LOCUS       sequence-461134-        6431 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 6431)
  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 6431)
  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)-CDC73deltaKIM(delta290-324)
FEATURES             Location/Qualifiers
     source          1..6431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1528..1551)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(1558..1581)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1627..1718)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1685..1702)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    1766..1799
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(2195..2200,2201..2217,2218..2223)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2412..2945
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  3012..3236
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(3370..3387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3541..4129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3621..3640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4300..5091,5092..5160))
                     /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     4923..4942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5161..5265)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5291..5745)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5428..5449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5639..5658
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  5929..6094
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    6204..6338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6228..6247)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6282..6301
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gctgattatg atcctctagt acttctcgac aagctttcag aatctcaagt gcgatttatg
       61 ctttaccatg tacctgtcca atgtttccca aaaccgtaag aacactggtc tatccaaatg
      121 acgtttgtga tagctgagtt ctaatactgt tacatcccat ttctgaacat ttggatccag
      181 acgaacttca tcatacttca gatggaaggc tttaatttta gcaaatatat caactggtga
      241 tccatcaggc aaaagccatg gccaaccttt gaactgccat gcaggaccct gcacaaaaac
      301 ggctacaacg cggtcccagt cttgaggcat aagtttaagg ggctggtcta ctactctata
      361 aggtactgta acactaattg cagtgccccc tggttgcatc tggtcttttc ttctttgtat
      421 tagagtttca ttttctcgtt gacaaccttg tttcttcttt tcatctgatg ggacaaattt
      481 caggtcctgt agaaggtctt ttgcattaag catggttatt aaagaggtgg tagctgcagg
      541 aattatgata atgggtgttc gagatccttt cttctgattt gggggaggtc ttgcttgaga
      601 aactggtctt ggtactggct gggctgcagg agtctgagtc ttccgggcag atgcaccctc
      661 ggcagctggg ataggctgtt tggtgcgcaa agtgggatcc acaggtgctg catttggggc
      721 aggtcgctgt tcaggtgcac gcccttcttc tctggctttt acagattgaa gaattgcaaa
      781 aatgttcttg gaaaaattct ttcctgtgct ttgtaagata gttgttcgtg tcctccatac
      841 tctctctctg ctgacaatat ctcgggtcac atctacctca gcatccacaa aactcctctg
      901 tttaagggca gttatgtcat catctagatc agtcttgata gtagatcttt tcttagccat
      961 aattttggct ttgattgcag caattttttc cactgacata gcttcagaca aagacctaat
     1021 ctgttcagtc tgtacaatcc cttctttgtg accctccaaa cgggcagcca atctctcttt
     1081 atcaaggcgc acacactctt catcctcaat tcgtggtttc tttgcttctg ctaaaacttc
     1141 atctgcagct cgtttgactt gagtagatcg ctgaagacct atttctaagg gagcgcttct
     1201 gtctatactt gccgatgttg acgcttcacc attgagatat ccaagtagat cttttcgatc
     1261 aggtcttcta accacaggaa tattttcagt agctgcacgt cggacataaa caggatgaga
     1321 aaggtgcacg ttattaagta gaaataaaat ggaatccaat gtgtagtact ctctgggttg
     1381 gccttccttt ccagtccccc aaacaacata gttggtcttc acattcttgg gccaggagaa
     1441 ctccccgaag atcacttcgt ctcccttcac cacaatctcc ttcttctgga tgttgtactg
     1501 tcgcaggacg ctaagcacgt ccgccatggg tccctgaaag aggacttcaa gatcgagctt
     1561 gtcatcgtcg tccttgtagt ccatggatcc gcgcccgatg gtgggacggt atgaataatc
     1621 cggaatattt ataggttttt ttattacaaa actgttacga aaacagtaaa atacttattt
     1681 atttgcgaga tggttatcat tttaattatc tccatgatct attaatattc cggagtaggt
     1741 cgcgaatcga tactagtaaa accagataac ttcgtataat gtatgctata cgaagttatg
     1801 gtaccgcggc cgcgtagagg atctgttgat cagcagttca acctgttgat agtacaaaca
     1861 aagctgtagc caaccactag aactatagct agagtcctgg gcgaacaaac gatgctcgcc
     1921 ttccagaaaa ccgaggatgc gaaccacttc atccggggtc agcaccaccg gcaagcgccg
     1981 cgacggccga ggtctaccga tctcctgaag caagggcaga tccgtgcaca gcaccttgcc
     2041 gtagaagaac agcaaggccg ccaatgcctg acgatgcgtg gagaccgaaa ccttgcgctc
     2101 gttcgccagc caggacagaa atgcctcgac ttcgctgctg cccaaggttg ccgggtgacg
     2161 cacaccgtgg aaacggatga aggcacaaac ccagttgaca taagcctgtt cggttcgtaa
     2221 actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     2281 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc
     2341 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2401 gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     2461 caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag
     2521 tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct
     2581 actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca
     2641 tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc
     2701 ccaagtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc
     2761 accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc
     2821 ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct
     2881 atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca
     2941 cctaacaatt cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt
     3001 cacaacgccg cgaatatagt ctttaccatg cccttggcca cgcccctctt taatacgacg
     3061 ggcaatttgc acttcagaaa atgaagagtt tgctttagcc ataacaaaag tccagtatgc
     3121 tttttcacag cataactgga ctgatttcag tttacaacta ttctgtctag tttaagactt
     3181 tattgtcata gtttagatct attttgttca gtttaagact ttattgtccg cccacacccg
     3241 cttacgcagg gcatccattt attactcaac cgtaaccgat tttgccaggt tacgcggctg
     3301 gtctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     3361 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3421 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3961 taactacggc tacactagaa gaatagtatt tggtatctgc gctctgctga agccagttac
     4021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4081 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4141 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4201 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4261 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4321 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4381 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4441 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4501 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4561 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4621 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4681 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4741 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4801 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4861 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4921 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4981 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5041 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5101 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5161 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5221 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5281 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5341 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5401 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaagaacg
     5461 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5521 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5581 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5641 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5701 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     5761 ggctgcaaat aagcgttgat attcagtcaa ttacaaacat taataacgaa gagatgacag
     5821 aaaaattttc attctgtgac agagaaaaag tagccgaaga tgacggtttg tcacatggag
     5881 ttggcaggat gtttgattaa aaacataaca ggaagaaaaa tgccccgctg tgggcggaca
     5941 aaatagttgg gaactgggag gggtggaaat ggagttttta aggattattt agggaagagt
     6001 gacaaaatag atgggaactg ggtgtagcgt cgtaagctaa tacgaaaatt aaaaatgaca
     6061 aaatagtttg gaactagatt tcacttatct ggttcggatc tcctaggggt tatgatagtt
     6121 attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat
     6181 cttctaggct caagcagtga tcagatccag acatgataag atacattgat gagtttggac
     6241 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     6301 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     6361 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     6421 aatgtggtat g
//
