LOCUS       sequence-384215-        8439 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses FLAG-tagged Drosophila IntS6 amino acids 101-1284.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8439)
  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 8439)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-IntS6 AA 101-1284
FEATURES             Location/Qualifiers
     source          1..8439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(98..117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      911..1499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1400..1419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1653..1670
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1787..1808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1823..1828,1829..1846,1847..1853)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1861..1877
                     /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     1866..1888
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1885..1901
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1885..1901
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3962..4027
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7620..7661
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7671..7688
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7706..7835
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(7949..7968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8068..8090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8128..8146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8214..8318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8319..8387,8388..8439,1..740)
                     /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"
ORIGIN
        1 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
       61 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      121 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      181 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      241 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      301 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      361 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      421 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      481 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      541 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      601 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      661 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      721 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
      781 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      841 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      901 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      961 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1021 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1081 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1141 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1201 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1261 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1321 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1381 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1441 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1501 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1561 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1621 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1681 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1741 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     1801 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     1861 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttt
     1921 gtcgccggaa cgcagcgaca gagattccaa tgtgtccgta tctttcaggc ttttgccctt
     1981 cagttccaga cgaagcgact ggcgattcgc gtgtggggtc tgcttcaggg tcttgtgaat
     2041 tagggcgcgc agatcgccga tgggcgtggc gccggagggc accttcacct tgccgtacgg
