LOCUS       sequence-461133-        6536 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 6536)
  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 6536)
  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)-CDC73
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      345..799
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(432..451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     641..662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        825..929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(930..998,999..1790)
                     /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(1148..1167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1961..2549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2450..2469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2703..2720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2854..3078
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3145..3678)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3867..3872,3873..3889,3890..3895))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4291..4324)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4372..4463
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4388..4405
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4509..4532
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4539..4562
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    6288..6422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6325..6344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6379..6398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  join(6532..6536,1..161)
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat tagcttacga
       61 cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt aaaaactcca
      121 tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat ttttcttcct
      181 gttatgtttt taatcaaaca tcctgccaac tccatgtgac aaaccgtcat cttcggctac
      241 tttttctctg tcacagaatg aaaatttttc tgtcatctct tcgttattaa tgtttgtaat
      301 tgactgaata tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
      361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
      421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
      481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
      541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
      601 ggtttttcgc cctttgacgt tggagtccac gttcttaata gtggactctt gttccaaact
      661 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      721 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      781 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      841 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      901 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      961 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1021 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1081 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1141 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1201 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1261 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1321 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1381 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1441 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1501 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1561 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1621 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1681 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1741 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1801 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1861 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1921 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1981 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2041 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2101 atactattct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2161 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2221 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2281 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2341 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2401 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2461 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2521 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2581 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2641 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2701 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     2761 atctgtgcgg tatttcacac cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg
     2821 ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact
     2881 gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat agttgtaaac
     2941 tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg gctaaagcaa
     3001 actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt ggccaagggc
     3061 atggtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa
     3121 gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat
     3181 cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc
     3241 tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat
     3301 gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata
     3361 gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac
     3421 cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag
     3481 gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa
     3541 aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat
     3601 gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc
     3661 gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
     3721 gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca
     3781 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
     3841 gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg
     3901 tttgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag
     3961 tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc
     4021 aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgcccttgc
     4081 ttcaggagat cggtagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg
     4141 aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta
     4201 gctatagttc tagtggttgg ctacagcttt gtttgtacta tcaacaggtt gaactgctga
     4261 tcaacagatc ctctacgcgg ccgcggtacc ataacttcgt atagcataca ttatacgaag
     4321 ttatctggtt ttactagtat cgattcgcga cctactccgg aatattaata gatcatggag
     4381 ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt
     4441 ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc atcgggcgcg
     4501 gatccatgga ctacaaggac gacgatgaca agctcgatct tgaagtcctc tttcagggac
     4561 ccatggcgga cgtgcttagc gtcctgcgac agtacaacat ccagaagaag gagattgtgg
     4621 tgaagggaga cgaagtgatc ttcggggagt tctcctggcc caagaatgtg aagaccaact
     4681 atgttgtttg ggggactgga aaggaaggcc aacccagaga gtactacaca ttggattcca
     4741 ttttatttct acttaataac gtgcaccttt ctcatcctgt ttatgtccga cgtgcagcta
     4801 ctgaaaatat tcctgtggtt agaagacctg atcgaaaaga tctacttgga tatctcaatg
     4861 gtgaagcgtc aacatcggca agtatagaca gaagcgctcc cttagaaata ggtcttcagc
     4921 gatctactca agtcaaacga gctgcagatg aagttttagc agaagcaaag aaaccacgaa
     4981 ttgaggatga agagtgtgtg cgccttgata aagagagatt ggctgcccgt ttggagggtc
     5041 acaaagaagg gattgtacag actgaacaga ttaggtcttt gtctgaagct atgtcagtgg
     5101 aaaaaattgc tgcaatcaaa gccaaaatta tggctaagaa aagatctact atcaagactg
     5161 atctagatga tgacataact gcccttaaac agaggagttt tgtggatgct gaggtagatg
     5221 tgacccgaga tattgtcagc agagagagag tatggaggac acgaacaact atcttacaaa
     5281 gcacaggaaa gaatttttcc aagaacattt ttgcaattct tcaatctgta aaagccagag
     5341 aagaagggcg tgcacctgaa cagcgacctg ccccaaatgc agcacctgtg gatcccactt
     5401 tgcgcaccaa acagcctatc ccagctgcct ataacagata cgatcaggaa agattcaaag
     5461 gaaaagaaga aacggaaggc ttcaaaattg acactatggg aacctaccat ggtatgacac
     5521 tgaaatctgt aacggagggt gcatctgccc ggaagactca gactcctgca gcccagccag
     5581 taccaagacc agtttctcaa gcaagacctc ccccaaatca gaagaaagga tctcgaacac
     5641 ccattatcat aattcctgca gctaccacct ctttaataac catgcttaat gcaaaagacc
     5701 ttctacagga cctgaaattt gtcccatcag atgaaaagaa gaaacaaggt tgtcaacgag
     5761 aaaatgaaac tctaatacaa agaagaaaag accagatgca accagggggc actgcaatta
     5821 gtgttacagt accttataga gtagtagacc agccccttaa acttatgcct caagactggg
     5881 accgcgttgt agccgttttt gtgcagggtc ctgcatggca gttcaaaggt tggccatggc
     5941 ttttgcctga tggatcacca gttgatatat ttgctaaaat taaagccttc catctgaagt
     6001 atgatgaagt tcgtctggat ccaaatgttc agaaatggga tgtaacagta ttagaactca
     6061 gctatcacaa acgtcatttg gatagaccag tgttcttacg gttttgggaa acattggaca
     6121 ggtacatggt aaagcataaa tcgcacttga gattctgaaa gcttgtcgag aagtactaga
     6181 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     6241 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     6301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     6421 tctgatcact gcttgagcct agaagatccg gctgctaaca aagcccgaaa ggaagctgag
     6481 ttggctgctg ccaccgctga gcaataacta tcataacccc taggagatcc gaacca
//
