LOCUS       sequence-463311-        9413 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 9413)
  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 9413)
  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)-CDK12
FEATURES             Location/Qualifiers
     source          1..9413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(139..243)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(269..723)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(406..427)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     617..636
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  907..1072
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    1182..1316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1206..1225)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1260..1279
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5919..5942)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(5949..5972)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(6018..6109)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(6076..6093)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    6157..6190
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(6586..6591,6592..6608,6609..6614)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6803..7336
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  7403..7627
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(7761..7778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7932..8520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8012..8031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8691..9413,1..69,70..138))
                     /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     9314..9333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
       61 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      241 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
      301 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
      361 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
      421 agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
      481 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
      541 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
      601 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
      661 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
      721 cgtcccattc gccattcagg ctgcaaataa gcgttgatat tcagtcaatt acaaacatta
      781 ataacgaaga gatgacagaa aaattttcat tctgtgacag agaaaaagta gccgaagatg
      841 acggtttgtc acatggagtt ggcaggatgt ttgattaaaa acataacagg aagaaaaatg
      901 ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg agtttttaag
      961 gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg taagctaata
     1021 cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg ttcggatctc
     1081 ctaggggtta tgatagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg
     1141 ctttgttagc agccggatct tctaggctca agcagtgatc agatccagac atgataagat
     1201 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1261 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1321 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     1381 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagta cttctcgaca
     1441 agcttttagt aaggaactcc tctccctctt ccccctcttg gtgggactct tgtaggcccc
     1501 cgatagagtt ttccataagc agaagactga gttgggcccc cccagtgaag gcctgctcct
     1561 gagctgctgg ccccagtggt tcctggcccc agctccccat agttttgcaa tttggtctct
     1621 gcatgtacca cagaattgct gcttccctca gccttcagga atggttgccc gaccaccggg
     1681 tgtaacgcta aggggaccct ggcaaaacgg aggtcctggt cccctggaaa ctgcactgac
     1741 cctgtgccct ggtaactgct ggactcccca aggtggctca gagagcctga gaaggtcctg
     1801 ttcttcacca gggtctggac caaggattct gtcaaggctg gaccagagtt tcgttcatca
     1861 gtgtcaatgg cactgaaccc tgtttcaggc ccatcagtgt ttccataagt cctgtttgga
     1921 cggggtcggg tggagtactc cattggtctc aaggcctggt gctcatgtgg caggtggcca
     1981 ggaggctctg cttcaggctg ggataaaagt tgcagcaagg cggctgtcac ggctgggttc
     2041 aactcctggg gggcgctgga aagatcgcct tcaaccagag ggggtggagg tggcggcggt
