LOCUS       sequence-384213-        6223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila tws protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  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 6223)
  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-tws
FEATURES             Location/Qualifiers
     source          1..6223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4283..4348
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5725..5766
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5776..5793
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5811..5940
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(6054..6073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6173..6195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt tgtcgccgga acgcagcgac agagattcca atgtgtccgt
     2281 atctttcagg cttttgccct tcagttccag acgaagcgac tggcgattcg cgtgtggggt
     2341 ctgcttcagg gtcttgtgaa ttagggcgcg cagatcgccg atgggcgtgg cgccggaggg
     2401 caccttcacc ttgccgtacg gcttgctgtt cttcgcgttc aaaatctcca gctccatttt
     2461 gctttcggtg cgcttgcaat cagtactgtc caaaatcgaa aatcgccgaa ccgtagtgtg
     2521 accgtgcggg gctctgcgaa aataaacttt tttaggtata tggccacaca cggggaaagc
     2581 acagtggatt atatgtttta atattataat atgcaggttt tcattactta tccagatgta
     2641 agcccactta aagcgattta acaattattt gccgaaagag tataaacaaa tttcacttaa
     2701 aaatggatta agaaaagcta gcttgtgtaa gattatgcgc agcgttgcca gatagctcca
     2761 tttaaaacac ttcaaaaaca ataagttttg aaaatatata cataaatagc agtcgttgcc
     2821 gcaacgctca acacatcaca cttttaaaac accctttacc tacacagaat tactttttaa
     2881 atttccagtc aagctgcgag tttcaaaatt atagccggta gagaagacag tgctatttca
     2941 aaagcaaact aacaagggtc ttaaattcca aaacaccaat cctaacaagc cttggacttt
     3001 tgtaagttta gatcaaaggt ggcattgcat tcaatgtcat ggtaagaagt aggtcgtcta
     3061 ggtagaaatc ctcattcagc cggtcaagtc agtacgagaa aggtctcaat ttgaaattgt
     3121 cttaaaaata ttttattgtt ttgtactgtg gtgagtttaa acgaaaaaca caaaaaaaaa
     3181 gtgatacaca gaaatcataa aaaattttaa tacaaggtat tcgtacgtat caaaaacatt
     3241 tcggcacaat tttttttctc tgtactaaag tgttacgaac actacggtat tttttagtga
     3301 ttttcaacgg acaccgaagg tatataaaca gcgttcgcga acggtcgcct tcaaaaccaa
     3361 ttgacatttg cagcagcaag tacaagtaga aagtaaagcg caatcagcga aaaatttata
     3421 cttaattgtt ggtgattaaa gtacaattaa aagaacattc tcgaaagtca caagaaacgt
     3481 aagtttttaa ctcgctgtta ccaattagta ataagagcaa caagacgttg agtaatttca
     3541 agaaaaactg catttcaagg tctttgttcg gccatttttt tttattcaac gctctacgta
     3601 attacaaaat aagaaattgg cagccacgca tcttgttttc ccaatgaatt ggcatcaaaa
     3661 cgcaaacaaa tctataaata aaacttgcgt gttgattttc gccaagattt attggcaaat
     3721 tgtgaaattc gcagtgacgc atttgaaaat tcgagaaatc acgaacgcac tcgatcgagc
     3781 atttgtgtgc atgttattag ttagttgttt agttaattga agtattttac caacgaaatc
     3841 cacttatttt tagctgaaat agagtaggtt gcttaaacaa agccacgtct gaaaatttct
     3901 tattgcttgt agttgtgacg tcaccatata cacacaaaat aatgtgtatg catgcatttc
     3961 agctgtgtat atatacatgc acacactcgc aacacgaaaa cgatgacgaa gcaacggaac
     4021 aaaggtttct caactaccct ttgttccctg tttcttcgct ttcctttgtt ccaatattcg
     4081 tagagggtta ataggggttt ctcaacaaag ttggcgtcga taaataagtt tcccattttt
     4141 attccccagc caggaagtta gtttcaatag ttttgtaatt tcaacgaaac tcatttgatt
     4201 tcgtactaat tttccacatc tctattttct gcccgcagaa taatccaaac tgcaggtcga
     4261 ctctagctag aggaagctta tggactacaa agaccatgac ggtgattata aagatcatga
     4321 tatcgattac aaggatgacg atgacaagga tccactagtc gccggtaatg gagaggcgtc
     4381 ttggtgcttc tcacagatta aaggcgccct agacgatgat gtcacggatg cagacatcat
     4441 atcctgcgtg gaattcaatc acgatggcga gctgctggcc actggtgaca agggcggtcg
     4501 cgtcgtcatc tttcagcgtg atcctgcctc aaaagccgcc aatccgagac gcggcgaata
     4561 caatgtctat tcgacattcc aatcgcacga gcccgaattc gactacctca agtccctgga
     4621 gattgaggag aagataaaca agatccggtg gctgcaacaa aagaatcccg tgcactttct
     4681 gctctcgacc aacgacaaga cagtcaaatt gtggaaggtc agtgagcgtg acaagtcgtt
     4741 cggcggctac aacacgaagg aggagaacgg actgatccgg gatccacaga atgtaacggc
     4801 cctccgagtg ccatccgtga agcagatacc actgctggtg gaggcctccc cacggcgcac
     4861 cttcgccaat gcacacacct atcacatcaa ttcaataagc gttaattcgg atcaggagac
     4921 cttcttgtcg gccgacgacc tgcggatcaa tctgtggcac ttggaggtgg tcaaccagag
     4981 ctacaacatt gtcgacatca agccaaccaa catggaggag ctaacagagg tgatcaccgc
     5041 ggccgaattc catccgaccg agtgcaatgt gtttgtctac tcgagctcta agggcacaat
     5101 acgattatgt gatatgcgtt cggcggcgct atgcgaccgg cacagcaaac agttcgagga
     5161 gcccgagaat ccaacgaatc gcagcttttt cagcgaaata atcagctcca tcagcgatgt
     5221 gaagctaagc aactcgggtc gctacatgat ctccagggat tacttgagca tcaaagtgtg
     5281 ggatctgcat atggaaacaa agcccattga gacctatccg gttcatgaat acctgcgcgc
     5341 caagctgtgc tcgctgtacg agaatgactg catcttcgac aagttcgagt gctgttggaa
     5401 cggcaaggac agctcaataa tgaccggaag ctacaacaac ttcttccgcg tcttcgatcg
     5461 caactcgaaa aaggatgtga cgctagaggc gtccagggac atcatcaaac cgaaaacggt
     5521 gcttaagcca cggaaagttt gcactggcgg caaacgaaag aaggatgaga tcagcgtgga
     5581 ctgtctagat ttcaacaaga agatcctgca caccgcctgg caccccgaag aaaatatcat
     5641 cgccgtggct gcgaccaata acctcttcat atttcaggat aaattttagg cggccgctcg
     5701 agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat
     5761 tctacgcgta ccggtcatca tcaccatcac cattgagttt atctgactaa atcttagttt
     5821 gtattgtcat gttttaatac aatatgttat gtttaaatat gtttttaata aattttataa
     5881 aataatttca acttttattg taacaacatt gtccatttac acactccttt caagcgcgtg
     5941 ggatcgatgc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6001 gaaagaacat gtgagccata tgggcccatg tgagccatat ggtgcactct cagtacaatc
     6061 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     6121 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     6181 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
