LOCUS       sequence-461157-        6479 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 6479)
  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 6479)
  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-PAF1
FEATURES             Location/Qualifiers
     source          1..6479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1743..1877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1780..1799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1834..1853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1987..2152
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2336..2790
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2423..2442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2632..2653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2816..2920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2921..2989,2990..3781)
                     /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     complement(3139..3158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3952..4540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4441..4460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4694..4711
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4845..5069
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5136..5669)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5858..5863,5864..5880,5881..5886))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(6282..6315)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6363..6454
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6379..6396
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 ccatcgggcg cggatccatg gcgcccacca tccagaccca ggcccagcgg gaggatggcc
       61 acaggcccaa ttcccaccgg actctgcctg agaggtctgg agtggtctgc cgagtcaagt
      121 actgcaatag cctccctgat atccccttcg accccaagtt catcacctac cccttcgacc
      181 agaacaggtt cgtccagtac aaagccactt ccttggagaa acagcacaaa catgacctcc
      241 tgactgagcc agacctgggg gtcaccatcg atctcatcaa tcctgacacc taccgcatcg
      301 accccaatgt tcttctagat ccagctgatg agaaactttt ggaagaggag attcaggccc
      361 ccaccagctc caagagatcc cagcagcacg cgaaggtggt gccatggatg cgaaagacag
      421 agtacatctc cactgagttc aaccgttatg gcatctccaa tgagaagcct gaggtcaaga
      481 ttggggtttc tgtgaagcag cagtttaccg aggaagaaat atacaaagac agggatagcc
      541 agatcacagc cattgagaag acttttgagg atgcccagaa atcaatctca cagcattaca
      601 gcaaaccccg agtcacaccg gtggaggtca tgcctgtctt cccagacttt aagatgtgga
      661 tcaatccatg tgctcaggtg atctttgact cagacccagc ccccaaggac acgagtggtg
      721 cagctgcgtt ggagatgatg tctcaggcca tgattagggg catgatggat gaggaaggga
      781 accagtttgt ggcctatttc ctgcctgtag aagagacgtt gaagaaacga aagcgggacc
      841 aggaggagga gatggactat gcaccagatg atgtgtatga ctacaaaatt gctcgggagt
      901 acaactggaa cgtgaagaac aaagctagca agggctatga ggaaaactac ttcttcatct
      961 tccgagaggg tgacggggtt tactacaatg agttggaaac cagggtccgc cttagtaagc
     1021 gccgggccaa ggctggggtt cagtcaggca ccaacgccct gcttgtggtc aaacatcggg
     1081 acatgaatga gaaggaactg gaagctcagg aggcacggaa ggcccagcta gaaaaccacg
     1141 aaccggagga ggaagaggaa gaggagatgg agacagaaga gaaagaagct gggggctcag
     1201 atgaggagca ggagaagggc agcagcagtg agaaggaggg cagtgaagat gagcactcgg
     1261 gcagcgagag tgaacgggag gaaggtgaca gggacgaggc cagtgacaag agtggcagtg
     1321 gtgaggacga gagcagcgag gatgaggccc gggctgcccg tgacaaagag gagatctttg
     1381 gcagtgatgc tgattctgag gacgatgccg actctgatga tgaggacaga ggacaggccc
     1441 aaggtggcag tgacaatgat tcagacagcg gcagcaatgg gggtggccag cggagccgga
     1501 gccacagccg cagcgccagt cccttcccca gtggcagcga gcactcggcc caggaggatg
     1561 gcagtgaagc tgcagcttct gattccagtg aagctgatag tgacagtgac tgaaagcttg
     1621 tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1681 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1741 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1861 cttatcatgt ctggatctga tcactgcttg agcctagaag atccggctgc taacaaagcc
     1921 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactatcata acccctagga
     1981 gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     2041 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     2101 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     2161 tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca
     2221 tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta
     2281 atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga atgggacgcg
     2341 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2401 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2461 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2521 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2581 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct
     2641 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2701 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2761 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     2821 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2881 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2941 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3001 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3061 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3121 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3181 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3241 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3301 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3361 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3421 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3481 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3541 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3601 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3661 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3721 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3781 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3841 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3901 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3961 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4021 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4081 gcagatacca aatactattc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4141 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4201 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4261 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4321 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4381 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4441 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4501 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4561 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4621 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4681 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     4741 tctccttacg catctgtgcg gtatttcaca ccgcatagac cagccgcgta acctggcaaa
     4801 atcggttacg gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg cggacaataa
     4861 agtcttaaac tgaacaaaat agatctaaac tatgacaata aagtcttaaa ctagacagaa
     4921 tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga cttttgttat
     4981 ggctaaagca aactcttcat tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg
     5041 tggccaaggg catggtaaag actatattcg cggcgttgtg acaatttacc gaacaactcc
     5101 gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata
     5161 tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg
     5221 tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg
     5281 aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga
     5341 tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga
     5401 gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc
     5461 aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac
     5521 tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta
     5581 catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc
     5641 tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc
     5701 cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc
     5761 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
     5821 ggaccagttg cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta
     5881 tgtcaactgg gtttgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg
     5941 cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc
     6001 cacgcatcgt caggcattgg cggccttgct gttcttctac ggcaaggtgc tgtgcacgga
     6061 tctgcccttg cttcaggaga tcggtagacc tcggccgtcg cggcgcttgc cggtggtgct
     6121 gaccccggat gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc
     6181 ccaggactct agctatagtt ctagtggttg gctacagctt tgtttgtact atcaacaggt
     6241 tgaactgctg atcaacagat cctctacgcg gccgcggtac cataacttcg tatagcatac
     6301 attatacgaa gttatctggt tttactagta tcgattcgcg acctactccg gaatattaat
     6361 agatcatgga gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt
     6421 tcgtaacagt tttgtaataa aaaaacctat aaatattccg gattattcat accgtccca
//
