LOCUS       sequence-463313-        6289 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 6289)
  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 6289)
  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)-CDK12-KiD(700-1082)
FEATURES             Location/Qualifiers
     source          1..6289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1213..1347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1250..1269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1304..1323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1457..1622
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1806..2260
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1893..1912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2102..2123
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2286..2390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2391..2459,2460..3251)
                     /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(2609..2628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3422..4010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3911..3930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4164..4181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4315..4539
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4606..5139)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5328..5333,5334..5350,5351..5356))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(5752..5785)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5833..5924
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5849..5866
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5970..5993
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6000..6023
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 gaaggaaatt gtcacagata aacaagatgc actggatttc aagaaggaca aaggtgcctt
       61 ttaccttgta tttgagtata tggaccatga cttaatggga ctgctagaat ctggtttggt
      121 gcacttttct gaggaccata tcaagtcgtt catgaaacag ctaatggaag gattggaata
      181 ctgtcacaaa aagaatttcc tgcatcggga tattaagtgt tctaacattt tgctgaataa
      241 cagtgggcaa atcaaactag cagattttgg acttgctcgg ctctataact ctgaagagag
      301 tcgcccttac acaaacaaag tcattacttt gtggtaccga cctccagaac tactgctagg
      361 agaggaacgt tacacaccag ccatagatgt ttggagctgt ggatgtattc ttggggaact
      421 attcacaaag aagcctattt ttcaagccaa tctggaactg gctcagctag aactgatcag
      481 ccgactttgt ggtagccctt gtccagctgt gtggcctgat gttatcaaac tgccctactt
      541 caacaccatg aaaccgaaga agcaatatcg aaggcgtcta cgagaagaat tctctttcat
      601 tccttctgca gcacttgatt tattggacca catgctgaca ctagatccta gtaagcggtg
      661 cacagctgaa cagaccctac agagcgactt ccttaaagat gtcgaactca gcaaaatggc
      721 tcctccagac ctcccccact ggcaggattg ccatgagttg tggagtaaga aacggcgacg
      781 tcagcgacaa agtggtgttg tagtcgaaga gccacctcca tccaaaactt ctcgaaaaga
      841 aactacctca gggacaagta ctgagcctgt gaagaacagc taaaagcttg tcgagaagta
      901 ctgagcctgt gaagaacagc taaaagcttg tcgagaagta ctgagcctgt gaagaacagc
      961 taaaagcttg tcgagaagta ctgagcctgt gaagaacagc taaaagcttg tcgagaagta
     1021 ctgagcctgt gaagaacagc taaaagcttg tcgagaagta ctgagcctgt gaagaacagc
     1081 taaaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt
     1141 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     1201 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1321 atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctagaag atccggctgc
     1381 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactatcata
     1441 acccctagga gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt
     1501 aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct
     1561 aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca
     1621 cagcggggca tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga
     1681 caaaccgtca tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc
     1741 ttcgttatta atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga
     1801 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1861 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1921 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1981 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2041 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttcttaat
     2101 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2161 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2221 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2281 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2341 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2401 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2461 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2521 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2581 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2641 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2701 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2761 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2821 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2881 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2941 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3001 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3061 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3121 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3181 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3241 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3301 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3361 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3421 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3481 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3541 tcagcagagc gcagatacca aatactattc ttctagtgta gccgtagtta ggccaccact
     3601 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3661 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3721 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3781 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3841 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3901 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3961 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4021 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4081 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4141 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4201 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatagac cagccgcgta
     4261 acctggcaaa atcggttacg gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg
     4321 cggacaataa agtcttaaac tgaacaaaat agatctaaac tatgacaata aagtcttaaa
     4381 ctagacagaa tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga
     4441 cttttgttat ggctaaagca aactcttcat tttctgaagt gcaaattgcc cgtcgtatta
     4501 aagaggggcg tggccaaggg catggtaaag actatattcg cggcgttgtg acaatttacc
     4561 gaacaactcc gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact
     4621 tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac
     4681 tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc
     4741 ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga
     4801 gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta
     4861 agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt
     4921 actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac
     4981 gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg
     5041 gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg
     5101 actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc cataacatca
     5161 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtacaa
     5221 aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg
     5281 tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt ttacgaaccg
     5341 aacaggctta tgtcaactgg gtttgtgcct tcatccgttt ccacggtgtg cgtcacccgg
     5401 caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg
     5461 tttcggtctc cacgcatcgt caggcattgg cggccttgct gttcttctac ggcaaggtgc
     5521 tgtgcacgga tctgcccttg cttcaggaga tcggtagacc tcggccgtcg cggcgcttgc
     5581 cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc
     5641 gtttgttcgc ccaggactct agctatagtt ctagtggttg gctacagctt tgtttgtact
     5701 atcaacaggt tgaactgctg atcaacagat cctctacgcg gccgcggtac cataacttcg
     5761 tatagcatac attatacgaa gttatctggt tttactagta tcgattcgcg acctactccg
     5821 gaatattaat agatcatgga gataattaaa atgataacca tctcgcaaat aaataagtat
     5881 tttactgttt tcgtaacagt tttgtaataa aaaaacctat aaatattccg gattattcat
     5941 accgtcccac catcgggcgc ggatccatgg actacaagga cgacgatgac aagctcgatc
     6001 ttgaagtcct ctttcaggga cccatgccaa aaatttgttg tcctcgttat ggagaaagaa
     6061 gacaaacaga aagcgactgg gggaaacgct gtgtggacaa gtttgacatt attgggatta
     6121 ttggagaagg aacctatggc caagtatata aagccaagga caaagacaca ggagaactag
     6181 tggctctgaa gaaggtgaga ctagacaatg agaaagaggg cttcccaatc acagccattc
     6241 gtgaaatcaa aatccttcgt cagttaatcc accgaagtgt tgttaacat
//