     2101 ggaggtccgg ggggctcagg gggcctcttc tctggtggaa gaatgtgagg aggacatgct
     2161 gctgcctcct cctgtggcat tgtgggagtt cttcggggcc cctgtggccc actgttcttg
     2221 tcactgttgt tttcctccag actgcctgct ggctcttggg ttttcatcag ctgactcaag
     2281 agaactgcca atatattctg catgtcagct ggtgtgcttg aagcttcaag ggtcgtctgt
     2341 tctgctgaag gcaggatcac tggggcagag ggtgcttcct tcaaagactc ctctggggcc
     2401 atggtctctg agtcctgctg ctgggaagta gcttccgtca gggcactgat ggattggttc
     2461 agggcttcca gctgctgctg catctctggg ttggagtgga tgttaagcag ctgtgccatt
     2521 tgagggatgc tcaggtcggt ttggctctgc agcaggttta ataacactgc caattcactt
     2581 tgattcagct gttgtgtgat gtcagcaagg cctattgcat ccccagcccc agactccacc
     2641 ttgccaggag caggctgagg tggtgctggg ctgctgttct tcacaggctc agtacttgtc
     2701 cctgaggtag tttcttttcg agaagttttg gatggaggtg gctcttcgac tacaacacca
     2761 ctttgtcgct gacgtcgccg tttcttactc cacaactcat ggcaatcctg ccagtggggg
     2821 aggtctggag gagccatttt gctgagttcg acatctttaa ggaagtcgct ctgtagggtc
     2881 tgttcagctg tgcaccgctt actaggatct agtgtcagca tgtggtccaa taaatcaagt
     2941 gctgcagaag gaatgaaaga gaattcttct cgtagacgcc ttcgatattg cttcttcggt
     3001 ttcatggtgt tgaagtaggg cagtttgata acatcaggcc acacagctgg acaagggcta
     3061 ccacaaagtc ggctgatcag ttctagctga gccagttcca gattggcttg aaaaataggc
     3121 ttctttgtga atagttcccc aagaatacat ccacagctcc aaacatctat ggctggtgtg
     3181 taacgttcct ctcctagcag tagttctgga ggtcggtacc acaaagtaat gactttgttt
     3241 gtgtaagggc gactctcttc agagttatag agccgagcaa gtccaaaatc tgctagtttg
     3301 atttgcccac tgttattcag caaaatgtta gaacacttaa tatcccgatg caggaaattc
     3361 tttttgtgac agtattccaa tccttccatt agctgtttca tgaacgactt gatatggtcc
     3421 tcagaaaagt gcaccaaacc agattctagc agtcccatta agtcatggtc catatactca
     3481 aatacaaggt aaaaggcacc tttgtccttc ttgaaatcca gtgcatcttg tttatctgtg
     3541 acaatttcct tcatgttaac aacacttcgg tggattaact gacgaaggat tttgatttca
     3601 cgaatggctg tgattgggaa gccctctttc tcattgtcta gtctcacctt cttcagagcc
     3661 actagttctc ctgtgtcttt gtccttggct ttatatactt ggccataggt tccttctcca
     3721 ataatcccaa taatgtcaaa cttgtccaca cagcgtttcc cccagtcgct ttctgtttgt
     3781 cttctttctc cataacgagg acaacaaatt tttggtctct ttttatatgg ttgctgaggt
     3841 ggtgtgattg cctttggttc tggagagtct gggggagaca gatctccacc agggagctct
     3901 ggagggagag gaaggtctgt gagtaagtga cgtgtcctct gttccttctc tttcttcaca
     3961 ggttttgaag gaagagtttc ttttggacta tccatgtcat catctccagg taataagggt
     4021 gggaggggca aaggaggcaa cgttgaagtt ttcaggtgtg gaatagcagc tgttacagat
     4081 acttgagtct tcacagaaac ctgtacaggg ggctgagaat ttgcctgaga ggacacagca
     4141 gatgtctttg agtgagtaga agggggcaaa gttgaagtac tggaagcagg aacctgacta
     4201 aatgctggct gagaaggagg cagaggtggt tgctgtggaa gagctggtat tggaggcaaa
     4261 ggtggcaaag ggggtgtctg aggtggaggg gtagtagttg gtagaggggg tgggggagaa
     4321 gcaattgtgg gaagaggtgg aggggtctcc ttttctgatg tttctgtctc ctttggagta
     4381 acaatctcct ctttcagtgc tatgggttta gagtctcttg ttccctgtgc tttcaaatct
     4441 ttaacaagat gcttctcgga gttctcatcc aactttactt tccctgtatc tgaagaattt
     4501 tttacctctg tgtttagatg tgttacattc accagttcag tatctggggc agatttttcc
     4561 aattttacac ttctgggtaa ctttttagac tccaaacctg aatccttagc ctctactgaa
     4621 ctgttctctt ttctaggcaa aaatacaggt gaacccttgg actcctttcc atccatcttt
     4681 gctgcagcag cagcagctgc tctttccttc tttttcctac tgagttcagc tcccagactg
     4741 gaattaagtg gaagcctgac aggtgagata ctggaatgac gactgcgtga actggatctc
     4801 ttctttttac tatgagaaga tgaatgtctt gaatatgcag gacttctact tctggacttc
     4861 atggatttcc tactggggag tggactccga ctcagagacc gcttgctcag gaaagggctg
     4921 ctggaccgcc ttcgaccata gggactgggc gatcgcccgc tgtaagagcc acttctttcg
     4981 tagctggacg accgtctcct gctataggga ctgacagatc tctgtcgcct actgtagggg
     5041 ctcggtgacc gggtgctgga ctggtaggcc gaaggctcct tgtaaggggg actgaccgac
     5101 tgccttctcg aggtacttcc aggacttttc ttgtaggagt cataattgct cgaggtatga
     5161 gatcgtgagg gactaaggtc ataatcttgg ccataagaag ctcccgaggg gctatcatct
     5221 tgtttggagc tgtcagacca cttcctgtgg gggctcctgg atctccgttt tgggctgtcc
     5281 actgttttgt aacttttggg tgtttccctt tttcgatgac ttttactccg gtctttgtgc
