LOCUS       sequence-461123-        5516 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 5516)
  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 5516)
  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-His(N)-SSU72
FEATURES             Location/Qualifiers
     source          1..5516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..123
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        286..390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(391..459,460..1251)
                     /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(609..628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1422..2010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1911..1930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2164..2181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2315..2539
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2606..3139)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3328..3333,3334..3350,3351..3356))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(3752..3785)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3833..3924
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3849..3866
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3970..3987
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3994..4017
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    4729..4863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4766..4785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4820..4839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4973..5138
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      join(5322..5516,1..260)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5409..5428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
       61 cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct tgttccaaac
      121 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
      181 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
      241 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      301 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      361 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      421 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      481 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      541 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      601 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
      661 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      721 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
      781 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      841 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
      901 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
      961 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1021 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1081 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1141 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1201 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1261 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1321 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1381 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1441 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1501 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1561 aatactattc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1621 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1681 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1741 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1801 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1861 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1921 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1981 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2041 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2101 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2161 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     2221 catctgtgcg gtatttcaca ccgcatagac cagccgcgta acctggcaaa atcggttacg
     2281 gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg cggacaataa agtcttaaac
     2341 tgaacaaaat agatctaaac tatgacaata aagtcttaaa ctagacagaa tagttgtaaa
     2401 ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga cttttgttat ggctaaagca
     2461 aactcttcat tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg
     2521 catggtaaag actatattcg cggcgttgtg acaatttacc gaacaactcc gcggccggga
     2581 agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca
     2641 tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat
     2701 ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga
     2761 tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat
     2821 agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa
     2881 ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga
     2941 ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga
     3001 aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa
     3061 tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat
     3121 cgttgctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     3181 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
     3241 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     3301 cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg
     3361 gtttgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa
     3421 gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt
     3481 caggcattgg cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgcccttg
     3541 cttcaggaga tcggtagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat
     3601 gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct
     3661 agctatagtt ctagtggttg gctacagctt tgtttgtact atcaacaggt tgaactgctg
     3721 atcaacagat cctctacgcg gccgcggtac cataacttcg tatagcatac attatacgaa
     3781 gttatctggt tttactagta tcgattcgcg acctactccg gaatattaat agatcatgga
     3841 gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt
     3901 tttgtaataa aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc
     3961 ggatccatgc accaccacca ccaccacggt gcacttgaag tcctctttca gggacccccg
     4021 tcgtccccgc tgcgggtggc ggtggtgtgc tcgagcaacc agaaccggag catggaggcg
     4081 cacaacatcc tcagcaaacg gggattcagc gtccgatcct ttggaacagg gactcacgtg
     4141 aagcttccag gaccagctcc cgacaagccc aatgtttatg atttcaaaac cacatatgac
     4201 cagatgtaca atgatcttct taggaaagac aaagaactct atacacagaa tgggatttta
     4261 catatgctgg acagaaataa gagaatcaag ccccggccag aaagattcca gaactgcaaa
     4321 gacctgtttg atctgatcct cacttgcgaa gagagagtgt atgaccaggt ggtggaagat
     4381 ctgaattcca gagaacagga gacctgccag cccgtgcacg tggtcaatgt ggacatccag
     4441 gacaaccacg aggaggccac cctgggggcg tttctcatct gtgagctctg ccagtgtatc
     4501 cagcacacgg aagacatgga gaacgagatc gacgagctgc tgcaggagtt cgaggagaag
     4561 agtggccgca cctttctgca caccgtctgc ttctactgaa agcttgtcga gaagtactag
     4621 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4681 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4861 atctgatcac tgcttgagcc tagaagatcc ggctgctaac aaagcccgaa aggaagctga
     4921 gttggctgct gccaccgctg agcaataact atcataaccc ctaggagatc cgaaccagat
     4981 aagtgaaatc tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc
     5041 tacacccagt tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt
     5101 ccacccctcc cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt
     5161 atgtttttaa tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt
     5221 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
     5281 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     5341 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5401 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5461 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacc
//