     2101 cttgctgttc ttcgcgttca aaatctccag ctccattttg ctttcggtgc gcttgcaatc
     2161 agtactgtcc aaaatcgaaa atcgccgaac cgtagtgtga ccgtgcgggg ctctgcgaaa
     2221 ataaactttt ttaggtatat ggccacacac ggggaaagca cagtggatta tatgttttaa
     2281 tattataata tgcaggtttt cattacttat ccagatgtaa gcccacttaa agcgatttaa
     2341 caattatttg ccgaaagagt ataaacaaat ttcacttaaa aatggattaa gaaaagctag
     2401 cttgtgtaag attatgcgca gcgttgccag atagctccat ttaaaacact tcaaaaacaa
     2461 taagttttga aaatatatac ataaatagca gtcgttgccg caacgctcaa cacatcacac
     2521 ttttaaaaca ccctttacct acacagaatt actttttaaa tttccagtca agctgcgagt
     2581 ttcaaaatta tagccggtag agaagacagt gctatttcaa aagcaaacta acaagggtct
     2641 taaattccaa aacaccaatc ctaacaagcc ttggactttt gtaagtttag atcaaaggtg
     2701 gcattgcatt caatgtcatg gtaagaagta ggtcgtctag gtagaaatcc tcattcagcc
     2761 ggtcaagtca gtacgagaaa ggtctcaatt tgaaattgtc ttaaaaatat tttattgttt
     2821 tgtactgtgg tgagtttaaa cgaaaaacac aaaaaaaaag tgatacacag aaatcataaa
     2881 aaattttaat acaaggtatt cgtacgtatc aaaaacattt cggcacaatt ttttttctct
     2941 gtactaaagt gttacgaaca ctacggtatt ttttagtgat tttcaacgga caccgaaggt
     3001 atataaacag cgttcgcgaa cggtcgcctt caaaaccaat tgacatttgc agcagcaagt
     3061 acaagtagaa agtaaagcgc aatcagcgaa aaatttatac ttaattgttg gtgattaaag
     3121 tacaattaaa agaacattct cgaaagtcac aagaaacgta agtttttaac tcgctgttac
     3181 caattagtaa taagagcaac aagacgttga gtaatttcaa gaaaaactgc atttcaaggt
     3241 ctttgttcgg ccattttttt ttattcaacg ctctacgtaa ttacaaaata agaaattggc
     3301 agccacgcat cttgttttcc caatgaattg gcatcaaaac gcaaacaaat ctataaataa
     3361 aacttgcgtg ttgattttcg ccaagattta ttggcaaatt gtgaaattcg cagtgacgca
     3421 tttgaaaatt cgagaaatca cgaacgcact cgatcgagca tttgtgtgca tgttattagt
     3481 tagttgttta gttaattgaa gtattttacc aacgaaatcc acttattttt agctgaaata
     3541 gagtaggttg cttaaacaaa gccacgtctg aaaatttctt attgcttgta gttgtgacgt
     3601 caccatatac acacaaaata atgtgtatgc atgcatttca gctgtgtata tatacatgca
     3661 cacactcgca acacgaaaac gatgacgaag caacggaaca aaggtttctc aactaccctt
     3721 tgttccctgt ttcttcgctt tcctttgttc caatattcgt agagggttaa taggggtttc
     3781 tcaacaaagt tggcgtcgat aaataagttt cccattttta ttccccagcc aggaagttag
     3841 tttcaatagt tttgtaattt caacgaaact catttgattt cgtactaatt ttccacatct
     3901 ctattttctg cccgcagaat aatccaaact gcaggtcgac tctagctaga ggaagcttat
     3961 ggactacaaa gaccatgacg gtgattataa agatcatgat atcgattaca aggatgacga
     4021 tgacaaggat ccactagtct taaatctgaa tcgcatgcag tcgggcatcg atacgtacgg
     4081 gcagggcagg tgcccgtttt atctggagcc atcggtcatc attgtgatta cggacggcgg
     4141 tcgctattcg taccggaatg gtgtccatca ggagatcata ctgccgctaa gtaaccaaat
     4201 accgggcaca aagttcacta aggagccatt tcgctgggat cagcgcttgt tttcgcttgt
     4261 cctccgcatg ccgggcaaca agattgacga gcgagtggat ggcaaggtgc cgcatgacga
     4321 ttctcccatc gaacggatgt gcgaggttac cggcggacgt tcatatcgag tgcgaagcca
     4381 ctacgtactc aaccagtgca ttgaaagtct tgtccaaaag gttcagcctg gtgtggtgct
     4441 gcagttcgag cctatgctgc ccaaggaggc tacttccgct accgctggcg aagcggcggg
     4501 tgcgagcacg atatccggca tgggaatacc ctcaacgtcc ggagcgggtc ccgctcccga
     4561 tattgtcttc catccggtaa agaagatgat ctacgtgcag aagcacatca cgcagaaaac
     4621 ctttcccatc ggctattggc cactgccgga gccctattgg ccggactcca aggcgattac
     4681 attgccgcct cgcgacgctc atcccaagct gaaggtcctg acgccggcgg tggacgagcc
     4741 acagctggtg cgcagcttcc ccgtcgacaa gtacgagatt gaaggctgtc cgttaacgct
     4801 gcagattctc aacaagcgcg agatgaacaa gtgctggcag gtgattgtga ccaatggcat
     4861 gcatggattc gagctgccct ttggctacct aaaagcggcg cccaactttt cgcaggtaca
     4921 cctctatgta ctggcctaca attatccagc gttgctgccc attctgcatg acctcattca
     4981 caagtacaat atgagtccgc cgagcgatct gatgtacaaa ttcaacgcct atgtgcgttc
     5041 aatacccccg tattactgcc cgttcctgcg caaggcgctg gtcaacatca atgtgccgta
     5101 tcagctgctg cagtttctgc tgccggagaa tgtggacaac tatctgtcgc caacgatcgc
     5161 caatcagctt aagcacatca agaatacggc caagcaggat caggagaatc tttgcatgaa
