LOCUS       sequence-463596-        8189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8189)
  AUTHORS   Lopez Martinez D, Todorovski I, Noe Gonzalez M, Rusimbi C, Blears D,
            Khallou N, Han Z, Dirac-Svejstrup AB, Svejstrup JQ
  TITLE     PAF1C-mediated activation of CDK12/13 kinase activity is critical 
            for CTD phosphorylation and transcript elongation.
  JOURNAL   Mol Cell. 2025 May 15;85(10):1952-1967.e8. doi: 
            10.1016/j.molcel.2025.04.012. Epub 2025 May 1.
  PUBMED    40315851
REFERENCE   2  (bases 1 to 8189)
  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"; doi: 
            "10.1016/j.molcel.2025.04.012"; journalName: "Mol Cell"; date: 
            "2025-05-15- 15"; volume: "85"; issue: "10"; pages: "1952-1967.e8"
COMMENT     Sequence Label: pLIB-FLAG(N)-CDK12deltaCTD(1-1082)
FEATURES             Location/Qualifiers
     source          1..8189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(260..848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(340..359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1019..1810,1811..1879))
                     /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     1642..1661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1880..1984)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2010..2464)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2147..2168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2358..2377
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  2648..2813
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    2923..3057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2947..2966)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3001..3020
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6436..6459)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(6466..6489)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(6535..6626)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(6593..6610)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    6674..6707
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(7103..7108,7109..7125,7126..7131)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             7320..7853
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  7920..8144
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
ORIGIN
        1 ttgccaggtt acgcggctgg tctatgcggt gtgaaatacc gcacagatgc gtaaggagaa
       61 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      661 agttcttgaa gtggtggcct aactacggct acactagaag aatagtattt ggtatctgcg
      721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1981 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     2041 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2101 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2161 gagtccacta ttaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2221 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2281 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2341 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2401 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2461 gcgtcccatt cgccattcag gctgcaaata agcgttgata ttcagtcaat tacaaacatt
     2521 aataacgaag agatgacaga aaaattttca ttctgtgaca gagaaaaagt agccgaagat
     2581 gacggtttgt cacatggagt tggcaggatg tttgattaaa aacataacag gaagaaaaat
     2641 gccccgctgt gggcggacaa aatagttggg aactgggagg ggtggaaatg gagtttttaa
     2701 ggattattta gggaagagtg acaaaataga tgggaactgg gtgtagcgtc gtaagctaat
     2761 acgaaaatta aaaatgacaa aatagtttgg aactagattt cacttatctg gttcggatct
     2821 cctaggggtt atgatagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg
     2881 gctttgttag cagccggatc ttctaggctc aagcagtgat cagatccaga catgataaga
     2941 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3001 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3061 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3121 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctagt acttctcgac
     3181 aagcttttag ctgttcttca caggctcagt acttgtccct gaggtagttt cttttcgaga
     3241 agttttggat ggaggtggct cttcgactac aacaccactt tgtcgctgac gtcgccgttt
     3301 cttactccac aactcatggc aatcctgcca gtgggggagg tctggaggag ccattttgct
     3361 gagttcgaca tctttaagga agtcgctctg tagggtctgt tcagctgtgc accgcttact
     3421 aggatctagt gtcagcatgt ggtccaataa atcaagtgct gcagaaggaa tgaaagagaa
     3481 ttcttctcgt agacgccttc gatattgctt cttcggtttc atggtgttga agtagggcag
     3541 tttgataaca tcaggccaca cagctggaca agggctacca caaagtcggc tgatcagttc
     3601 tagctgagcc agttccagat tggcttgaaa aataggcttc tttgtgaata gttccccaag
     3661 aatacatcca cagctccaaa catctatggc tggtgtgtaa cgttcctctc ctagcagtag
     3721 ttctggaggt cggtaccaca aagtaatgac tttgtttgtg taagggcgac tctcttcaga
     3781 gttatagagc cgagcaagtc caaaatctgc tagtttgatt tgcccactgt tattcagcaa
     3841 aatgttagaa cacttaatat cccgatgcag gaaattcttt ttgtgacagt attccaatcc
     3901 ttccattagc tgtttcatga acgacttgat atggtcctca gaaaagtgca ccaaaccaga
     3961 ttctagcagt cccattaagt catggtccat atactcaaat acaaggtaaa aggcaccttt
     4021 gtccttcttg aaatccagtg catcttgttt atctgtgaca atttccttca tgttaacaac
     4081 acttcggtgg attaactgac gaaggatttt gatttcacga atggctgtga ttgggaagcc
