LOCUS       sequence-461142-        7064 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 7064)
  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 7064)
  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-Cyclin T1
FEATURES             Location/Qualifiers
     source          1..7064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1573..1664)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1631..1648)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    1712..1745
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(2141..2146,2147..2163,2164..2169)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2358..2891
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  2958..3182
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(3316..3333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3487..4075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3567..3586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4246..5037,5038..5106))
                     /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     4869..4888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5107..5211)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5237..5691)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5374..5395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5585..5604
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  5875..6040
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    6150..6284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6174..6193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6228..6247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gtgcttttct ttgtgctctc ggctctttgt aacactgtcc tctacagaat tgtgcttatc
       61 agctgcagca tggactttta tgcgcatttt tatctcctct ggttttgaag acgcagcttt
      121 atctccacct gccactggga ttctcatttt gagagctgtt ttgtcagcct tttccagaaa
      181 aggccgctcg gggttttctg aaccctctat gggcattttt agaatgactg aagaatggct
      241 atcatgatga gaaaggagat tctgggcagc atatgcatat tgtgacttca cattggcttc
      301 catgttctcc agttgcctct tctgggcagc caattcttct gcatgcttcg cgcggtattc
      361 tttcagtgac actttagctg atggcacact cttactattc tgcttctggg aaataaatgc
      421 atttgaacca tcctgtggta aggaatgatc aactcctgta agtgctaaat tctcactagt
      481 ccgatgaccc tgagtaggtt ctagtttgaa agaaggttgg gaggacagcc aacgcttgcc
      541 cggcaacatc tccacactgg ttaagttgct ggatgactct tcggagactg gcagggaagg
      601 cactgcactt gtggtagaag ttgacatgct cattaaacct gcaatggttg tgtctgaaga
      661 gctctgggaa atcatattga ggattgtctg ctctgaagtc ttttcatctg ttcctcggtc
      721 atctgctttt gttttcttgg cagcctcgca tgccctccaa ttccaaatgc gtttgagcct
      781 gttgggagtt ttctccaaaa tctgtagaaa ctcatgtgtc agttcatcta aaagttccaa
      841 ggtcacagtg gcgtcaacat actcccacca gtgcttcccg tcagttgaga ctgggatctc
      901 ccaattggac cacttgcaag ccaggtgaat gcagacacag gccaccacag gaggtgtgta
      961 ctgcaggcta aatgtggtca aatgcaggct gttggttgcc atgaagtaag aagtctgtgc
     1021 taagtccttg cttgctcgaa caagttgagt gcactttact acatgagtat gtgggtgatc
     1081 aattgttagt tcaaagccta aagtctgcaa aattatgctt tctaaaatga ccagatcttg
     1141 aacttgttgc aaataagcct cacttctagt atcaggaagg gattcctgag gatggagaca
     1201 agtatgtgct accttgatga catgttccaa ttttttgggc tgctcctcca ctttagctgc
     1261 tagaaacaag gctgctggag ccacagaatt tccagggaac tgtgtgaagg actgaatcat
     1321 gtagaatcga tgcatgtata ctatagcagt gttgatagtc aattgtgaga cgttaagacg
     1381 ctgccccatg tcctgaagca gattggccgc ctgctggcga taagaaagtt ctttatctgg
     1441 gtccacgcca aaacgacggg atgggctatt ttccagctgt tctcgagtga aataccaccg
     1501 tttgttgttg ttcttcctct ctccctccat ggatccgcgc ccgatggtgg gacggtatga
     1561 ataatccgga atatttatag gtttttttat tacaaaactg ttacgaaaac agtaaaatac
     1621 ttatttattt gcgagatggt tatcatttta attatctcca tgatctatta atattccgga
     1681 gtaggtcgcg aatcgatact agtaaaacca gataacttcg tataatgtat gctatacgaa
     1741 gttatggtac cgcggccgcg tagaggatct gttgatcagc agttcaacct gttgatagta
     1801 caaacaaagc tgtagccaac cactagaact atagctagag tcctgggcga acaaacgatg
     1861 ctcgccttcc agaaaaccga ggatgcgaac cacttcatcc ggggtcagca ccaccggcaa
     1921 gcgccgcgac ggccgaggtc taccgatctc ctgaagcaag ggcagatccg tgcacagcac
     1981 cttgccgtag aagaacagca aggccgccaa tgcctgacga tgcgtggaga ccgaaacctt
     2041 gcgctcgttc gccagccagg acagaaatgc ctcgacttcg ctgctgccca aggttgccgg
     2101 gtgacgcaca ccgtggaaac ggatgaaggc acaaacccag ttgacataag cctgttcggt
     2161 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     2221 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt
     2281 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     2341 gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc
     2401 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
     2461 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag
     2521 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
     2581 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
     2641 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
     2701 gcgagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
     2761 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
     2821 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
     2881 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
     2941 aattgtcaca acgccgcgaa tatagtcttt accatgccct tggccacgcc cctctttaat
     3001 acgacgggca atttgcactt cagaaaatga agagtttgct ttagccataa caaaagtcca
     3061 gtatgctttt tcacagcata actggactga tttcagttta caactattct gtctagttta
     3121 agactttatt gtcatagttt agatctattt tgttcagttt aagactttat tgtccgccca
     3181 cacccgctta cgcagggcat ccatttatta ctcaaccgta accgattttg ccaggttacg
     3241 cggctggtct atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3901 gtggcctaac tacggctaca ctagaagaat agtatttggt atctgcgctc tgctgaagcc
     3961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4201 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4261 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4321 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4381 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4441 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4501 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4561 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4621 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4681 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4741 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4801 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4861 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4921 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4981 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5041 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5101 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5161 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5221 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5281 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5341 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5401 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5461 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5521 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5581 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5641 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     5701 cattcaggct gcaaataagc gttgatattc agtcaattac aaacattaat aacgaagaga
     5761 tgacagaaaa attttcattc tgtgacagag aaaaagtagc cgaagatgac ggtttgtcac
     5821 atggagttgg caggatgttt gattaaaaac ataacaggaa gaaaaatgcc ccgctgtggg
     5881 cggacaaaat agttgggaac tgggaggggt ggaaatggag tttttaagga ttatttaggg
     5941 aagagtgaca aaatagatgg gaactgggtg tagcgtcgta agctaatacg aaaattaaaa
     6001 atgacaaaat agtttggaac tagatttcac ttatctggtt cggatctcct aggggttatg
     6061 atagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
     6121 ccggatcttc taggctcaag cagtgatcag atccagacat gataagatac attgatgagt
     6181 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     6241 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     6301 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     6361 tctacaaatg tggtatggct gattatgatc ctctagtact tctcgacaag cttttactta
     6421 ggaaggggtg gaagtggtgg aggaggttct gatggcagag gtggtggccg gggtttgtct
     6481 gtattgccag atctcgagga gattccacca gaccgaggat tcagatagtc actataagga
     6541 cgaacaaatt caaatgcagt gggctgagtg ggctgaacac cctgggcact gagcagggag
     6601 tgaagcatat tcacagtgtc ttgatagtct attgtctggg tcgtgttgtg accattggcc
     6661 ccagtgggcc ctttatccag tttcgaatga gggacacgag ttttacagct gggctgtgaa
     6721 aaggaaaggc cacttgtgtc tgagctatgc ccaggcatct gacccattgt aggaagtgaa
     6781 gggaaggaga aatttaggga agaggattta gtactcttgg caatcttggc tggatgatca
     6841 aacacagccc ctccagtctc ttcagagggt ccccttttac gagtagaact ggaagaggaa
     6901 aaagaactag acaagctata ggttttatgt gctaagttgc ttgtctggct actatgtttt
     6961 ggatcaccag gacgtttgtt cccagtacca actggaagtt gggaatgaga gtgcttgtgt
     7021 gagtggtgat tatgatgatg atgatgatta gatgggtgag tctt
//
