LOCUS       sequence-389999-        7542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged human IntS6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7542)
  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 7542)
  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-Human IntS6
FEATURES             Location/Qualifiers
     source          1..7542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    112..133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(148..153,154..171,172..178)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    186..202
                     /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     191..213
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     210..226
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     210..226
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2287..2352
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5048..5089
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5099..5116
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5134..5263
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(5377..5396)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5496..5518
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5556..5574)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5642..5746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5747..5815,5816..6607)
                     /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(5965..5984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6778..7366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7267..7286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7520..7537
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
       61 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      121 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      181 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      241 gctttgtcgc cggaacgcag cgacagagat tccaatgtgt ccgtatcttt caggcttttg
      301 cccttcagtt ccagacgaag cgactggcga ttcgcgtgtg gggtctgctt cagggtcttg
      361 tgaattaggg cgcgcagatc gccgatgggc gtggcgccgg agggcacctt caccttgccg
      421 tacggcttgc tgttcttcgc gttcaaaatc tccagctcca ttttgctttc ggtgcgcttg
      481 caatcagtac tgtccaaaat cgaaaatcgc cgaaccgtag tgtgaccgtg cggggctctg
      541 cgaaaataaa cttttttagg tatatggcca cacacgggga aagcacagtg gattatatgt
      601 tttaatatta taatatgcag gttttcatta cttatccaga tgtaagccca cttaaagcga
      661 tttaacaatt atttgccgaa agagtataaa caaatttcac ttaaaaatgg attaagaaaa
      721 gctagcttgt gtaagattat gcgcagcgtt gccagatagc tccatttaaa acacttcaaa
      781 aacaataagt tttgaaaata tatacataaa tagcagtcgt tgccgcaacg ctcaacacat
      841 cacactttta aaacaccctt tacctacaca gaattacttt ttaaatttcc agtcaagctg
      901 cgagtttcaa aattatagcc ggtagagaag acagtgctat ttcaaaagca aactaacaag
      961 ggtcttaaat tccaaaacac caatcctaac aagccttgga cttttgtaag tttagatcaa
     1021 aggtggcatt gcattcaatg tcatggtaag aagtaggtcg tctaggtaga aatcctcatt
     1081 cagccggtca agtcagtacg agaaaggtct caatttgaaa ttgtcttaaa aatattttat
     1141 tgttttgtac tgtggtgagt ttaaacgaaa aacacaaaaa aaaagtgata cacagaaatc
     1201 ataaaaaatt ttaatacaag gtattcgtac gtatcaaaaa catttcggca caattttttt
     1261 tctctgtact aaagtgttac gaacactacg gtatttttta gtgattttca acggacaccg
     1321 aaggtatata aacagcgttc gcgaacggtc gccttcaaaa ccaattgaca tttgcagcag
     1381 caagtacaag tagaaagtaa agcgcaatca gcgaaaaatt tatacttaat tgttggtgat
     1441 taaagtacaa ttaaaagaac attctcgaaa gtcacaagaa acgtaagttt ttaactcgct
     1501 gttaccaatt agtaataaga gcaacaagac gttgagtaat ttcaagaaaa actgcatttc
     1561 aaggtctttg ttcggccatt tttttttatt caacgctcta cgtaattaca aaataagaaa
     1621 ttggcagcca cgcatcttgt tttcccaatg aattggcatc aaaacgcaaa caaatctata
     1681 aataaaactt gcgtgttgat tttcgccaag atttattggc aaattgtgaa attcgcagtg
     1741 acgcatttga aaattcgaga aatcacgaac gcactcgatc gagcatttgt gtgcatgtta
     1801 ttagttagtt gtttagttaa ttgaagtatt ttaccaacga aatccactta tttttagctg
     1861 aaatagagta ggttgcttaa acaaagccac gtctgaaaat ttcttattgc ttgtagttgt
     1921 gacgtcacca tatacacaca aaataatgtg tatgcatgca tttcagctgt gtatatatac
     1981 atgcacacac tcgcaacacg aaaacgatga cgaagcaacg gaacaaaggt ttctcaacta
     2041 ccctttgttc cctgtttctt cgctttcctt tgttccaata ttcgtagagg gttaataggg
     2101 gtttctcaac aaagttggcg tcgataaata agtttcccat ttttattccc cagccaggaa
     2161 gttagtttca atagttttgt aatttcaacg aaactcattt gatttcgtac taattttcca
     2221 catctctatt ttctgcccgc agaataatcc aaactgcagg tcgactctag ctagaggaag
     2281 cttatggact acaaagacca tgacggtgat tataaagatc atgatatcga ttacaaggat
     2341 gacgatgaca aggatccact agtccccatc ttactgttcc tgatagacac gtctgcctct
     2401 atgaaccagc gcagccatct gggcaccacc tacctggaca cggccaaagg cgcggtagag
     2461 accttcatga agctccgtgc ccgggaccct gccagcagag gagacaggta tatgctggtc
     2521 actttcgaag agccgcccta tgctatcaag gctggatgga aagaaaacca tgcaacgttt
     2581 atgaatgaat tgaaaaacct tcaggctgaa ggacttacga ctcttggcca atccctaagg
     2641 acagcttttg atttattaaa tttaaataga ttagtaactg gcatagacaa ctatgggcag
     2701 ggaagaaacc cttttttctt ggagccagca ataattatca caattactga tgggagcaag
     2761 ttgactacca ccagtggagt ccaggatgag cttcatttac ctcttaattc tcctttgcct
     2821 ggaagtgaat tgaccaagga accttttcgt tgggatcaga gactctttgc attagtgttg
