                     /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(5946..5965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6759..7347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7248..7267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7501..7518
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
       61 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      121 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
      181 tgattacgcc aagctttgtc gccggaacgc agcgacagag attccaatgt gtccgtatct
      241 ttcaggcttt tgcccttcag ttccagacga agcgactggc gattcgcgtg tggggtctgc
      301 ttcagggtct tgtgaattag ggcgcgcaga tcgccgatgg gcgtggcgcc ggagggcacc
      361 ttcaccttgc cgtacggctt gctgttcttc gcgttcaaaa tctccagctc cattttgctt
      421 tcggtgcgct tgcaatcagt actgtccaaa atcgaaaatc gccgaaccgt agtgtgaccg
      481 tgcggggctc tgcgaaaata aactttttta ggtatatggc cacacacggg gaaagcacag
      541 tggattatat gttttaatat tataatatgc aggttttcat tacttatcca gatgtaagcc
      601 cacttaaagc gatttaacaa ttatttgccg aaagagtata aacaaatttc acttaaaaat
      661 ggattaagaa aagctagctt gtgtaagatt atgcgcagcg ttgccagata gctccattta
      721 aaacacttca aaaacaataa gttttgaaaa tatatacata aatagcagtc gttgccgcaa
      781 cgctcaacac atcacacttt taaaacaccc tttacctaca cagaattact ttttaaattt
      841 ccagtcaagc tgcgagtttc aaaattatag ccggtagaga agacagtgct atttcaaaag
      901 caaactaaca agggtcttaa attccaaaac accaatccta acaagccttg gacttttgta
      961 agtttagatc aaaggtggca ttgcattcaa tgtcatggta agaagtaggt cgtctaggta
     1021 gaaatcctca ttcagccggt caagtcagta cgagaaaggt ctcaatttga aattgtctta
     1081 aaaatatttt attgttttgt actgtggtga gtttaaacga aaaacacaaa aaaaaagtga
     1141 tacacagaaa tcataaaaaa ttttaataca aggtattcgt acgtatcaaa aacatttcgg
     1201 cacaattttt tttctctgta ctaaagtgtt acgaacacta cggtattttt tagtgatttt
     1261 caacggacac cgaaggtata taaacagcgt tcgcgaacgg tcgccttcaa aaccaattga
     1321 catttgcagc agcaagtaca agtagaaagt aaagcgcaat cagcgaaaaa tttatactta
     1381 attgttggtg attaaagtac aattaaaaga acattctcga aagtcacaag aaacgtaagt
     1441 ttttaactcg ctgttaccaa ttagtaataa gagcaacaag acgttgagta atttcaagaa
     1501 aaactgcatt tcaaggtctt tgttcggcca ttttttttta ttcaacgctc tacgtaatta
     1561 caaaataaga aattggcagc cacgcatctt gttttcccaa tgaattggca tcaaaacgca
     1621 aacaaatcta taaataaaac ttgcgtgttg attttcgcca agatttattg gcaaattgtg
     1681 aaattcgcag tgacgcattt gaaaattcga gaaatcacga acgcactcga tcgagcattt
     1741 gtgtgcatgt tattagttag ttgtttagtt aattgaagta ttttaccaac gaaatccact
     1801 tatttttagc tgaaatagag taggttgctt aaacaaagcc acgtctgaaa atttcttatt
     1861 gcttgtagtt gtgacgtcac catatacaca caaaataatg tgtatgcatg catttcagct
     1921 gtgtatatat acatgcacac actcgcaaca cgaaaacgat gacgaagcaa cggaacaaag
     1981 gtttctcaac taccctttgt tccctgtttc ttcgctttcc tttgttccaa tattcgtaga
     2041 gggttaatag gggtttctca acaaagttgg cgtcgataaa taagtttccc atttttattc
     2101 cccagccagg aagttagttt caatagtttt gtaatttcaa cgaaactcat ttgatttcgt
     2161 actaattttc cacatctcta ttttctgccc gcagaataat ccaaactgca ggtcgactct
     2221 agctagagga agcttatgga ctacaaagac catgacggtg attataaaga tcatgatatc
     2281 gattacaagg atgacgatga caaggatcca ctagtcccta ttttactttt cctgatagac
     2341 acgtccgctt ctatgaatca gcgcacttat ttgggtacga cgtatctgga cattgctaaa
     2401 ggcgcggttg agatctttat gaagctgcgt gcccgagacc cggctagtag aggcgacagg
     2461 tacatgctag ttacattcga tgatccacca tacggcgtaa aggcgggctg gaaagagaat
     2521 catgccacat tcatgagtga gctgaagaac ctgcaggcat ctggactgac tacactgggt
     2581 cacgctcttc gtgccgcctt cgacctgctc aatctcaaca gacttgtctc tggcatcgac
     2641 aactatgggc agggtcgaaa tccgttcttt ctggagccgt cagtcatcat tacaattaca
     2701 gatggaaaca aactgacaca cagctcggga gtggctgagg agcttcacct gccgctgaat
     2761 tccccattgc ctggtagtga gctaactaag gagccgttcc gatgggacca gagactcttc
     2821 gccctggtcc tgcgtctgcc aggagtggcc gtgcccgaca gtgagcagct gggaagtgtg
     2881 cccactgatg agtctgccat aacccagatg tgtgaggtca ctggaggccg ctcttactgt
     2941 gtaagaaccc aaaggatgct gaaccagtgt cttgagtctc tcgtccaaaa ggtgttgagt
     3001 ggagttgtaa tacattttga gaaaaccggt ccagatccac cagtgattgg tgaagatggt
     3061 cttgtggatc cagctcgtcc tttgacgtcc ttcagtccac agccttggca cagctgccac
     3121 aagctcattt acgtccggcc caaccccaag actggcgtac ctgtgggtca ctggccaatc
     3181 tcagaatcct tctggccaga ccaaaactcc cctactttgc ctcctcgttc tgcccatcct
     3241 gtggtgcggt tttcctgtgt tgactgtgag ccgatggtta tcgacaagct gccttttgac
     3301 aaatatgagc tggagccgtc cccgctcacc cagtacatct tagaaaggaa atctccgcac
     3361 atgtgctggc aggtgtttgt gaactgcagt ggcaaacata gtgatgttgc ccacccgttc
