LOCUS       sequence-461158-        6626 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 6626)
  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 6626)
  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-Cyclin K
FEATURES             Location/Qualifiers
     source          1..6626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     297..318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        481..585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(586..654,655..1446)
                     /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(804..823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1617..2205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2106..2125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2359..2376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2510..2734
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2801..3334)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3523..3528,3529..3545,3546..3551))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(3947..3980)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4028..4119
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4044..4061
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    6034..6168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6071..6090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6125..6144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6278..6443
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc ttaatagtgg
      301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
      361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
      421 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
      481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1741 agagcgcaga taccaaatac tattcttcta gtgtagccgt agttaggcca ccacttcaag
     1801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2281 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2341 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2401 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagaccagcc gcgtaacctg
     2461 gcaaaatcgg ttacggttga gtaataaatg gatgccctgc gtaagcgggt gtgggcggac
     2521 aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc ttaaactaga
     2581 cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata ctggactttt
     2641 gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag
     2701 gggcgtggcc aagggcatgg taaagactat attcgcggcg ttgtgacaat ttaccgaaca
     2761 actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg gtacttgggt
     2821 cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga gccactgcgg
     2881 gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg ttggcctcat
     2941 gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg ccggagactg
     3001 cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggcaga acgtaagccg
     3061 cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat gtcttactac
     3121 ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg gctacgtctc
     3181 cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg tcagggccga
     3241 gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta gggcgactgc
     3301 cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc tgctccataa catcaaacat
     3361 cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa
     3421 cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag
     3481 gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagtttacg aaccgaacag
     3541 gcttatgtca actgggtttg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc
     3601 ttgggcagca gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg caaggtttcg
     3661 gtctccacgc atcgtcaggc attggcggcc ttgctgttct tctacggcaa ggtgctgtgc
     3721 acggatctgc ccttgcttca ggagatcggt agacctcggc cgtcgcggcg cttgccggtg
     3781 gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg
     3841 ttcgcccagg actctagcta tagttctagt ggttggctac agctttgttt gtactatcaa
     3901 caggttgaac tgctgatcaa cagatcctct acgcggccgc ggtaccataa cttcgtatag
     3961 catacattat acgaagttat ctggttttac tagtatcgat tcgcgaccta ctccggaata
     4021 ttaatagatc atggagataa ttaaaatgat aaccatctcg caaataaata agtattttac
     4081 tgttttcgta acagttttgt aataaaaaaa cctataaata ttccggatta ttcataccgt
     4141 cccaccatcg ggcgcggatc catgaaggag aataaagaaa attcaagccc ttcagtaact
     4201 tcagcaaacc tggaccacac aaagccatgt tggtactggg ataagaaaga cttggctcat
     4261 acaccctcac aacttgaagg acttgatcca gccaccgagg cccggtaccg ccgagagggc
     4321 gctcggttca tctttgatgt gggcacacgt ttggggctac actatgatac cctggcaact
     4381 ggaataattt attttcatcg cttctatatg tttcattcct tcaagcaatt cccaagatat
     4441 gtgacaggag cctgttgcct ctttctggct gggaaagtag aagaaacacc aaaaaaatgt
     4501 aaagatatca tcaaaacagc tcgtagttta ttaaatgatg tacaatttgg ccagtttgga
     4561 gatgacccaa aggaggaagt aatggttctg gagagaatct tactgcagac catcaagttt
     4621 gatttacagg tagaacatcc ataccagttc ctactaaaat atgcaaagca actcaaaggt
     4681 gataaaaaca aaattcaaaa gttggttcaa atggcatgga catttgtaaa tgacagtctc
     4741 tgcaccacct tgtcactgca gtgggaacca gagatcatag cagtagcagt gatgtatctc
     4801 gcaggacgtt tgtgcaaatt tgaaatacaa gaatggacct ccaaacccat gtataggaga
     4861 tggtgggagc agtttgttca agatgtcccg gtcgacgttt tggaagacat ctgccaccaa
     4921 atcctggatc tttactcaca aggaaaacaa cagatgcctc atcacacccc ccatcagctg
     4981 caacagcccc catctcttca gcctacacca caagtgccgc aagtacagca gtcacagccg
     5041 tctcaaagct ccgaaccatc ccagccccag cagaaggacc cccagcaacc agcccagcag
     5101 cagcagccag cccaacagcc caagaaaccc tctccgcagc ccagttctcc ccgacaggtt
     5161 aagcgagccg tggttgtttc tcccaaagaa gagaacaaag cagcagaacc accaccacct
     5221 aaaatcccca aaattgagac cactcatcca ccgttgcctc cagcccaccc acctccagac
     5281 cggaagcctc ccctcgctgc tgccttaggt gaggctgagc cgccgggccc tgtggatgcc
     5341 actgacctcc ccaaagtcca gattccccct ccggcccacc cggcccctgt gcaccagcca
     5401 ccgccgctgc cacaccggcc cccgccccca cccccctcca gctacatgac cgggatgtcc
     5461 accaccagct cctacatgtc tggagagggc taccagagcc tgcagtccat gatgaagacc
     5521 gagggaccct cctacggtgc cctgcccccc gcctacggcc cacctgcaca cctgccctac
     5581 cacccccatg tctacccgcc caacccgccc ccgccacctg tgcctcctcc cccagcctcc
     5641 ttccccccac ctgccatccc accccctact cctggctacc ccccaccccc acccacctac
     5701 aaccccaact tcccaccccc acccccacgc ctcccgccta cccacgcagt cccccctcat
     5761 cctcctccag ggttgggcct gccgccagcc agctacccac ctcctgccgt cccccctgga
     5821 ggacagcctc ctgtgccccc gcccattccc ccacccggca tgcctccagt tggggggctg
     5881 gggcgggcag cctggatgag ataaaagctt gtcgagaagt actagaggat cataatcagc
     5941 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     6001 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     6061 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6121 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt
     6181 gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     6241 cgctgagcaa taactatcat aacccctagg agatccgaac cagataagtg aaatctagtt
     6301 ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca
     6361 tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt
     6421 cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa
     6481 catcctgcca actccatgtg acaaaccgtc atcttcggct actttttctc tgtcacagaa
     6541 tgaaaatttt tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct
     6601 tatttgcagc ctgaatggcg aatggg
//