     2881 cggttgcctg gcaccatgtc agtagaatca gaacagttga caggtgtgcc tttagatgac
     2941 tctgcaatca caccaatgtg tgaagtgaca ggcggccgtt catattctgt gtgttctcca
     3001 agaatgctta atcagtgtct ggagtccttg gtgcagaaag tacaaagtgg ggtggtaata
     3061 aactttgaaa aagcaggacc agatccttcc cctgtagaag atgggcagcc agatatatca
     3121 aggccttttg gatctcagcc ttggcatagc tgtcacaaac tcatatatgt cagaccaaat
     3181 cctaaaactg gggttcctat aggtcattgg cctgttccag agtctttttg gccagatcaa
     3241 aattcgccaa cactaccacc tcgtacatct catcctgtag tgaagttttc ctgtacagac
     3301 tgtgaaccaa tggttattga taaacttcct tttgacaaat atgagttgga accttcacca
     3361 ctgactcaat ttatcctgga aaggaaatct cctcaaacat gttggcaggt gtacgtgagc
     3421 aatagtgcaa aatacagtga acttggtcat ccttttggtt acttgaaagc cagtacagca
     3481 ctgaactgtg tcaacttatt tgtgatgcct tacaattatc cagtccttct tcccctctta
     3541 gatgacttgt ttaaagtgca taaagcaaaa ccaacattga agtggagaca gtcatttgaa
     3601 agttatttga agacaatgcc tccctactat cttgggccct tgaagaaagc tgttaggatg
     3661 atgggagcac ctaacctaat agcagacagt atggaatatg gacttagtta cagtgtcatt
     3721 tcatacctca aaaaactgag tcaacaggcc aaaatagaat ctgatcgagt cattggatct
     3781 gtaggcaaaa aagtagtaca ggagactgga ataaaagtcc ggagccgatc acatggttta
     3841 tcaatggcat ataggaaaga ttttcaacaa ctcctccagg gaatttcaga ggatgtccct
     3901 cacagactgc tagaccttaa tatgaaggaa tacactgggt tccaagttgc tttgctgaat
     3961 aaggatttga agccacagac atttagaaat gcttatgaca taccaagacg aaatcttttg
     4021 gatcacttaa caagaatgag atctaatctt ttgaagagca ctcgcagatt tctgaaagga
     4081 caggacgaag atcaagtgca cagtgttcct atagcacaaa tggggaacta ccaggaatac
     4141 ctcaagcaag taccttctcc actaagagaa cttgatcctg atcagccacg aaggttgcat
     4201 acatttggca acccctttaa gctggataag aagggtatga tgatagatga agcagatgaa
     4261 tttgtggctg gacctcaaaa taaacataaa cgacccggag aaccaaatat gcaagggatc
     4321 cctaaaagac gtcggtgtat gtctccacta ctaagaggca gacagcagaa tcctgttgta
     4381 aacaatcata ttgggggaaa aggaccacct gcacctacaa ctcaagcaca gccagatctt
     4441 attaaacctc ttcctcttca taaaatttca gaaaccacta atgattcgat aatacatgat
     4501 gtggttgaaa atcatgttgc agaccaactt tcatcagaca ttacaccaaa tgctatggat
     4561 acggaatttt cagcatcttc tccagccagt ttactggaac ggccaaccaa tcatatggag
     4621 gctcttggtc atgaccattt aggaaccaat gacctcactg ttggtggatt tttagaaaat
     4681 catgaggagc caagagataa agaacaatgt gctgaagaga acataccagc atcttcactc
     4741 aacaaaggaa agaaattgat gcattgcaga agccatgaag aggtcaatac tgaactaaaa
     4801 gcacaaataa tgaaagagat ccgaaagcca ggaagaaaat atgaaagaat cttcacttta
     4861 ctgaagcatg tgcaaggcag tttacaaaca agactaatat ttttacaaaa tgtcattaaa
     4921 gaagcatcaa ggtttaaaaa acgaatgcta atagaacaac tggagaactt cttggatgaa
     4981 attcatcgaa gagccaatca gatcaaccat attaatagca attaatctag agggcccgcg
     5041 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     5101 tcatcaccat caccattgag tttatctgac taaatcttag tttgtattgt catgttttaa
     5161 tacaatatgt tatgtttaaa tatgttttta ataaatttta taaaataatt tcaactttta
     5221 ttgtaacaac attgtccatt tacacactcc tttcaagcgc gtgggatcga tgctcactca
     5281 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagcc
     5341 atatgggccc atgtgagcca tatggtgcac tctcagtaca atctgctctg atgccgcata
     5401 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5461 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5521 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5581 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5641 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5701 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5761 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5821 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5881 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5941 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6001 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6061 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6121 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6181 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6241 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6301 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6361 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6421 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6481 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6541 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6601 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6661 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6721 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6781 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6841 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6901 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6961 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7021 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7081 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7141 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7201 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7261 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7321 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7381 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7441 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7501 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gc
//
