LOCUS       sequence-390000-        7464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged human IntS6L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7464)
  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 7464)
  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 IntS6L
FEATURES             Location/Qualifiers
     source          1..7464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    32..53
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(68..73,74..91,92..98)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    106..122
                     /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     111..133
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     130..146
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     130..146
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2207..2272
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4890..4931
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4941..4958
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4976..5105
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(5219..5238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5338..5360
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5398..5416)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5484..5588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5589..5657,5658..6449)
                     /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(5807..5826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6620..7208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7109..7128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7362..7379
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
       61 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      121 aatttcacac aggaaacagc tatgaccatg attacgccaa gctttgtcgc cggaacgcag
      181 cgacagagat tccaatgtgt ccgtatcttt caggcttttg cccttcagtt ccagacgaag
      241 cgactggcga ttcgcgtgtg gggtctgctt cagggtcttg tgaattaggg cgcgcagatc
      301 gccgatgggc gtggcgccgg agggcacctt caccttgccg tacggcttgc tgttcttcgc
      361 gttcaaaatc tccagctcca ttttgctttc ggtgcgcttg caatcagtac tgtccaaaat
      421 cgaaaatcgc cgaaccgtag tgtgaccgtg cggggctctg cgaaaataaa cttttttagg
      481 tatatggcca cacacgggga aagcacagtg gattatatgt tttaatatta taatatgcag
      541 gttttcatta cttatccaga tgtaagccca cttaaagcga tttaacaatt atttgccgaa
      601 agagtataaa caaatttcac ttaaaaatgg attaagaaaa gctagcttgt gtaagattat
      661 gcgcagcgtt gccagatagc tccatttaaa acacttcaaa aacaataagt tttgaaaata
      721 tatacataaa tagcagtcgt tgccgcaacg ctcaacacat cacactttta aaacaccctt
      781 tacctacaca gaattacttt ttaaatttcc agtcaagctg cgagtttcaa aattatagcc
      841 ggtagagaag acagtgctat ttcaaaagca aactaacaag ggtcttaaat tccaaaacac
      901 caatcctaac aagccttgga cttttgtaag tttagatcaa aggtggcatt gcattcaatg
      961 tcatggtaag aagtaggtcg tctaggtaga aatcctcatt cagccggtca agtcagtacg
     1021 agaaaggtct caatttgaaa ttgtcttaaa aatattttat tgttttgtac tgtggtgagt
     1081 ttaaacgaaa aacacaaaaa aaaagtgata cacagaaatc ataaaaaatt ttaatacaag
     1141 gtattcgtac gtatcaaaaa catttcggca caattttttt tctctgtact aaagtgttac
     1201 gaacactacg gtatttttta gtgattttca acggacaccg aaggtatata aacagcgttc
     1261 gcgaacggtc gccttcaaaa ccaattgaca tttgcagcag caagtacaag tagaaagtaa
     1321 agcgcaatca gcgaaaaatt tatacttaat tgttggtgat taaagtacaa ttaaaagaac
     1381 attctcgaaa gtcacaagaa acgtaagttt ttaactcgct gttaccaatt agtaataaga
     1441 gcaacaagac gttgagtaat ttcaagaaaa actgcatttc aaggtctttg ttcggccatt
     1501 tttttttatt caacgctcta cgtaattaca aaataagaaa ttggcagcca cgcatcttgt
     1561 tttcccaatg aattggcatc aaaacgcaaa caaatctata aataaaactt gcgtgttgat
     1621 tttcgccaag atttattggc aaattgtgaa attcgcagtg acgcatttga aaattcgaga
     1681 aatcacgaac gcactcgatc gagcatttgt gtgcatgtta ttagttagtt gtttagttaa
     1741 ttgaagtatt ttaccaacga aatccactta tttttagctg aaatagagta ggttgcttaa
     1801 acaaagccac gtctgaaaat ttcttattgc ttgtagttgt gacgtcacca tatacacaca
     1861 aaataatgtg tatgcatgca tttcagctgt gtatatatac atgcacacac tcgcaacacg
     1921 aaaacgatga cgaagcaacg gaacaaaggt ttctcaacta ccctttgttc cctgtttctt
     1981 cgctttcctt tgttccaata ttcgtagagg gttaataggg gtttctcaac aaagttggcg
     2041 tcgataaata agtttcccat ttttattccc cagccaggaa gttagtttca atagttttgt
     2101 aatttcaacg aaactcattt gatttcgtac taattttcca catctctatt ttctgcccgc
     2161 agaataatcc aaactgcagg tcgactctag ctagaggaag cttatggact acaaagacca
     2221 tgacggtgat tataaagatc atgatatcga ttacaaggat gacgatgaca aggatccact
     2281 agtccccatc ctgctgttcc tcatagacac gtccgcctct atgaaccagc gcactgacct
     2341 gggcacctct tatttggaca ttgccaaagg cgctgtggag ttattcttga agctgcgcgc
     2401 ccgggacccg gccagccgtg gagacaggta catgctggtc acctacgacg aacccccgta
     2461 ctgcatcaag gctggttgga aggaaaatca tgcaacattc atgagcgaac taaaaaatct
     2521 tcaggcttct ggactgacta ctctcggtca ggctctaaga tcctcatttg atttgttaaa
     2581 tctcaataga ttaatatctg gaatagacaa ttatggacag gggagaaatc catttttttt
     2641 agaaccatct attttaatta ccatcacaga tggaaacaag ttaacaagta ctgctggtgt
     2701 tcaagaagag ctccatcttc ctttgaattc ccctctgcct ggaagtgaac taaccaaaga
     2761 accttttcgt tgggatcaaa ggttatttgc cctggtgttg cgtttgcctg gagtggcttc
