LOCUS       sequence-463629-        8582 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 8582)
  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 8582)
  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)-CDK12deltaRS(delta104-380)
FEATURES             Location/Qualifiers
     source          1..8582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      287..741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(374..393)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        767..871
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(872..940,941..1732)
                     /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(1090..1109)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1903..2491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2392..2411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2645..2662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2796..3020
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3087..3620)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3809..3814,3815..3831,3832..3837))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4233..4266)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4314..4405
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4330..4347
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4451..4474
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4481..4504
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    8276..8410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8313..8332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8367..8386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  join(8520..8582,1..103)
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgctacac ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc
       61 catttccacc cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc
      121 ctgttatgtt tttaatcaaa catcctgcca actccatgtg acaaaccgtc atcttcggct
      181 actttttctc tgtcacagaa tgaaaatttt tctgtcatct cttcgttatt aatgtttgta
      241 attgactgaa tatcaacgct tatttgcagc ctgaatggcg aatgggacgc gccctgtagc
      301 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      361 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      421 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      481 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
      541 acggtttttc gccctttgac gttggagtcc acgttcttaa tagtggactc ttgttccaaa
      601 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
      661 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      721 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
      781 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      841 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
      901 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
      961 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1021 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1081 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1141 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1201 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1261 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1321 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1381 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1441 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1501 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1561 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     1621 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     1681 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     1741 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     1801 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1861 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2041 aaatactatt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2161 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2221 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2281 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2341 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2401 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2461 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2521 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2581 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2641 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2701 gcatctgtgc ggtatttcac accgcataga ccagccgcgt aacctggcaa aatcggttac
     2761 ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa
     2821 ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa
     2881 actgaaatca gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc
     2941 aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg
     3001 gcatggtaaa gactatattc gcggcgttgt gacaatttac cgaacaactc cgcggccggg
     3061 aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat atcaaagtgc
     3121 atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc gtcaccgtaa
     3181 tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt gaggagattg
     3241 atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag atcatagata
     3301 tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca
     3361 accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag caagttcccg
     3421 aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa ctcacgaccg
     3481 aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct acatgtgcga
     3541 atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg ctgcgtaaca
     3601 tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     3661 acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc
     3721 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     3781 gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg
     3841 ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga
     3901 agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg
