LOCUS       sequence-461135-        6752 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 6752)
  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 6752)
  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-GST(N)-RPB1-CTD(1586-1971)
FEATURES             Location/Qualifiers
     source          1..6752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    510..644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(547..566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     601..620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  754..919
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1103..1557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1190..1209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1399..1420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1583..1687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1688..1756,1757..2548)
                     /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(1906..1925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2719..3307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3208..3227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3461..3478
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3612..3836
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3903..4436)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4625..4630,4631..4647,4648..4653))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(5049..5082)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5130..5221
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5146..5163
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5267..5290
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5297..5950
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     5908..5930
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             5951..5974
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 cttacagtcc cagctcccca gagtataccc caacctctcc caagtactca cctaccagtc
       61 ccaaatattc acccacctct cccaagtact cgcctaccag tcccacctat tcacccacca
      121 ccccaaaata ctccccaaca tctcctactt attccccaac ctctccagtc tacaccccaa
      181 cctctcccaa gtactcacct actagcccca cttactcgcc cacttccccc aagtactcgc
      241 ccaccagccc cacctactcg cccacctccc ccaaaggctc aacctactct cccacttccc
      301 ctggttactc gcccaccagc cccacctaca gtctcacaag cccggctatc agcccggatg
      361 acagtgacga ggagaactga aagcttgtcg agaagtacta gaggatcata atcagccata
      421 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
      481 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
      541 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      601 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
      661 ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
      721 gagcaataac tatcataacc cctaggagat ccgaaccaga taagtgaaat ctagttccaa
      781 actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta
      841 ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca
      901 actattttgt ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc
      961 ctgccaactc catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa
     1021 aatttttctg tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt
     1081 tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1141 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1201 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1261 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1321 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1381 gagtccacgt tcttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1441 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1501 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     1561 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1621 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1681 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1741 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1801 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1861 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1921 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1981 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2041 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2101 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2161 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2221 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2281 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2341 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2401 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2461 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2521 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2581 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2641 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2701 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2761 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2821 tccgaaggta actggcttca gcagagcgca gataccaaat actattcttc tagtgtagcc
     2881 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2941 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3001 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3061 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3121 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3181 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3241 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3301 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3361 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3421 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3481 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3541 catagaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa tggatgccct
     3601 gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga tctaaactat
     3661 gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc agttatgctg
     3721 tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt ctgaagtgca
     3781 aattgcccgt cgtattaaag aggggcgtgg ccaagggcat ggtaaagact atattcgcgg
     3841 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     3901 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     3961 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     4021 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     4081 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     4141 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     4201 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     4261 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     4321 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     4381 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt
     4441 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     4501 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     4561 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     4621 ttacagttta cgaaccgaac aggcttatgt caactgggtt tgtgccttca tccgtttcca
     4681 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc tgtcctggct
     4741 ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg ccttgctgtt
     4801 cttctacggc aaggtgctgt gcacggatct gcccttgctt caggagatcg gtagacctcg
     4861 gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca tcctcggttt
     4921 tctggaaggc gagcatcgtt tgttcgccca ggactctagc tatagttcta gtggttggct
     4981 acagctttgt ttgtactatc aacaggttga actgctgatc aacagatcct ctacgcggcc
     5041 gcggtaccat aacttcgtat agcatacatt atacgaagtt atctggtttt actagtatcg
     5101 attcgcgacc tactccggaa tattaataga tcatggagat aattaaaatg ataaccatct
     5161 cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa aacctataaa
     5221 tattccggat tattcatacc gtcccaccat cgggcgcgga tccatggact acaaggacga
     5281 cgatgacaag ctcgatatgt cccctatact aggttattgg aaaattaagg gccttgtgca
     5341 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
     5401 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
     5461 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
     5521 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
     5581 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
     5641 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
     5701 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
     5761 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
     5821 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
     5881 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
     5941 tcctccaaaa cttgaagtcc tctttcaggg acccggtggt gccatgtctc ccagctactc
     6001 gccaacgtca cctgcctacg agccccgctc tcctgggggc tacacacccc agagtccctc
     6061 ttattccccc acttcaccct cctactcccc tacctctcca tcctattctc caaccagtcc
     6121 caactatagt cccacatcac ccagctattc gccaacgtca cccagctact caccgacctc
     6181 tcccagctac tcacccacct ctcccagcta ctcgcccacc tctcccagct actcgcccac
     6241 ctctcccagc tactcaccca cttcccctag ctactcgccc acttccccta gctactcgcc
     6301 aacgtctccc agctactcgc cgacatctcc cagctactcg ccaacttcac ccagctattc
     6361 tcccacttct cccagctact cacctacctc tccaagctat tcacccacct cccccagcta
     6421 ctcacccact tccccaagtt actcacccac cagcccgaac tattctccaa ccagtcccaa
     6481 ttacacccca acatcaccca gctacagccc gacatcaccc agctattcac ctactagtcc
     6541 caactacaca cctaccagcc ctaactacag cccaacctct ccaagctact ctccaacatc
     6601 acccagctat tccccgacct caccaagtta ctccccttcc agcccacgat acacaccaca
     6661 gtctccaacc tataccccaa gctcacccag ctacagcccc agctcgccca gctacagccc
     6721 aacctcaccc aagtacaccc caaccagtcc tt
//
