LOCUS       sequence-384311-        6648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila Pp2A-29B protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6648)
  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 6648)
  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-Pp2A-29B
FEATURES             Location/Qualifiers
     source          1..6648
                     /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             6150..6191
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6201..6218
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6236..6365
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(6479..6498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6598..6620
                     /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 tccagcagca agcgacaaat cggtcgacga
     4381 ttcactatat cccattgcgg ttctaatcga tgaactgaaa aacgaggacg ttcagcttcg
     4441 gttgaactcc atcaagaaac tgtccaccat tgcactcgct ttgggcgagg agcgcacacg
     4501 gtccgagttg attcccttcc tcaccgagac catatacgat gaggacgagg tactgctggc
     4561 cctggccgac caactgggca actttactag tctcgttggt gggccagagt ttgccatgta
     4621 cttgattccg cccctcgaga gtttggccac cgtagaggaa accgtggtgc gagacaaggc
     4681 tgtggaatct ctacgcaccg tggccgctga gcacagcgcc caggatttgg agatccatgt
     4741 ggtgccgaca ctgcagcgat tggtttccgg tgactggttc acctcacgca cctctgcctg
     4801 cggcctcttc tcggtctgct atccacgcgt cacacagcca gtgaaggccg agctgcgcgc
     4861 caacttccga aagctctgcc aggatgagac acccatggtg cgccgtgcag cggccaacaa
     4921 gctgggcgag tttgccaagg tcgttgagac ggagtatctg aagtccgatt tgattcccaa
     4981 ctttgtccag ctggcacagg atgatcagga ctctgtccgt ctgctggctg tagaggcatg
     5041 cgtaagcatt gcccagctgc tgccacagga tgatgtagag cacctggttc tgcccacgct
     5101 gcgccagtgc gccagcgact cttcctggag ggtgcgttac atggtggccg agaagtttgt
     5161 tgatctgcaa aaggctgtgg gcccagagat tactagggtg gacttggtgc ctgccttcca
     5221 gtacttgctc aaggatgccg aggccgaggt tcgcgctgca gtggccacca aggtgaagga
     5281 cttctgcgcc aatctggaca aggtcaacca ggtgcaaatc atccttagtt ccattttgcc
     5341 ctatgtccgc gatcttgtct cggaccccaa tcctcatgtg aagtcagctc tggcctcagt
     5401 gatcatgggc ttgagtccca tgctgggcgc ctatcagact gtggagcaat tgctccccct
     5461 gttccttatt caactcaagg atgagtgccc agaagtgcgc ctaaacatca tctcaaacct
     5521 ggattgcgtt aacgacgtca tcggtatcca gcaactgtca cagtcgcttc tgcccgccat
     5581 cgtcgagctg gccgaggact ccaagtggcg tgtgcgtcta gccatcatcg agtacatgcc
     5641 tgctctggcc ggtcagttgg gtcaggaatt ctttgaccaa aaactgcgcg gtctctgcat
     5701 gggatggctc aacgatcacg tgtacgccat tcgtgaggca gccaccctca acatgaagaa
     5761 gctcgtcgag cagttcggag ctccctgggc cgaacaggcc ataattccaa tgattctggt
     5821 tatgtcgcgc aacaagaact atttgcacag aatgacttgc ttgttctgcc tgaatgtttt
     5881 ggcagaggtc tgcggcacag atatcaccac caagttgctg ctgcccacag ttctcctgct
     5941 tgccgctgat cccgttgcca atgttcgttt caacgtggca aagaccctgc agaagatctc
     6001 gcccttcctg gaggccagcg tcattgatgc ccaagtaaag cccacactcg acaaactgaa
     6061 cacagacaca gatgtcgatg tcaagcattt tgctgcacag gccattgccg gcatagctgc
     6121 agcgtaatct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     6181 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttatctg actaaatctt
     6241 agtttgtatt gtcatgtttt aatacaatat gttatgttta aatatgtttt taataaattt
     6301 tataaaataa tttcaacttt tattgtaaca acattgtcca tttacacact cctttcaagc
     6361 gcgtgggatc gatgctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6421 cgcaggaaag aacatgtgag ccatatgggc ccatgtgagc catatggtgc actctcagta
     6481 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     6541 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6601 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