     3421 ggctacctga aggccagcac cacactcacc tgcgtcaacc tcttcgtcat gccttacaac
     3481 tatcctgtcc tgttgccact cctcgatgat ttgtttaaag tgcacaaact aaagccaaac
     3541 ctcaagtggc ggcagtcctt tgaaatgtac ctgaaatcca tgcctccgta ctatctactg
     3601 cctctcaaga aggcgctaag gatgatggga gccccgaatc tcatctctga taacatggat
     3661 tgtggcttga gctacagtgt catttcctac ctgaagaaac tcagccagca ggcaaagatt
     3721 gagtcagacc gtctgattgt gtctgtcggt aagaagcctc ctccagagac gggcatcaag
     3781 gtgaagaacc actcgaatgc cttgtctctg gcacaccgcc gggatttcaa acagctcctg
     3841 cagggcatca cgggtgaggt tcccctccgg ctcatcgaca tgaacttcaa agagtttgct
     3901 ggtttccaga tcgcacttct caacaaggat ttgaagcccc aggcctatag gaatgcttat
     3961 gatattccac gacgaaacct tctagatcaa gtgactcgaa tgcgttccaa ccttttgagg
     4021 acaacgcaaa agctcatcag aggccaggat gatgactctt tgcacagtat tccagtggga
     4081 cagatgggaa attatcagga gtatctgaag atgatgccat cacctctgcg tgagattgat
     4141 ccagaccagc ccaaacgact gcacacattc ggcaacccat tcaaacaaga caagaaggga
     4201 atgatgatcg atgaagccga tgaatttgtg acaggtcctc aaaataagaa gcggggtaac
     4261 acgggggatc taaactcggg tacagctcta aagaggagga ggagtatgtc ccctctgctc
     4321 cgccggcccc aaacccctcc aataataacc aatcatgtat tgggcaaagg gcccacaggg
     4381 actcaaggcc aacaaggaat catcaagccc attccactac acaaaggagc tgaggggaac
     4441 aatgtaggag gcacagaaag caatggcgag cgagtggcag gtactgacgc tggagactgc
     4501 tggcctggag aagtggacgg agaatcagga gaacccgcac ctgtggagga cagagaagac
     4561 gctgcggcac cggacggaga ggaggagata ctagcactgg aaaattgtct tgatgacaga
     4621 tcccctgacc acacgcaaaa ctgtgaggag ctcagtcctc cgggccagga gggggagatg
     4681 gaagtcaatg aaggagacac tccagcacag ggcacgattg tcatgattcc cctggagggg
     4741 agtaacgcag agctgcgcac tcgggttatc aaagaggtcc gcaagcctgg ccgcaattat
     4801 gaggcgatat ttaggctgct ggaggaggtg aaagggcctg tatcagtcca gaggtacttt
     4861 attcatcatg ccatcaaaga ggcagccagg tttaaaaagc gtatgctgat tcagcagctg
     4921 gaaaccgctc tggaggagat tgaagacaga cagatgctac ccgcacaaat caacaatgtc
     4981 cacagcagat agtgcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc
     5041 cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga
     5101 gtttatctga ctaaatctta gtttgtattg tcatgtttta atacaatatg ttatgtttaa
     5161 atatgttttt aataaatttt ataaaataat ttcaactttt attgtaacaa cattgtccat
     5221 ttacacactc ctttcaagcg cgtgggatcg atgctcactc aaaggcggta atacggttat
     5281 ccacagaatc aggggataac gcaggaaaga acatgtgagc catatgggcc catgtgagcc
     5341 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5401 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5461 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5521 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5581 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5641 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5701 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5761 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5821 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5881 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5941 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6001 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6061 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6121 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6181 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6241 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6301 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6361 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6421 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6481 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6541 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6601 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6661 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6721 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6781 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6841 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6901 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6961 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7021 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7081 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7141 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7201 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7261 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7321 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7381 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7441 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7501 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7561 cgttggccga t
//
