LOCUS       sequence-383958-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila cka protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  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 7084)
  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-cka
FEATURES             Location/Qualifiers
     source          1..7084
                     /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"
     regulatory      6309..6318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6586..6627
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6637..6654
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6672..6801
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     complement(6915..6934)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7034..7056
                     /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 ggcaccaatt cgggagccac
     4381 cgctggcata aacaacaagc cggttggcgg tgcaacagga gccggcgtcc ttgtaggcgg
     4441 cggtgtgggc ggtgccaatt cctcgatcgg cggtgtcctg tcgaacagcc tgggcggtgg
     4501 cggcagcggc ggtctgagca tcagcggcct caacgctggt ggacagaacg ccaatgtggg
     4561 cggaatgggc aacgttggcg gcgacgacgg cggaaacggg atggtgggcg gcggtgtaaa
     4621 taaccagcag gccacaacgc cccaatacac aataccgggc atcttgcact tcatccagca
     4681 cgagtggtcg cgcttcgagc tggagcgatc acagtgggac gtggacaggg ccgaattgca
     4741 ggctcgcatt gctatgctgc ttggagagcg caagtgcttg gaaagcctca aatcggattt
     4801 gacgcgccgc atcaagatgc tggagtacgc actgcgccag gagcgtgcga aattctatcg
     4861 tttaaaatac ggcaccgatc cgccgcagct caatgagttc aagccatcga acgaggatgc
     4921 cggactggcc ggtgaggtgg ccactgactc ggaggtgccc tacagcagtg tctcgaacac
     4981 tacgtggcgg caggggcgtc agatgctgcg acagtatctg gccgagattg gctacacaga
     5041 caacatcatc gatgtgcgtt caaatcgggt gcgttcgatc cttggcctaa acaataatgc
     5101 cgagcacgac ggcagcggcg gaggactggg tggtggactt ggcggcggca ccggcggcga
     5161 gaaccttagc ccaaacatca atggaaacga gagcaacaaa cgcgcatcgg agacggaagg
     5221 ccgtcatact cccgctaaaa aagtgcaaca atcgatcgat gaaataatcg tggacaccga
     5281 ggcagctgtg atggcgaact ttgagttcct gggcgccacc gagatgtccg acgacgacga
     5341 gatatctgac gatctggaaa tggtggccac cgacaatgac gacacagacg taaagctggc
     5401 caagcgcgcc aagtctggca aggatatgct caccgaggag gtggacggct cgttgggact
     5461 gggtgagctt gcccagctga cggtgaataa cgaatctgat ggcgcctacg atgcgaactc
     5521 caaggatgga actggcggca gtgccggcgg tgctggttac cgcaaaacct ggaacgccaa
     5581 gtacacgctt cgctcgcact ttgacggagt acgatcactt atctttcacc cagaggagcc
     5641 ggtgctgatc acggcttccg aggatcatac gcttaagctg tggaaccttc agaagacggt
     5701 gcaggccaaa aagtcggcca gcttggatgt ggaaccgctg tatacgttca gggcccatac
     5761 gggtccagtt ctgtgcctgg gcatgtcgtc cagcggcgag acatgctatt cgggcgggct
     5821 ggacggtaac attgaatgct ggcaattgcc gtcgccgaat atcgatccgt acgattgcta
     5881 tgatccgaac gtgcattccg gcacgctcga ggggcatacg gatgccgtgt ggggccttac
     5941 caccatgcaa agcaacattg tgtcatgctc agcggatgga acggttaagc tgtggtcgcc
     6001 atacaataag gaaccgctgc tgcgcacata cacagcctcc gaggcagagg gtgtaccctc
     6061 atccgttgac tttgttcgca atgaggttga ccatatcgtg gtggcctaca acagtgccca
     6121 ctgcatcgtg tacgatacgg agacgggcaa gcaggtggtg cgtctggagg cggcgcagga
     6181 gatgtctggc aacacgggga agttcattaa caaggtggtg tcacacccaa cgctacccat
     6241 tacgataacc gcgcacgagg atcgtcacat ccgcttctgg gacaacacat ccggcacgct
     6301 ggtacactcc atggtggcgc atttagagcc ggtcacctcg ctggccgtcg atgctcatgg
     6361 cctgtatttg ctatccggct cgcacgactg ctcgatacgg ctgtggaacc tggacaataa
     6421 gacgtgcgtg caggagatca cagcgcatcg caagaagttt gacgagagca tattcgatgt
     6481 ggccttccat gccacaaagc cgtacattgc tagtgccggg gccgatggcc tcgccaaggt
     6541 ctttgtctaa gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct
     6601 aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt
     6661 tatctgacta aatcttagtt tgtattgtca tgttttaata caatatgtta tgtttaaata
     6721 tgtttttaat aaattttata aaataatttc aacttttatt gtaacaacat tgtccattta
     6781 cacactcctt tcaagcgcgt gggatcgatg ctcactcaaa ggcggtaata cggttatcca
     6841 cagaatcagg ggataacgca ggaaagaaca tgtgagccat atgggcccat gtgagccata
     6901 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     6961 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     7021 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     7081 gcga
//