     5221 ggtgtacaag cagctgaagc agccgaagcc accatatcgc caagtggaga cgggcaaact
     5281 gttcacaggt gcgccgttac gcagagattt ggtgcggcat ccactgctcc gggatatatt
     5341 ttccaagttg catggggaaa tagatcccgt ggagaattat acgattgtgg tgccgcagcc
     5401 aacgcatcag tcgtcggcca agtcgcatcg caatcccttc gatataccgc ggcgggattt
     5461 ggtggaggaa atagccaaga tgcgtgagac cttactgcga cccgtctcgc tggtggccaa
     5521 ggactcgggc cattgcctgc ccatcgccga gatgggcaac taccaggagt acctcaagaa
     5581 caaggacaat ccgctgcgtg aaatcgagcc gacgaatgtg cggcagcaca tgttcggtaa
     5641 tccgtacaag aagaacaagc acatggtgat ggtggacgag gcggatctga gcgatgtggc
     5701 gccaatgaag tcgccaaatg gcaatggacc gggtggagcg ccacccggct caccttccgg
     5761 atcgggctcg cccacaggac cgggtccctc atctccgggc tcttcgcccg gcggtggttc
     5821 gggtcccggg atgcccggaa tgcccggaat gggcggtgga atgtcaggac tgatgctggg
     5881 cgcaggtggc agtggcggat cctctaaaaa actggacggc acgtctcgtg ggcgaaagag
     5941 gaaagcaggc ccgctgccac gtagctttga attccgtcga tcatctacgg actcgcgatc
     6001 ttcgagctcc ggctccgaat tatccacaac gggtagtgca ccaggatcac cgataccagg
     6061 cgccacgtca tccatatccg gctgcgattc cgccatgggc gctgatggcg gtcccagttc
     6121 cagctgtttc gacgaggaca gcaatagcaa cagcagcttc gtgagctcca catcggaggc
     6181 ctcggccagt gattcaggga tgtcaaactc gcatttggat cccttgcgaa tcagtttcgt
     6241 tttcatcgag gaggagacca gcgatcagga tacgccgctc aatggttatg tgcacaattt
     6301 catcaatggc atcagcgacg atgcagaaac gggcgaaacg gaagcagttc caggcggggc
     6361 atccctgccg ggtgcgtctt cagctaacga accgtcgtca attggagcat caccagcagt
     6421 ttcagccacg ccagcaacat ctgtgatccc cgccagcaat ggcagcgaag tttgctccgc
     6481 cgtcgcggcc ggcagcagca gttccatcag ctccagcgcc agttccccgg gcgttctcta
     6541 cgtgcccctc aacggactga ccacccatgc ggcctacagc agtaatctgg gtggtccgag
     6601 ctcgccgttg gacaacctta gctccctggg tggagcggcg ggtggcggag gatcgggact
     6661 actcggtgtc agcaagccgg gatcgctgcc actggccaat ggttatggac acggtcatag
     6721 ttcagccatt tccccacctt cgccattgtc cttggtgcct ctatcaccat tgggaacaag
     6781 tacgcccgct gccagcagct cgggactatc gtctagttcc gcctacttca gccacaatga
     6841 catcaacgat gcctcggaga tctcgcgcat actgcagacc tgtcacaatg gaaccagcag
     6901 tggctcctcg gtcggagcgg gcgcaagcaa cagcaatttg aatggcaatg gcagcacgga
     6961 gagtggcgga ggagtctcat ccgacgatca tgcctcgctg ggtggcaaca gtttcgatgc
     7021 cctgaaacgg acagcgggta ctgcgggcag cactgcagcg ggaaatcccc actactactg
     7081 ggccagtggt ctcaatcatc acaacaataa caacagcagc gccgctggag cagctagcgg
     7141 ttcgacgttg agcaacaatc acaaccaccg gggtcacaac aacagcagcc ccaacaaact
     7201 ggaggtgaag atcaactcca gctgcggctc ctcaccgacg cacaacaacg ggggcatggg
     7261 ctcaccggga cagagcctgc cgctcatttt caccgaggag cagcgcgaag cggcccggtt
     7321 gcacaacgtc gaactacgat tgcagatctt ccgtgatata cggcgacccg gccgagacta
     7381 cagccagctg ctggagcacc tgaatctggt caagggcgac caggatatgc agtcggattt
     7441 cgtggacatg tgtatcgtgg aatcgctgcg cttccgccgc catcgcatgg cgtccagcat
     7501 acaggagtgg tgggatcgca agcagcagtt aactgccgtg ggaaccaatg aagcgacccc
     7561 cagcgcggag caggccgtcg ccaagagtta actcgagtct agagggcccg cggttcgaag
     7621 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc
     7681 atcaccattg agtttatctg actaaatctt agtttgtatt gtcatgtttt aatacaatat
     7741 gttatgttta aatatgtttt taataaattt tataaaataa tttcaacttt tattgtaaca
     7801 acattgtcca tttacacact cctttcaagc gcgtgggatc gatgctcact caaaggcggt
     7861 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag ccatatgggc
     7921 ccatgtgagc catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7981 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     8041 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     8101 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     8161 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     8221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     8281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     8341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     8401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcac
//