     5341 ccagacttca gctcccgttc tttcctggtc ttctccttgt ggagcttgga tgacctggat
     5401 tccttgctgc tgcttttggc tacctgcgcc ttcccatagt catcagtctc ctcattcgaa
     5461 cgctttgaac ttcccgatat ccggtccttc atcgatcccg acttgctgga gacttcttgg
     5521 cttttttctt tttcggtctg tttagctttt agtaagtccc gggaacgcct gtgctggtgg
     5581 tgacgatgtt tgtgcaggcg gtcgctccga tctgatccac gacgttcgtc gttctccctt
     5641 cggtctagtt tgaaggccat gtcatcggag aaggtgtcgg aatcagagct gatatcatca
     5701 tactccacca aaggtttgat aactgtgccc agggatgctg cttcgggggt caccaacccc
     5761 atgtctttgg agtgtttgga cttatgccgc ttgtgcttcg ataccaagcg gtgacgctct
     5821 ctgctgttag agctgccgcc tcccgatgac ggctgcaaag ttccagaagc tcctccactc
     5881 ccgtccttct tgcccccatg tctctctgaa ttgggcatgg gtccctgaaa gaggacttca
     5941 agatcgagct tgtcatcgtc gtccttgtag tccatggatc cgcgcccgat ggtgggacgg
     6001 tatgaataat ccggaatatt tataggtttt tttattacaa aactgttacg aaaacagtaa
     6061 aatacttatt tatttgcgag atggttatca ttttaattat ctccatgatc tattaatatt
     6121 ccggagtagg tcgcgaatcg atactagtaa aaccagataa cttcgtataa tgtatgctat
     6181 acgaagttat ggtaccgcgg ccgcgtagag gatctgttga tcagcagttc aacctgttga
     6241 tagtacaaac aaagctgtag ccaaccacta gaactatagc tagagtcctg ggcgaacaaa
     6301 cgatgctcgc cttccagaaa accgaggatg cgaaccactt catccggggt cagcaccacc
     6361 ggcaagcgcc gcgacggccg aggtctaccg atctcctgaa gcaagggcag atccgtgcac
     6421 agcaccttgc cgtagaagaa cagcaaggcc gccaatgcct gacgatgcgt ggagaccgaa
     6481 accttgcgct cgttcgccag ccaggacaga aatgcctcga cttcgctgct gcccaaggtt
     6541 gccgggtgac gcacaccgtg gaaacggatg aaggcacaaa cccagttgac ataagcctgt
     6601 tcggttcgta aactgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     6661 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     6721 tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     6781 ttgatgttat ggagcagcaa cgatgttacg cagcagcaac gatgttacgc agcagggcag
     6841 tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat gtaggctcgg
     6901 ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga
     6961 cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact tgctccgtag
     7021 taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc
     7081 ttacgttctg cccaagtttg agcagccgcg tagtgagatc tatatctatg atctcgcagt
     7141 ctccggcgag caccggaggc agggcattgc caccgcgctc atcaatctcc tcaagcatga
     7201 ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg acgatcccgc
     7261 agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg atatcgaccc
     7321 aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttcccggccg cggagttgtt
     7381 cggtaaattg tcacaacgcc gcgaatatag tctttaccat gcccttggcc acgcccctct
     7441 ttaatacgac gggcaatttg cacttcagaa aatgaagagt ttgctttagc cataacaaaa
     7501 gtccagtatg ctttttcaca gcataactgg actgatttca gtttacaact attctgtcta
     7561 gtttaagact ttattgtcat agtttagatc tattttgttc agtttaagac tttattgtcc
     7621 gcccacaccc gcttacgcag ggcatccatt tattactcaa ccgtaaccga ttttgccagg
     7681 ttacgcggct ggtctatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     7741 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7801 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7861 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8101 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8341 aagtggtggc ctaactacgg ctacactaga agaatagtat ttggtatctg cgctctgctg
     8401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8701 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8761 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8821 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8881 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8941 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9001 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9061 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9121 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9181 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9241 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9301 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9361 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cag
//