     3961 tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctt
     4021 gcttcaggag atcggtagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga
     4081 tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc
     4141 tagctatagt tctagtggtt ggctacagct ttgtttgtac tatcaacagg ttgaactgct
     4201 gatcaacaga tcctctacgc ggccgcggta ccataacttc gtatagcata cattatacga
     4261 agttatctgg ttttactagt atcgattcgc gacctactcc ggaatattaa tagatcatgg
     4321 agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt ttcgtaacag
     4381 ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca ccatcgggcg
     4441 cggatccatg gactacaagg acgacgatga caagctcgat cttgaagtcc tctttcaggg
     4501 acccatgccc aattcagaga gacatggggg caagaaggac gggagtggag gagcttctgg
     4561 aactttgcag ccgtcatcgg gaggcggcag ctctaacagc agagagcgtc accgcttggt
     4621 atcgaagcac aagcggcata agtccaaaca ctccaaagac atggggttgg tgacccccga
     4681 agcagcatcc ctgggcacag ttatcaaacc tttggtggag tatgatgata tcagctctga
     4741 ttccgacacc ttctccgatg acatggcctt caaactagac cgaagggaga acgacgaacg
     4801 tcgtggatca gatcattcca gtatctcacc tgtcaggctt ccacttaatt ccagtctggg
     4861 agctgaactc agtaggaaaa agaaggaaag agcagctgct gctgctgcag caaagatgga
     4921 tggaaaggag tccaagggtt cacctgtatt tttgcctaga aaagagaaca gttcagtaga
     4981 ggctaaggat tcaggtttgg agtctaaaaa gttacccaga agtgtaaaat tggaaaaatc
     5041 tgccccagat actgaactgg tgaatgtaac acatctaaac acagaggtaa aaaattcttc
     5101 agatacaggg aaagtaaagt tggatgagaa ctccgagaag catcttgtta aagatttgaa
     5161 agcacaggga acaagagact ctaaacccat agcactgaaa gaggagattg ttactccaaa
     5221 ggagacagaa acatcagaaa aggagacccc tccacctctt cccacaattg cttctccccc
     5281 accccctcta ccaactacta cccctccacc tcagacaccc cctttgccac ctttgcctcc
     5341 aataccagct cttccacagc aaccacctct gcctccttct cagccagcat ttagtcaggt
     5401 tcctgcttcc agtacttcaa ctttgccccc ttctactcac tcaaagacat ctgctgtgtc
     5461 ctctcaggca aattctcagc cccctgtaca ggtttctgtg aagactcaag tatctgtaac
     5521 agctgctatt ccacacctga aaacttcaac gttgcctcct ttgcccctcc cacccttatt
     5581 acctggagat gatgacatgg atagtccaaa agaaactctt ccttcaaaac ctgtgaagaa
     5641 agagaaggaa cagaggacac gtcacttact cacagacctt cctctccctc cagagctccc
     5701 tggtggagat ctgtctcccc cagactctcc agaaccaaag gcaatcacac cacctcagca
     5761 accatataaa aagagaccaa aaatttgttg tcctcgttat ggagaaagaa gacaaacaga
     5821 aagcgactgg gggaaacgct gtgtggacaa gtttgacatt attgggatta ttggagaagg
     5881 aacctatggc caagtatata aagccaagga caaagacaca ggagaactag tggctctgaa
     5941 gaaggtgaga ctagacaatg agaaagaggg cttcccaatc acagccattc gtgaaatcaa
     6001 aatccttcgt cagttaatcc accgaagtgt tgttaacatg aaggaaattg tcacagataa
     6061 acaagatgca ctggatttca agaaggacaa aggtgccttt taccttgtat ttgagtatat
     6121 ggaccatgac ttaatgggac tgctagaatc tggtttggtg cacttttctg aggaccatat
     6181 caagtcgttc atgaaacagc taatggaagg attggaatac tgtcacaaaa agaatttcct
     6241 gcatcgggat attaagtgtt ctaacatttt gctgaataac agtgggcaaa tcaaactagc
     6301 agattttgga cttgctcggc tctataactc tgaagagagt cgcccttaca caaacaaagt
     6361 cattactttg tggtaccgac ctccagaact actgctagga gaggaacgtt acacaccagc
     6421 catagatgtt tggagctgtg gatgtattct tggggaacta ttcacaaaga agcctatttt
     6481 tcaagccaat ctggaactgg ctcagctaga actgatcagc cgactttgtg gtagcccttg
     6541 tccagctgtg tggcctgatg ttatcaaact gccctacttc aacaccatga aaccgaagaa
     6601 gcaatatcga aggcgtctac gagaagaatt ctctttcatt ccttctgcag cacttgattt
     6661 attggaccac atgctgacac tagatcctag taagcggtgc acagctgaac agaccctaca
     6721 gagcgacttc cttaaagatg tcgaactcag caaaatggct cctccagacc tcccccactg
     6781 gcaggattgc catgagttgt ggagtaagaa acggcgacgt cagcgacaaa gtggtgttgt
     6841 agtcgaagag ccacctccat ccaaaacttc tcgaaaagaa actacctcag ggacaagtac
     6901 tgagcctgtg aagaacagca gcccagcacc acctcagcct gctcctggca aggtggagtc
     6961 tggggctggg gatgcaatag gccttgctga catcacacaa cagctgaatc aaagtgaatt
     7021 ggcagtgtta ttaaacctgc tgcagagcca aaccgacctg agcatccctc aaatggcaca
     7081 gctgcttaac atccactcca acccagagat gcagcagcag ctggaagccc tgaaccaatc
     7141 catcagtgcc ctgacggaag ctacttccca gcagcaggac tcagagacca tggccccaga
     7201 ggagtctttg aaggaagcac cctctgcccc agtgatcctg ccttcagcag aacagacgac
     7261 ccttgaagct tcaagcacac cagctgacat gcagaatata ttggcagttc tcttgagtca
     7321 gctgatgaaa acccaagagc cagcaggcag tctggaggaa aacaacagtg acaagaacag
     7381 tgggccacag gggccccgaa gaactcccac aatgccacag gaggaggcag cagcatgtcc
     7441 tcctcacatt cttccaccag agaagaggcc ccctgagccc cccggacctc caccgccgcc
     7501 acctccaccc cctctggttg aaggcgatct ttccagcgcc ccccaggagt tgaacccagc
     7561 cgtgacagcc gccttgctgc aacttttatc ccagcctgaa gcagagcctc ctggccacct
     7621 gccacatgag caccaggcct tgagaccaat ggagtactcc acccgacccc gtccaaacag
     7681 gacttatgga aacactgatg ggcctgaaac agggttcagt gccattgaca ctgatgaacg
     7741 aaactctggt ccagccttga cagaatcctt ggtccagacc ctggtgaaga acaggacctt
     7801 ctcaggctct ctgagccacc ttggggagtc cagcagttac cagggcacag ggtcagtgca
     7861 gtttccaggg gaccaggacc tccgttttgc cagggtcccc ttagcgttac acccggtggt
     7921 cgggcaacca ttcctgaagg ctgagggaag cagcaattct gtggtacatg cagagaccaa
     7981 attgcaaaac tatggggagc tggggccagg aaccactggg gccagcagct caggagcagg
     8041 ccttcactgg gggggcccaa ctcagtcttc tgcttatgga aaactctatc gggggcctac
     8101 aagagtccca ccaagagggg gaagagggag aggagttcct tactaaaagc ttgtcgagaa
     8161 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     8221 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     8281 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     8341 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     8401 tgtctggatc tgatcactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg
     8461 aagctgagtt ggctgctgcc accgctgagc aataactatc ataaccccta ggagatccga
     8521 accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt
     8581 ac
//
