LOCUS       sequence-461153-        5801 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 5801)
  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 5801)
  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-WDR61
FEATURES             Location/Qualifiers
     source          1..5801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(715..806)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(773..790)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    854..887
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(1283..1288,1289..1305,1306..1311)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1500..2033
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  2100..2324
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(2458..2475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2629..3217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2709..2728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3388..4179,4180..4248))
                     /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     4011..4030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4249..4353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4379..4833)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4516..4537)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4727..4746
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  5017..5182
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    5292..5426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5316..5335)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5370..5389
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 caaattggca tgttgtacat catagatctt gatgtagcca tcatctgaag cagtgacaag
       61 gagctgggag tccggggaaa aggtcaagga gcgaatgggc atggcatggc cttccagggt
      121 atgcagaagt tttccagttg caatatcaaa aatattgatg attccatcta tggctccact
      181 ggctaggtat ttcccatcag gactatatgc aatactaaga atgaattttc ctctcgtgtc
      241 caaagaatat tcctttttcc cactttccac accaaaaatg ttcactttcc cgacatgagt
      301 tcctgtggcc agatactggg aatcaggaga aaaggccaaa gtccaggcat ccacaggtcc
      361 tgcatctatg gactttatct gtttgccatt ttccaagtcc caaagacgaa tatgagcatc
      421 aagagagctg gatgcagcaa tgggcagggt gtggctgatg tccacagaca ccactcccag
      481 ctgatgtccc tccagactcc actgtaggtc cagcctctca tcacgccatt tccagacctt
      541 caccaggtca tctagggagc ctgtgaccac tgtctcagag ttttccttct tgtttgtccc
      601 ccaagcaact gaccaaatgg catcatcatg ggcttgctct tgtttgaaga gaataccgta
      661 ctggttggtc atggatccgc gcccgatggt gggacggtat gaataatccg gaatatttat
      721 aggttttttt attacaaaac tgttacgaaa acagtaaaat acttatttat ttgcgagatg
      781 gttatcattt taattatctc catgatctat taatattccg gagtaggtcg cgaatcgata
      841 ctagtaaaac cagataactt cgtataatgt atgctatacg aagttatggt accgcggccg
      901 cgtagaggat ctgttgatca gcagttcaac ctgttgatag tacaaacaaa gctgtagcca
      961 accactagaa ctatagctag agtcctgggc gaacaaacga tgctcgcctt ccagaaaacc
     1021 gaggatgcga accacttcat ccggggtcag caccaccggc aagcgccgcg acggccgagg
     1081 tctaccgatc tcctgaagca agggcagatc cgtgcacagc accttgccgt agaagaacag
     1141 caaggccgcc aatgcctgac gatgcgtgga gaccgaaacc ttgcgctcgt tcgccagcca
     1201 ggacagaaat gcctcgactt cgctgctgcc caaggttgcc gggtgacgca caccgtggaa
     1261 acggatgaag gcacaaaccc agttgacata agcctgttcg gttcgtaaac tgtaatgcaa
     1321 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     1381 cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta tgcctcgggc
     1441 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     1501 tgttacgcag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaggtg
     1561 gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc aaatccatgc
     1621 gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac tcccaacatc
     1681 agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc gcgcttgctg
     1741 ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc aagtttgagc
     1801 agccgcgtag tgagatctat atctatgatc tcgcagtctc cggcgagcac cggaggcagg
     1861 gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt ggtgcttatg
     1921 tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat acaaagttgg
     1981 gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc taacaattcg
     2041 ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca caacgccgcg
     2101 aatatagtct ttaccatgcc cttggccacg cccctcttta atacgacggg caatttgcac
     2161 ttcagaaaat gaagagtttg ctttagccat aacaaaagtc cagtatgctt tttcacagca
     2221 taactggact gatttcagtt tacaactatt ctgtctagtt taagacttta ttgtcatagt
     2281 ttagatctat tttgttcagt ttaagacttt attgtccgcc cacacccgct tacgcagggc
     2341 atccatttat tactcaaccg taaccgattt tgccaggtta cgcggctggt ctatgcggtg
     2401 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     2461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3061 cactagaaga atagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4381 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4441 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4501 gggttgagtg ttgttccagt ttggaacaag agtccactat taagaacgtg gactccaacg
     4561 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4621 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4681 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4741 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4801 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcaaataa
     4861 gcgttgatat tcagtcaatt acaaacatta ataacgaaga gatgacagaa aaattttcat
     4921 tctgtgacag agaaaaagta gccgaagatg acggtttgtc acatggagtt ggcaggatgt
     4981 ttgattaaaa acataacagg aagaaaaatg ccccgctgtg ggcggacaaa atagttggga
     5041 actgggaggg gtggaaatgg agtttttaag gattatttag ggaagagtga caaaatagat
     5101 gggaactggg tgtagcgtcg taagctaata cgaaaattaa aaatgacaaa atagtttgga
     5161 actagatttc acttatctgg ttcggatctc ctaggggtta tgatagttat tgctcagcgg
     5221 tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatct tctaggctca
     5281 agcagtgatc agatccagac atgataagat acattgatga gtttggacaa accacaacta
     5341 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     5401 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     5461 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     5521 ctgattatga tcctctagta cttctcgaca agcttttaaa ttggacaatc atagatgtga
     5581 atttcctggt catctccaac agacacaatt tttgaaccat ttccattgta ttttactccc
     5641 cagacctgat cctggtgatc aaagaaggtg tgaacacaag tcctcgttcc aacatcccaa
     5701 acttttacac ttttgtcaga cgaactggaa acaaagtgag tgtcatcagg acagaatgca
     5761 acgttcagca cccaggaggc atggccgctc agcgtgccag c
//
