LOCUS       sequence-463310-        9296 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 9296)
  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 9296)
  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)-CDK13
FEATURES             Location/Qualifiers
     source          1..9296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2529..2540
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4478..4612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4515..4534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4569..4588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4722..4887
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5071..5525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5158..5177)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5367..5388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5551..5655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5656..5724,5725..6516)
                     /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(5874..5893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6687..7275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7176..7195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7429..7446
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  7580..7804
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7871..8404)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(8593..8598,8599..8615,8616..8621))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(9017..9050)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        9098..9189
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     9114..9131
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             9235..9258
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9265..9288
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 gtcagacacg gccctcggtg gcggtggagg attgtcatgg gccgagaaga aactcgaaga
       61 gagacgcaag cgtaggcgct tcttgtcgcc tcaacagcca cccttgttgc tccctctgtt
      121 gcagcctcaa ctgttgcagc cccctccccc tccaccgcct ctcttgttcc tggctgcacc
      181 aggtacagcg gccgcggccg cagctgctgc cgcagcctct tccagctgct tctcccccgg
      241 tcctccactg gaggtgaaac gtttggccag aggaaaaagg agggcaggag gccgtcaaaa
      301 gcgcagacgt ggtccaagag ctggacaaga ggctgaaaag cgccgtgttt tctcgctgcc
      361 tcagccgcaa caggacggcg gaggtggcgc tagctcaggc ggaggtgtca ccccattggt
      421 cgagtacgaa gacgtgtcga gccaatctga gcaaggcttg ctgttgggtg gcgcttcagc
      481 cgcaaccgca gccacagctg caggcggcac aggaggtagt ggaggctcac ccgcctcaag
      541 ctccggtaca cagcgccgtg gagagggaag cgagcgcagg ccccgccggg accgccgcag
      601 cagcagtggc cgcagcaagg agcgccaccg cgagcaccgg cggcgggatg ggcagcgcgg
      661 tggcagcgag gcctccaagt cccgcagccg ccacagccac agcggcgagg aacgggccga
      721 ggtcgccaag agcggcagca gcagcagcag cggcggccgc cggaaaagcg cttcggccac
      781 atccagcagc agtagcagcc gcaaggaccg ggactcgaag gcccaccgca gccggactaa
      841 gtcgtccaag gagccgcctt cggcctacaa ggaaccgccc aaggcctacc gggaggacaa
      901 gaccgagcct aaggcctaca ggcggcggcg gtccctcagc ccactgggag gccgggacga
      961 cagcccggtg tcccacaggg cctctcagag cctgaggagc cgcaagtccc ccagcccggc
     1021 aggaggtggc agcagcccct attctcggcg gctgccgcgc tccccgagcc cctacagtcg
     1081 ccgccgctcc cccagctaca gccgccacag ctcctacgag cggggcggcg acgtgtcccc
     1141 tagtccctac agcagcagca gctggcgccg ctctcgcagt ccctacagcc ctgtgctcag
     1201 acggtctgga aaatcccgaa gcagaagccc gtattcatct aggcattcaa gatctcgtag
     1261 caggcacaga ttgtctagat ccagaagtcg tcattctagt atttctccta gcacactaac
     1321 tctgaagagt agcctggcag ctgaattgaa caagaataaa aaagcacgag cagcagaggc
     1381 agcgagagcc gcagaagcag cgaaagctgc agaagcaact aaggctgctg aggctgctgc
     1441 caaggctgca aaagcttcaa acacttctac acctaccaag gggaacacgg aaactagtgc
     1501 cagtgcatca caaacaaacc atgtgaagga tgtgaagaaa attaaaattg aacatgcacc
     1561 ttctccctca agtggtggaa ctttaaaaaa tgacaaagca aaaacaaagc cacctcttca
     1621 ggtaacgaag gtggaaaata atttgattgt agataaagcc accaagaaag cagtcatagt
     1681 tggaaaggag agtaaatctg ctgctacaaa ggaggaatca gtatctctta aagagaaaac
     1741 caaaccactt acaccaagca taggagccaa ggagaaggag caacatgtag ctttagtcac
     1801 ctctacatta ccaccgttac ctttgcctcc catgctgcct gaagataaag aagctgatag
     1861 cttacgagga aatatttcag taaaagcagt taaaaaagaa gtagaaaaga aactccgatg
     1921 tcttcttgct gatttaccgc tgccccctga gctaccagga ggagatgatc tttcaaagag
     1981 tccagaggaa aagaaaacag caacacagtt acatagtaaa aggaggccta aaatatgtgg
     2041 gcctcgctat ggtgaaacca aagaaaaaga tattgactgg ggaaaacgct gcgtggataa
     2101 atttgatatc atcggaatta ttggagaagg tacttacgga caagtttaca aagccaggga
     2161 taaagacact ggagaaatgg tagccttaaa aaaagtacgt ctggataatg aaaaggaagg
     2221 ctttccaatt acagcaattc gagaaattaa aattctccgg cagcttaccc atcagagtat
     2281 tatcaatatg aaggaaatag tgactgataa agaagatgct ttggatttca agaaggacaa
     2341 aggtgcattt tatctggtgt ttgaatatat ggaccatgat ctgatgggac tactggaatc
     2401 aggcttggtt cattttaatg aaaatcacat aaagtcattt atgagacagc tcatggaggg
     2461 tctggattat tgtcataaga agaacttttt gcatagagat attaaatgtt ccaatatcct
     2521 tctaaataat agagggcaga taaaacttgc agactttgga cttgctcgat tgtatagctc
     2581 agaagaaagt cggccgtata ctaacaaggt aattacttta tggtaccgtc cacctgaact
     2641 gctactggga gaagaacgat acacaccagc cattgatgta tggagctgtg gctgtatcct
     2701 tggcgaactc ttcactaaaa aacctatatt tcaagcaaat caggaacttg cacaactaga
     2761 attaataagc cgaatatgtg ggagtccatg tcctgcagtg tggcctgatg taatcaaact
     2821 accatatttc aacaccatga aaccaaagaa gcaatatcgt cgaaagttaa gagaagaatt
     2881 tgtttttatt cctgcagctg cgctagactt atttgattac atgcttgcct tggatcctag
     2941 taagcgctgc actgctgaac aggctcttca gtgcgagttc ctccgagatg tggaaccctc
     3001 aaaaatgcct ccaccagatc tccctttatg gcaagattgt catgagttat ggagtaaaaa
     3061 gcgaagaaga cagaagcaga tgggcatgac tgatgatgtt tccacaatta aagcccccag
     3121 gaaggacttg tctctgggct tggatgacag cagaaccaac acaccccagg gtgtgctgcc
     3181 atcttcacag ctgaaatctc agggcagctc aaatgtggca cctggtgaaa aacagacaga
     3241 tccatcaaca ccacaacagg agtcttcgaa accgttggga ggaattcagc cttcttctca
     3301 gaccatccag cctaaagtgg agactgatgc tgcccaggcg gctgtgcaga gtgcatttgc
     3361 agttctgttg actcagttaa taaaggctca gcagtcaaag cagaaagatg tgctactaga
     3421 agagagggaa aatggatcgg gacatgaagc gtcattacaa ctcaggccac ctccagaacc
     3481 tagcactccg gtgtcgggac aagatgacct catccagcat caagatatga ggatcttgga
