LOCUS       sequence-461159-        6056 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 6056)
  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 6056)
  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)-CDK9
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(94..111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(265..853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(345..364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1024..1815,1816..1884))
                     /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     1647..1666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1885..1989)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2015..2469)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2152..2173)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2363..2382
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  2653..2818
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    2928..3062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2952..2971)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3006..3025
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4308..4331)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(4338..4361)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(4407..4498)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(4465..4482)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    4546..4579
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(4975..4980,4981..4997,4998..5003)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5192..5725
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  5792..6016
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
ORIGIN
        1 cgattttgcc aggttacgcg gctggtctat gcggtgtgaa ataccgcaca gatgcgtaag
       61 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
      121 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      181 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      241 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      301 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      361 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      421 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      481 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      541 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      601 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      661 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaatag tatttggtat
      721 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      781 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      841 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
      901 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
      961 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1021 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1081 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1141 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1201 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1261 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1321 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1381 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1441 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1501 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1561 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1621 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1681 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1741 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1801 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     1861 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1921 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1981 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     2041 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2101 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2161 aacaagagtc cactattaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2221 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2281 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2341 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2401 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2461 agggcgcgtc ccattcgcca ttcaggctgc aaataagcgt tgatattcag tcaattacaa
     2521 acattaataa cgaagagatg acagaaaaat tttcattctg tgacagagaa aaagtagccg
     2581 aagatgacgg tttgtcacat ggagttggca ggatgtttga ttaaaaacat aacaggaaga
     2641 aaaatgcccc gctgtgggcg gacaaaatag ttgggaactg ggaggggtgg aaatggagtt
     2701 tttaaggatt atttagggaa gagtgacaaa atagatggga actgggtgta gcgtcgtaag
     2761 ctaatacgaa aattaaaaat gacaaaatag tttggaacta gatttcactt atctggttcg
     2821 gatctcctag gggttatgat agttattgct cagcggtggc agcagccaac tcagcttcct
     2881 ttcgggcttt gttagcagcc ggatcttcta ggctcaagca gtgatcagat ccagacatga
     2941 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3001 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3061 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3121 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcct ctagtacttc
     3181 tcgacaagct ttcagaagac gcgctcaaac tccgtctggt tggtggtggc gggattgcga
     3241 ctctggttgg tggactgctg ggtgatctgg ctgcccttcc ggcgcggtgg tgccaagtac
     3301 tcgaacatgg acgtcaggtg ggtggagagc atgcccttga ggtcggaggg catggggtcg
     3361 gaccagaaga agtcgtggtt gagggcgtca tcgctgtcga tgcgctgggc agggtccagc
     3421 accagcagct tgtcgatgag gtccagtgcg tatgggtcac gcacataggc cttcagcctg
     3481 tccttcacct tccgcttctg gcccttgacc agctccagct tttcgtacag ctcatagttg
     3541 tccacgtttg gccacacctc aggggtgatg gagccgcaga gctgactgat gagggcgagt
     3601 tggtgctgct ccgtgttgcc ctgcatgatg gggctgcggg tccacatctc tgccatgatg
     3661 cacccagcac cccacaggtc aatggggggg ccgtagtccc gctccccgag caacagctcc
     3721 gggggccggt accagagtgt caccacacgg ttggtgtagc ggttgggctg gctgttcttg
     3781 gccaggctga aggcccgggc cagcccaaag tctgccagct tcaggacccc atcacgagtg
     3841 ataagcacat tagcagcctt catgtcccta tgcaggatct tgtttctgtg gatgtagtag
     3901 aggccgttaa gcagcatctg catcaccctc ttgatctcag acagcgtgaa cttgaccaaa
     3961 acattgctca acagcccagc aaggtcatgc tcgcagaagt cgaacaccag gtatatacta
     4021 cccttgcagc ggttataggg ggaagctttg gttcgacaaa tctcaatcaa gttgaccaca
     4081 ttctcgtgtt ttagaagctg aaggatcttg atctcccgca aggctgtaat ggggaacccc
     4141 tccttctcgt tttccatcag caccttcttc agagccacct tctggccggt cttgcggtgc
     4201 ctggccttga acacctcccc gaaggtgcct tggccgatct tggcgagctt ctcgtatttg
     4261 gaaacttcat cacaaaaagg gcactccacc gagtcgtact gctttgcggg tccctgaaag
     4321 aggacttcaa gatcgagctt gtcatcgtcg tccttgtagt ccatggatcc gcgcccgatg
     4381 gtgggacggt atgaataatc cggaatattt ataggttttt ttattacaaa actgttacga
     4441 aaacagtaaa atacttattt atttgcgaga tggttatcat tttaattatc tccatgatct
     4501 attaatattc cggagtaggt cgcgaatcga tactagtaaa accagataac ttcgtataat
     4561 gtatgctata cgaagttatg gtaccgcggc cgcgtagagg atctgttgat cagcagttca
     4621 acctgttgat agtacaaaca aagctgtagc caaccactag aactatagct agagtcctgg
     4681 gcgaacaaac gatgctcgcc ttccagaaaa ccgaggatgc gaaccacttc atccggggtc
     4741 agcaccaccg gcaagcgccg cgacggccga ggtctaccga tctcctgaag caagggcaga
     4801 tccgtgcaca gcaccttgcc gtagaagaac agcaaggccg ccaatgcctg acgatgcgtg
     4861 gagaccgaaa ccttgcgctc gttcgccagc caggacagaa atgcctcgac ttcgctgctg
     4921 cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacaaac ccagttgaca
     4981 taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc
     5041 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     5101 gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     5161 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca
     5221 gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg
     5281 taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga
     5341 gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt
     5401 gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc
     5461 tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct atatctatga
     5521 tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca tcaatctcct
     5581 caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga
     5641 cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga
     5701 tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccgc
     5761 ggagttgttc ggtaaattgt cacaacgccg cgaatatagt ctttaccatg cccttggcca
     5821 cgcccctctt taatacgacg ggcaatttgc acttcagaaa atgaagagtt tgctttagcc
     5881 ataacaaaag tccagtatgc tttttcacag cataactgga ctgatttcag tttacaacta
     5941 ttctgtctag tttaagactt tattgtcata gtttagatct attttgttca gtttaagact
     6001 ttattgtccg cccacacccg cttacgcagg gcatccattt attactcaac cgtaac
//
