LOCUS       sequence-461121-        6536 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 6536)
  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 6536)
  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)-CDC73-YY/AA(Y290A, Y293A)
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  6..230
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(364..381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(535..1123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(615..634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1294..2085,2086..2154))
                     /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     1917..1936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2155..2259)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2285..2739)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2422..2443)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2633..2652
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  2923..3088
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    3198..3332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3222..3241)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3276..3295
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5058..5081)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(5088..5111)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(5157..5248)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(5215..5232)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    5296..5329
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(5725..5730,5731..5747,5748..5753)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5942..6475
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gccgcgaata tagtctttac catgcccttg gccacgcccc tctttaatac gacgggcaat
       61 ttgcacttca gaaaatgaag agtttgcttt agccataaca aaagtccagt atgctttttc
      121 acagcataac tggactgatt tcagtttaca actattctgt ctagtttaag actttattgt
      181 catagtttag atctattttg ttcagtttaa gactttattg tccgcccaca cccgcttacg
      241 cagggcatcc atttattact caaccgtaac cgattttgcc aggttacgcg gctggtctat
      301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
      361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      961 cggctacact agaagaatag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     1441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     2281 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2341 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2401 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaag aacgtggact
     2461 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2521 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2581 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2641 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2701 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     2761 aaataagcgt tgatattcag tcaattacaa acattaataa cgaagagatg acagaaaaat
     2821 tttcattctg tgacagagaa aaagtagccg aagatgacgg tttgtcacat ggagttggca
     2881 ggatgtttga ttaaaaacat aacaggaaga aaaatgcccc gctgtgggcg gacaaaatag
     2941 ttgggaactg ggaggggtgg aaatggagtt tttaaggatt atttagggaa gagtgacaaa
     3001 atagatggga actgggtgta gcgtcgtaag ctaatacgaa aattaaaaat gacaaaatag
     3061 tttggaacta gatttcactt atctggttcg gatctcctag gggttatgat agttattgct
     3121 cagcggtggc agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcttcta
     3181 ggctcaagca gtgatcagat ccagacatga taagatacat tgatgagttt ggacaaacca
     3241 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3301 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3361 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3421 gtatggctga ttatgatcct ctagtacttc tcgacaagct ttcagaatct caagtgcgat
     3481 ttatgcttta ccatgtacct gtccaatgtt tcccaaaacc gtaagaacac tggtctatcc
     3541 aaatgacgtt tgtgatagct gagttctaat actgttacat cccatttctg aacatttgga
     3601 tccagacgaa cttcatcata cttcagatgg aaggctttaa ttttagcaaa tatatcaact
     3661 ggtgatccat caggcaaaag ccatggccaa cctttgaact gccatgcagg accctgcaca
     3721 aaaacggcta caacgcggtc ccagtcttga ggcataagtt taaggggctg gtctactact
     3781 ctataaggta ctgtaacact aattgcagtg ccccctggtt gcatctggtc ttttcttctt
     3841 tgtattagag tttcattttc tcgttgacaa ccttgtttct tcttttcatc tgatgggaca
     3901 aatttcaggt cctgtagaag gtcttttgca ttaagcatgg ttattaaaga ggtggtagct
     3961 gcaggaatta tgataatggg tgttcgagat cctttcttct gatttggggg aggtcttgct
     4021 tgagaaactg gtcttggtac tggctgggct gcaggagtct gagtcttccg ggcagatgca
     4081 ccctccgtta cagatttcag tgtcatacca tggtaggttc ccatagtgtc aattttgaag
     4141 ccttccgttt cttcttttcc tttgaatctt tcctgatcgg ctctgttagc ggcagctggg
     4201 ataggctgtt tggtgcgcaa agtgggatcc acaggtgctg catttggggc aggtcgctgt
     4261 tcaggtgcac gcccttcttc tctggctttt acagattgaa gaattgcaaa aatgttcttg
     4321 gaaaaattct ttcctgtgct ttgtaagata gttgttcgtg tcctccatac tctctctctg
     4381 ctgacaatat ctcgggtcac atctacctca gcatccacaa aactcctctg tttaagggca
     4441 gttatgtcat catctagatc agtcttgata gtagatcttt tcttagccat aattttggct
     4501 ttgattgcag caattttttc cactgacata gcttcagaca aagacctaat ctgttcagtc
     4561 tgtacaatcc cttctttgtg accctccaaa cgggcagcca atctctcttt atcaaggcgc
     4621 acacactctt catcctcaat tcgtggtttc tttgcttctg ctaaaacttc atctgcagct
     4681 cgtttgactt gagtagatcg ctgaagacct atttctaagg gagcgcttct gtctatactt
     4741 gccgatgttg acgcttcacc attgagatat ccaagtagat cttttcgatc aggtcttcta
     4801 accacaggaa tattttcagt agctgcacgt cggacataaa caggatgaga aaggtgcacg
     4861 ttattaagta gaaataaaat ggaatccaat gtgtagtact ctctgggttg gccttccttt
     4921 ccagtccccc aaacaacata gttggtcttc acattcttgg gccaggagaa ctccccgaag
     4981 atcacttcgt ctcccttcac cacaatctcc ttcttctgga tgttgtactg tcgcaggacg
     5041 ctaagcacgt ccgccatggg tccctgaaag aggacttcaa gatcgagctt gtcatcgtcg
     5101 tccttgtagt ccatggatcc gcgcccgatg gtgggacggt atgaataatc cggaatattt
     5161 ataggttttt ttattacaaa actgttacga aaacagtaaa atacttattt atttgcgaga
     5221 tggttatcat tttaattatc tccatgatct attaatattc cggagtaggt cgcgaatcga
     5281 tactagtaaa accagataac ttcgtataat gtatgctata cgaagttatg gtaccgcggc
     5341 cgcgtagagg atctgttgat cagcagttca acctgttgat agtacaaaca aagctgtagc
     5401 caaccactag aactatagct agagtcctgg gcgaacaaac gatgctcgcc ttccagaaaa
     5461 ccgaggatgc gaaccacttc atccggggtc agcaccaccg gcaagcgccg cgacggccga
     5521 ggtctaccga tctcctgaag caagggcaga tccgtgcaca gcaccttgcc gtagaagaac
     5581 agcaaggccg ccaatgcctg acgatgcgtg gagaccgaaa ccttgcgctc gttcgccagc
     5641 caggacagaa atgcctcgac ttcgctgctg cccaaggttg ccgggtgacg cacaccgtgg
     5701 aaacggatga aggcacaaac ccagttgaca taagcctgtt cggttcgtaa actgtaatgc
     5761 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     5821 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc tatgcctcgg
     5881 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     5941 gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg
     6001 tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat
     6061 gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca
     6121 tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc
     6181 tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaagtttga
     6241 gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca
     6301 gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta
     6361 tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt
     6421 gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt
     6481 cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt cacaac
//