     3541 gctaacgcca gaaccagacc ggcctcgaat tctgcctcct gaccaacgac ctcccgagcc
     3601 tcctgaacca ccaccagtca ctgaggaaga tctagattat cggacagaaa accagcatgt
     3661 acccaccacc agttcttcat taactgaccc tcatgccgga gtgaaggcag ccctgttaca
     3721 gctgcttgct cagcatcagc cccaggatga ccccaaaaga gaaggtggga ttgattatca
     3781 agcaggagac acttacgtgt ccacttcaga ctacaaggac aactttggat cctcttcttt
     3841 ctcttctgct ccttatgtta gcaatgatgg tctaggaagc agttctgctc caccactaga
     3901 acgacgtagt ttcattggaa attcagatat tcagtctttg gataactaca gtactgcttc
     3961 atctcattct ggtggtccac ctcagccttc tgccttttct gagtcatttc ccagttcagt
     4021 agctggatat ggagacattt acctcaatgc tggtcccatg ttgtttagtg gagacaagga
     4081 ccatagattt gaatatagcc atggtcctat tgcagtcctg gcaaacagca gtgacccttc
     4141 cacggggcca gagagtactc atcctttgcc agcaaagatg cacaactata actatggtgg
     4201 taacttacag gaaaatccga gtggccccag cctcatgcat ggacagacct ggacttctcc
     4261 tgcccaagga cctggatatt cacaaggata caggggacat attagcacat caactggcag
     4321 aggcagaggc agagggttac catactgaaa gcttgtcgag aagtactaga ggatcataat
     4381 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4441 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     4621 gcttgagcct agaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     4681 ccaccgctga gcaataacta tcataacccc taggagatcc gaaccagata agtgaaatct
     4741 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     4801 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     4861 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat
     4921 caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac
     4981 agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa
     5041 cgcttatttg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     5101 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5161 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5221 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5281 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5341 tgacgttgga gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa
     5401 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5461 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5521 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5581 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5641 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5701 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5761 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5821 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5881 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5941 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6001 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6061 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6121 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6181 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6241 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6301 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6361 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6421 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6481 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6541 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6601 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6661 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6721 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6781 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tattcttcta
     6841 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6901 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6961 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7021 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7081 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7141 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7201 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7261 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7321 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7381 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7441 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     7501 tcacaccgca tagaccagcc gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg
     7561 gatgccctgc gtaagcgggt gtgggcggac aataaagtct taaactgaac aaaatagatc
     7621 taaactatga caataaagtc ttaaactaga cagaatagtt gtaaactgaa atcagtccag
     7681 ttatgctgtg aaaaagcata ctggactttt gttatggcta aagcaaactc ttcattttct
     7741 gaagtgcaaa ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg taaagactat
     7801 attcgcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt
     7861 gaacgaattg ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta
     7921 tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga
     7981 tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa
     8041 tgccctgcct ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg
     8101 gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga
     8161 aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg
     8221 gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag
     8281 cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg
     8341 agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta
     8401 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
     8461 gatgcccgag gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact
     8521 gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc
     8581 tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggtttg tgccttcatc
     8641 cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg
     8701 tcctggctgg cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc
     8761 ttgctgttct tctacggcaa ggtgctgtgc acggatctgc ccttgcttca ggagatcggt
     8821 agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc
     8881 ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt
     8941 ggttggctac agctttgttt gtactatcaa caggttgaac tgctgatcaa cagatcctct
     9001 acgcggccgc ggtaccataa cttcgtatag catacattat acgaagttat ctggttttac
     9061 tagtatcgat tcgcgaccta ctccggaata ttaatagatc atggagataa ttaaaatgat
     9121 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa
     9181 cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc catggactac
     9241 aaggacgacg atgacaagct cgatcttgaa gtcctctttc agggaccccc aagttc
//
