LOCUS       sequence-461148-        6428 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 6428)
  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 6428)
  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-GST(N)-CDK12RS(104-380)
FEATURES             Location/Qualifiers
     source          1..6428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(86..109)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(110..763)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(130..152)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(770..793)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(839..930)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(897..914)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    978..1011
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(1407..1412,1413..1429,1430..1435)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1624..2157
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  2224..2448
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(2582..2599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2753..3341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2833..2852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3512..4303,4304..4372))
                     /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     4135..4154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4373..4477)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4503..4957)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4640..4661)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4851..4870
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  5141..5306
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    5416..5550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5440..5459)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5494..5513
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tttctttttc ggtctgttta gcttttagta agtcccggga acgcctgtgc tggtggtgac
       61 gatgtttgtg caggcggtcg ctccggggtc cctgaaagag gacttcaagt tttggaggat
      121 ggtcgccacc accaaacgtg gcttgccagc cctgcaaagg ccatgctata tacttgctgg
      181 atttcaagta cttatcaatt tgtgggatag cttcaatacg ttttttaaaa caaactaatt
      241 ttgggaacgc atccaggcac attgggtcca tgtataaaac aacatcaaga gcgtcataca
      301 acatgaagtc aggatgggtt acatgatcac catttaaata tgttttatga cataaacgat
      361 cttcgaacat tttcagcatt tcaggtagct tgctaagaaa atcaactttg agagtttcaa
      421 agtctttact atatgcaatt ctcgaaacac cgtatctaat atccaaaacc gctccttcaa
      481 gcattgaaat ctctgcacgc tcttttggac aaccacccaa catgttgtgc ttgtcagcta
      541 tataacgtat gatggccata gactgtgtta atttaacatc accatcaata taataaggaa
      601 gattgggaaa ctccaaaccc aattcaaact ttttgtttcg ccatttatca ccttcatcgc
      661 gctcatacaa atgctcttca tatttttctt caagatattc caaaagaagt cgagtgggtt
      721 gcacaaggcc cttaattttc caataaccta gtatagggga catatcgagc ttgtcatcgt
      781 cgtccttgta gtccatggat ccgcgcccga tggtgggacg gtatgaataa tccggaatat
      841 ttataggttt ttttattaca aaactgttac gaaaacagta aaatacttat ttatttgcga
      901 gatggttatc attttaatta tctccatgat ctattaatat tccggagtag gtcgcgaatc
      961 gatactagta aaaccagata acttcgtata atgtatgcta tacgaagtta tggtaccgcg
     1021 gccgcgtaga ggatctgttg atcagcagtt caacctgttg atagtacaaa caaagctgta
     1081 gccaaccact agaactatag ctagagtcct gggcgaacaa acgatgctcg ccttccagaa
     1141 aaccgaggat gcgaaccact tcatccgggg tcagcaccac cggcaagcgc cgcgacggcc
     1201 gaggtctacc gatctcctga agcaagggca gatccgtgca cagcaccttg ccgtagaaga
     1261 acagcaaggc cgccaatgcc tgacgatgcg tggagaccga aaccttgcgc tcgttcgcca
     1321 gccaggacag aaatgcctcg acttcgctgc tgcccaaggt tgccgggtga cgcacaccgt
     1381 ggaaacggat gaaggcacaa acccagttga cataagcctg ttcggttcgt aaactgtaat
     1441 gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
     1501 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag tctatgcctc
     1561 gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca
     1621 acgatgttac gcagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
     1681 ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca agtcaaatcc
     1741 atgcgggctg ctcttgatct tttcggtcgt gagttcggag acgtagccac ctactcccaa
     1801 catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt catcgcgctt
     1861 gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct gcccaagttt
     1921 gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggcga gcaccggagg
     1981 cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc gcttggtgct
     2041 tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct ctatacaaag
     2101 ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc cacctaacaa
     2161 ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt tcggtaaatt gtcacaacgc
     2221 cgcgaatata gtctttacca tgcccttggc cacgcccctc tttaatacga cgggcaattt
     2281 gcacttcaga aaatgaagag tttgctttag ccataacaaa agtccagtat gctttttcac
     2341 agcataactg gactgatttc agtttacaac tattctgtct agtttaagac tttattgtca
     2401 tagtttagat ctattttgtt cagtttaaga ctttattgtc cgcccacacc cgcttacgca
     2461 gggcatccat ttattactca accgtaaccg attttgccag gttacgcggc tggtctatgc
     2521 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     2581 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2641 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2701 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2761 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2821 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2881 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2941 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3001 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3061 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3121 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3181 gctacactag aagaatagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3241 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3301 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3361 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3421 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3481 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3541 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3601 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3661 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3721 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3781 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3841 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3901 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3961 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4021 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4081 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4141 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4201 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4261 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4321 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4381 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4441 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4501 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4561 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4621 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaagaa cgtggactcc
     4681 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4741 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4801 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4861 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4921 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcaa
     4981 ataagcgttg atattcagtc aattacaaac attaataacg aagagatgac agaaaaattt
     5041 tcattctgtg acagagaaaa agtagccgaa gatgacggtt tgtcacatgg agttggcagg
     5101 atgtttgatt aaaaacataa caggaagaaa aatgccccgc tgtgggcgga caaaatagtt
     5161 gggaactggg aggggtggaa atggagtttt taaggattat ttagggaaga gtgacaaaat
     5221 agatgggaac tgggtgtagc gtcgtaagct aatacgaaaa ttaaaaatga caaaatagtt
     5281 tggaactaga tttcacttat ctggttcgga tctcctaggg gttatgatag ttattgctca
     5341 gcggtggcag cagccaactc agcttccttt cgggctttgt tagcagccgg atcttctagg
     5401 ctcaagcagt gatcagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     5461 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     5521 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     5581 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     5641 atggctgatt atgatcctct agtacttctc gacaagcttt caacgactgc gtgaactgga
     5701 tctcttcttt ttactatgag aagatgaatg tcttgaatat gcaggacttc tacttctgga
     5761 cttcatggat ttcctactgg ggagtggact ccgactcaga gaccgcttgc tcaggaaagg
     5821 gctgctggac cgccttcgac catagggact gggcgatcgc ccgctgtaag agccacttct
     5881 ttcgtagctg gacgaccgtc tcctgctata gggactgaca gatctctgtc gcctactgta
     5941 ggggctcggt gaccgggtgc tggactggta ggccgaaggc tccttgtaag ggggactgac
     6001 cgactgcctt ctcgaggtac ttccaggact tttcttgtag gagtcataat tgctcgaggt
     6061 atgagatcgt gagggactaa ggtcataatc ttggccataa gaagctcccg aggggctatc
     6121 atcttgtttg gagctgtcag accacttcct gtgggggctc ctggatctcc gttttgggct
     6181 gtccactgtt ttgtaacttt tgggtgtttc cctttttcga tgacttttac tccggtcttt
     6241 gtgcccagac ttcagctccc gttctttcct ggtcttctcc ttgtggagct tggatgacct
     6301 ggattccttg ctgctgcttt tggctacctg cgccttccca tagtcatcag tctcctcatt
     6361 cgaacgcttt gaacttcccg atatccggtc cttcatcgat cccgacttgc tggagacttc
     6421 ttggcttt
//