     4141 ctctttctca ttgtctagtc tcaccttctt cagagccact agttctcctg tgtctttgtc
     4201 cttggcttta tatacttggc cataggttcc ttctccaata atcccaataa tgtcaaactt
     4261 gtccacacag cgtttccccc agtcgctttc tgtttgtctt ctttctccat aacgaggaca
     4321 acaaattttt ggtctctttt tatatggttg ctgaggtggt gtgattgcct ttggttctgg
     4381 agagtctggg ggagacagat ctccaccagg gagctctgga gggagaggaa ggtctgtgag
     4441 taagtgacgt gtcctctgtt ccttctcttt cttcacaggt tttgaaggaa gagtttcttt
     4501 tggactatcc atgtcatcat ctccaggtaa taagggtggg aggggcaaag gaggcaacgt
     4561 tgaagttttc aggtgtggaa tagcagctgt tacagatact tgagtcttca cagaaacctg
     4621 tacagggggc tgagaatttg cctgagagga cacagcagat gtctttgagt gagtagaagg
     4681 gggcaaagtt gaagtactgg aagcaggaac ctgactaaat gctggctgag aaggaggcag
     4741 aggtggttgc tgtggaagag ctggtattgg aggcaaaggt ggcaaagggg gtgtctgagg
     4801 tggaggggta gtagttggta gagggggtgg gggagaagca attgtgggaa gaggtggagg
     4861 ggtctccttt tctgatgttt ctgtctcctt tggagtaaca atctcctctt tcagtgctat
     4921 gggtttagag tctcttgttc cctgtgcttt caaatcttta acaagatgct tctcggagtt
     4981 ctcatccaac tttactttcc ctgtatctga agaatttttt acctctgtgt ttagatgtgt
     5041 tacattcacc agttcagtat ctggggcaga tttttccaat tttacacttc tgggtaactt
     5101 tttagactcc aaacctgaat ccttagcctc tactgaactg ttctcttttc taggcaaaaa
     5161 tacaggtgaa cccttggact cctttccatc catctttgct gcagcagcag cagctgctct
     5221 ttccttcttt ttcctactga gttcagctcc cagactggaa ttaagtggaa gcctgacagg
     5281 tgagatactg gaatgacgac tgcgtgaact ggatctcttc tttttactat gagaagatga
     5341 atgtcttgaa tatgcaggac ttctacttct ggacttcatg gatttcctac tggggagtgg
     5401 actccgactc agagaccgct tgctcaggaa agggctgctg gaccgccttc gaccataggg
     5461 actgggcgat cgcccgctgt aagagccact tctttcgtag ctggacgacc gtctcctgct
     5521 atagggactg acagatctct gtcgcctact gtaggggctc ggtgaccggg tgctggactg
     5581 gtaggccgaa ggctccttgt aagggggact gaccgactgc cttctcgagg tacttccagg
     5641 acttttcttg taggagtcat aattgctcga ggtatgagat cgtgagggac taaggtcata
     5701 atcttggcca taagaagctc ccgaggggct atcatcttgt ttggagctgt cagaccactt
     5761 cctgtggggg ctcctggatc tccgttttgg gctgtccact gttttgtaac ttttgggtgt
     5821 ttcccttttt cgatgacttt tactccggtc tttgtgccca gacttcagct cccgttcttt
     5881 cctggtcttc tccttgtgga gcttggatga cctggattcc ttgctgctgc ttttggctac
     5941 ctgcgccttc ccatagtcat cagtctcctc attcgaacgc tttgaacttc ccgatatccg
     6001 gtccttcatc gatcccgact tgctggagac ttcttggctt ttttcttttt cggtctgttt
     6061 agcttttagt aagtcccggg aacgcctgtg ctggtggtga cgatgtttgt gcaggcggtc
     6121 gctccgatct gatccacgac gttcgtcgtt ctcccttcgg tctagtttga aggccatgtc
     6181 atcggagaag gtgtcggaat cagagctgat atcatcatac tccaccaaag gtttgataac
     6241 tgtgcccagg gatgctgctt cgggggtcac caaccccatg tctttggagt gtttggactt
     6301 atgccgcttg tgcttcgata ccaagcggtg acgctctctg ctgttagagc tgccgcctcc
     6361 cgatgacggc tgcaaagttc cagaagctcc tccactcccg tccttcttgc ccccatgtct
     6421 ctctgaattg ggcatgggtc cctgaaagag gacttcaaga tcgagcttgt catcgtcgtc
     6481 cttgtagtcc atggatccgc gcccgatggt gggacggtat gaataatccg gaatatttat
     6541 aggttttttt attacaaaac tgttacgaaa acagtaaaat acttatttat ttgcgagatg
     6601 gttatcattt taattatctc catgatctat taatattccg gagtaggtcg cgaatcgata
     6661 ctagtaaaac cagataactt cgtataatgt atgctatacg aagttatggt accgcggccg
     6721 cgtagaggat ctgttgatca gcagttcaac ctgttgatag tacaaacaaa gctgtagcca
     6781 accactagaa ctatagctag agtcctgggc gaacaaacga tgctcgcctt ccagaaaacc
     6841 gaggatgcga accacttcat ccggggtcag caccaccggc aagcgccgcg acggccgagg
     6901 tctaccgatc tcctgaagca agggcagatc cgtgcacagc accttgccgt agaagaacag
     6961 caaggccgcc aatgcctgac gatgcgtgga gaccgaaacc ttgcgctcgt tcgccagcca
     7021 ggacagaaat gcctcgactt cgctgctgcc caaggttgcc gggtgacgca caccgtggaa
     7081 acggatgaag gcacaaaccc agttgacata agcctgttcg gttcgtaaac tgtaatgcaa
     7141 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     7201 cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta tgcctcgggc
     7261 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     7321 tgttacgcag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaggtg
     7381 gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc aaatccatgc
     7441 gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac tcccaacatc
     7501 agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc gcgcttgctg
     7561 ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc aagtttgagc
     7621 agccgcgtag tgagatctat atctatgatc tcgcagtctc cggcgagcac cggaggcagg
     7681 gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt ggtgcttatg
     7741 tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat acaaagttgg
     7801 gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc taacaattcg
     7861 ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca caacgccgcg
     7921 aatatagtct ttaccatgcc cttggccacg cccctcttta atacgacggg caatttgcac
     7981 ttcagaaaat gaagagtttg ctttagccat aacaaaagtc cagtatgctt tttcacagca
     8041 taactggact gatttcagtt tacaactatt ctgtctagtt taagacttta ttgtcatagt
     8101 ttagatctat tttgttcagt ttaagacttt attgtccgcc cacacccgct tacgcagggc
     8161 atccatttat tactcaaccg taaccgatt
//
