LOCUS       sequence-461127-        8459 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 8459)
  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 8459)
  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-Strep(II)(C)-CTR9
FEATURES             Location/Qualifiers
     source          1..8459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      158..612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(245..264)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     454..475
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        638..742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(743..811,812..1603)
                     /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(961..980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1774..2362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2263..2282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2516..2533
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2667..2891
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2958..3491)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3680..3685,3686..3702,3703..3708))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4104..4137)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4185..4276
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4201..4218
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             7844..7867
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             7868..7891
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     polyA_signal    8024..8158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8061..8080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8115..8134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8268..8433
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc tactttttct
       61 ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt aattgactga
      121 atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
      181 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      241 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      301 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      361 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      421 cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa actggaacaa
      481 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
      541 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      601 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactat
     1921 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2401 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2461 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2521 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     2581 cggtatttca caccgcatag accagccgcg taacctggca aaatcggtta cggttgagta
     2641 ataaatggat gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa
     2701 atagatctaa actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc
     2761 agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc
     2821 attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatggtaa
     2881 agactatatt cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc
     2941 tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct
     3001 tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc
     3061 acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg
     3121 gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca
     3181 ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct
     3241 tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg
     3301 gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca
     3361 agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc
     3421 atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg
     3481 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     3541 ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac
     3601 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     3661 catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggtttgtgc
     3721 cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc
     3781 atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt
     3841 ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct tgcttcagga
     3901 gatcggtaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt
     3961 tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact ctagctatag
     4021 ttctagtggt tggctacagc tttgtttgta ctatcaacag gttgaactgc tgatcaacag
     4081 atcctctacg cggccgcggt accataactt cgtatagcat acattatacg aagttatctg
     4141 gttttactag tatcgattcg cgacctactc cggaatatta atagatcatg gagataatta
     4201 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     4261 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatccat
     4321 gtcgcggggc tccatcgaga ttcccctccg ggacactgac gaggtcattg aacttgactt
     4381 cgatcagtta ccggagggag atgaagttat cagtattctg aaacaggaac acacacaact
     4441 gcacatatgg attgctttgg cgctggaata ctacaagcaa ggaaaaacag aagagtttgt