     2821 taccgaacca gagcaactag ggagcgtacc aactgatgaa tctgccatca cacagatgtg
     2881 tgaagtcaca ggaggtcgct cctactgtgt gagaacacaa agaatgttga atcaatgttt
     2941 agaatctcta gttcaaaaag ttcagagtgg tgtagttatt aattttgaaa aaacaggacc
     3001 agatccactt cctattggag aagatggact tatggattca tccaggccaa gcaattcatt
     3061 tgctgctcag ccatggcata gttgtcataa actcatttat gtacgaccta actctaaaac
     3121 tggtgttcct gttggacatt ggccaattcc agaatctttt tggccagatc agaatttacc
     3181 ttcactacct ccacgaacat ctcatcctgt tgtgaggttc tcctgtgtag attgtgagcc
     3241 aatggtaata gacaaacttc cttttgacaa atatgaactt gaaccttcgc ccttaactca
     3301 gtatatcttg gaacgaaagt ctccccatac ctgctggcag gtatttgtta ctagcagtgg
     3361 aaagtacaat gaacttggat atccatttgg ttatttaaaa gccagtacaa ctttaacttg
     3421 tgtaaacctc tttgtgatgc cttacaacta cccagtttta cttcctcttt tagatgactt
     3481 gtttaaagtt cacaagctta agccaaatct gaagtggcga caggcttttg acagctactt
     3541 aaaaactctg cctccatact acctattaac caaactagag tcagaacgaa tactagcatc
     3601 agtggggaag aaacctcccc aggaaattgg aattaaagtg aaaaatcatt ctggaggtgg
     3661 catgtccttg actcacaata aaaattttag aaaactattg aaagaaatca caggggaaac
     3721 tgcacttaga ctgacagaat tgaacaccaa agaatttgct ggcttccaaa ttgggctctt
     3781 aaacaaggat ttgaaacctc agacatacag aaatgcttat gatattcccc gtagaggtct
     3841 tttagaccag ctgaccagaa tgagatccaa tctgctgaaa acgcacaagt ttattgttgg
     3901 acaagatgaa gattcccttc atagtgttcc agttgcacaa atgggtaact atcaggaata
     3961 tctgaagaca ttggcttctc cactgcgaga gattgatcca gaccaaccca aaagactgca
     4021 tacttttggc aatccgttta aacaagataa gaagggaatg atgattgatg aagcagatga
     4081 gtttgtagca gggccacaaa acaaagtgaa acgtccaggg gaacccaaca gtcctatgtc
     4141 atctaagaga aggcggagta tgtccctgct gttgaggaaa ccacaaacac cacctactgt
     4201 aactaaccat gtgggcggaa agggaccacc ctcagcctcg tggttcccat cttatccaaa
     4261 cctcataaaa cccacccttg tacatacaga tgctactatc attcacgatg gccatgagga
     4321 gaagatggaa aatggtcaga tcacacctga tggcttcctg tcaaaatctg ctccatcaga
     4381 gcttataaat atgacaggag atcttatgcc acccaaccaa gtggattctc tgtctgacga
     4441 cttcacaagt ctcagcaaag atgggctgat tcaaaaacct ggtagtaacg catttgtagg
     4501 aggagccaaa aactgcagtc tctccgtaga tgaccaaaaa gacccagtag catctacttt
     4561 gggagctatg ccaaatacat tacaaatcac tcctgctatg gcacaaggaa tcaatgctga
     4621 tataaaacat caattaatga aggaagttcg aaagtttggt cgaaaatatg aaagaatttt
     4681 cattttgctt gaagaagtgc aaggacctct ggagatgaag aaacagtttg ttgaatttac
     4741 catcaaggaa gccgcaaggt ttaaaagacg agtcctaatt cagtaccttg agaaggtact
     4801 agaaaaaata aattcccacc accttcacaa caacattagt cacatcaaca gcagatcatc
     4861 atgttaatct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     4921 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttatctg actaaatctt
     4981 agtttgtatt gtcatgtttt aatacaatat gttatgttta aatatgtttt taataaattt
     5041 tataaaataa tttcaacttt tattgtaaca acattgtcca tttacacact cctttcaagc
     5101 gcgtgggatc gatgctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5161 cgcaggaaag aacatgtgag ccatatgggc ccatgtgagc catatggtgc actctcagta
     5221 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5281 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5341 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5401 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5461 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5521 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5581 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5641 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5701 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5761 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5821 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5881 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5941 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6001 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6061 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6121 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6181 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6241 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6301 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6361 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6421 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6481 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6541 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6601 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6661 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6721 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6781 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6841 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6901 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6961 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7021 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7081 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7141 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7201 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7261 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     7321 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     7381 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     7441 gcagctggca cgacaggttt cccg
//