     4501 aaaattgttg gaagcagcac gtatagatgg caatttggac tatagagacc atgaaaaaga
     4561 ccagatgact tgcttggata cattggcagc gtattatgta caacaggctc ggaaagaaaa
     4621 gaataaggac aataaaaagg atcttattac acaggccacc ttgttgtata caatggccga
     4681 taaaattatt atgtatgatc agaaccattt gttgggaaga gcctgcttct gcctacttga
     4741 gggtgacaaa atggatcaag ctgatgcaca gtttcatttt gtactcaatc agtctccaaa
     4801 taatattcca gcccttcttg gtaaagcttg catttccttc aacaagaagg attacagagg
     4861 agctcttgct tactataaga aagcattgcg tactaaccca ggatgtccag cggaagttcg
     4921 tttaggaatg ggtcattgct ttgtgaaact taacaaactg gaaaaagctc gtctggcatt
     4981 cagcagagcc ctggaactca attccaaatg cgtgggagca ttggttggac tggctgttct
     5041 agaactcaac aataaagagg ctgattccat taaaaatggt gtccagcttc tttccagagc
     5101 ctatactatt gatcctagca accctatggt attgaaccat ttggcaaatc actttttctt
     5161 caaaaaggat tatagtaaag tccagcatct ggccctccat gcattccata atacagaagt
     5221 ggaagctatg caagcagaga gctgctatca gctagctaga tcattccatg ttcaggaaga
     5281 ttatgaccaa gcttttcagt actattatca agccacacag tttgcctcat cctcttttgt
     5341 gctcccattt tttggtttgg gacaaatgta tatttatcga ggtgacaaag aaaatgcatc
     5401 tcagtgcttt gagaaggttt tgaaagctta tcctaataat tacgaaacta tgaaaattct
     5461 cggctctctc tatgctgcct cagaagatca agaaaaacga gatattgcca agggccattt
     5521 gaagaaggtc acagaacagt atcccgatga tgttgaagct tggattgaat tggcacaaat
     5581 cttagaacag actgatatac agggtgccct ttcagcctat ggaacagcaa cacgaatcct
     5641 tcaggagaaa gtgcaggccg atgttcctcc agagattctc aataatgtgg gtgccctcca
     5701 ttttagactt ggaaacctag gggaggctaa gaaatatttt ttggcgtcat tggaccgtgc
     5761 aaaagcagaa gcggaacacg atgagcatta ctataacgcc atttccgtta ctacgtcata
     5821 taatctcgcc aggctatatg aggcgatgtg tgaattccat gaagcagaaa aactgtataa
     5881 aaacatctta cgcgaacatc ctaattatgt tgactgctat ttgcgcctag gagccatggc
     5941 tagagataag ggaaactttt atgaggcttc agattggttt aaggaagctc ttcagattaa
     6001 tcaggatcat ccagatgctt ggtctttgat tggcaatctt catttggcaa aacaagaatg
     6061 gggtcctggg cagaagaagt ttgagaggat attaaaacag ccatccacac agagtgatac
     6121 ctattctatg ctagcccttg gcaacgtgtg gctccaaact ttacatcagc ccacccgaga
     6181 tcgagaaaag gaaaagcgtc atcaagatcg tgctctggcc atctacaaac aagtactcag
     6241 aaatgatgca aagaatctgt atgctgccaa tggcatagga gctgttttgg cccacaaagg
     6301 atattttcgt gaagctcgtg atgtatttgc ccaagtaaga gaagcaacag cagatattag
     6361 tgatgtgtgg ctgaacttag cacacatcta tgtggagcaa aagcagtaca tcagcgccgt
     6421 tcagatgtat gaaaactgcc tccgaaagtt ctataagcac caaaacactg aagttgtact
     6481 ctatttggcc cgggccctct tcaagtgtgg caagttacag gaatgcaaac agactttgct
     6541 gaaggctaga catgtggcac ccagtgatac agttcttatg tttaatgtgg ccttggtcct
     6601 gcaaagatta gctacctctg tcctgaaaga tgaaaaaagt aatctgaagg aagtacttaa
     6661 tgctgtgaaa gaactggagc ttgcacatag atacttcagt tatttgagta aagtgggaga
     6721 taaaatgaga tttgatttgg cccttgctgc tacagaagcc aggcagtgtt ctgacttact
     6781 gagccaggcc cagtaccatg tggcccgggc acgcaaacaa gatgaagaag agcgggagct
     6841 gcgggccaag caagagcaag aaaaggagct gttaaggcag aaacttctta aagaacagga
     6901 agagaaacgt ctcagagaaa aggaagagca aaagaaactt ttggaacagc gggcccagta
     6961 tgtggagaag accaaaaata ttcttatgtt tactggtgag actgaagcaa caaaagagaa
     7021 gaaaagaggt ggtggtggtg gacggcgttc taagaaggga ggagagtttg atgaatttgt
     7081 caatgatgac actgatgatg acctacctat atccaaaaag aagaagagaa gaaagggtag
     7141 tggcagtgaa caagaaggtg aagatgagga gggtggtgag agaaagaaga aaaagaggag
     7201 aagacatcca aagggagaag aaggatctga tgatgatgaa acagaaaatg gccccaaacc
     7261 aaaaaaacga cgtccaccaa aagcagagaa gaaaaaggct cccaagccag aacgtctgcc
     7321 tccatcaatg aagggaaaaa taaaatccaa agccataatt tcatcaagtg atgactcttc
     7381 ggatgaggat aaacttaaaa ttgctgatga aggacatccc aggaacagca acagcaacag
     7441 tgactcagac gaggacgaac aacgaaagaa atgtgcctca tcagagagtg attccgatga
     7501 gaaccagaac aagtctggca gcgaggccgg cagtccccgg aggccacgaa gacagcggtc
     7561 agatcaggac tcagacagtg accagccatc cagaaagaga aggccctccg gttctgagca
     7621 gtctgacaat gaatctgtgc agtcagggag aagccactca ggagtttctg agaacgactc
     7681 tcgcccagct tctccaagtg ccgaatcaga tcacgaatcg gagagaggat ctgataatga
     7741 gggttctggc caaggctctg gaaatgaatc ggaaccagag ggatccaaca atgaggcctc
     7801 agatagaggc tcagaacatg ggtcagatga tagtgacggt gcacttgaag tcctctttca
     7861 gggaccctgg agccacccgc agttcgaaaa gtgaaagctt gtcgagaagt actagaggat
     7921 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     7981 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     8041 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     8101 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg
     8161 atcactgctt gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     8221 ctgctgccac cgctgagcaa taactatcat aacccctagg agatccgaac cagataagtg
     8281 aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac
     8341 ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc
     8401 cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgt
//
