LOCUS       sequence-461163-        5429 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 5429)
  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 5429)
  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)(N)-PIN1
FEATURES             Location/Qualifiers
     source          1..5429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     207..226
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  497..662
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    772..906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(796..815)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     850..869
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1525..1548)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(1555..1578)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(1624..1715)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1682..1699)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    1763..1796
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(2192..2197,2198..2214,2215..2220)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2409..2942
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  3009..3233
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(3367..3384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3538..4126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3618..3637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4297..5088,5089..5157))
                     /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     4920..4939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5158..5262)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5288..5429,1..313))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(join(5425..5429,1..17))
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttggaacaag agtccactat taagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
       61 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
      121 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
      181 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
      241 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
      301 ctacagggcg cgtcccattc gccattcagg ctgcaaataa gcgttgatat tcagtcaatt
      361 acaaacatta ataacgaaga gatgacagaa aaattttcat tctgtgacag agaaaaagta
      421 gccgaagatg acggtttgtc acatggagtt ggcaggatgt ttgattaaaa acataacagg
      481 aagaaaaatg ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg
      541 agtttttaag gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg
      601 taagctaata cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg
      661 ttcggatctc ctaggggtta tgatagttat tgctcagcgg tggcagcagc caactcagct
      721 tcctttcggg ctttgttagc agccggatct tctaggctca agcagtgatc agatccagac
      781 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
      841 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
      901 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
      961 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagta
     1021 cttctcgaca agctttcact cagtgcggag gatgatgtgg atgccggaat ccgtgaacac
     1081 gggcccgctc atctcccccg tccgcagcgc aaacgaggcg tcttcaaatg gcttctgcat
     1141 ctgacctctg ctgaaggcac ccaggtctcc cctggccttg gctgagctgc agtcgctgaa
     1201 ctgtgaggcc agagactcaa agtcctcctc tcccgacttg atcttctgga tgtagccgtt
     1261 gatcagctcc agggcctcct ccttggtccg ggtgatcttc tcctgccgcc aggacgaggg
     1321 ccgccgtgac tggctgtgct tcaccagcag gtgcgagcag cggaccctgg caggctcccc
     1381 ctgcccgttt ttgccaccac tgctgctgtt gccgctgggc cgctcccact ggctggcgtt
     1441 agtgatgtgg ttgaagtagt acactcggcc tgagctgcgg ctcatgcgct tctcccagcc
     1501 gggcggcagc ttctcctcgt ccgcgggtcc ctgaaagagg acttcaagtg cacccttttc
     1561 gaactgcggg tggctccaca tggatccgcg cccgatggtg ggacggtatg aataatccgg
     1621 aatatttata ggttttttta ttacaaaact gttacgaaaa cagtaaaata cttatttatt
     1681 tgcgagatgg ttatcatttt aattatctcc atgatctatt aatattccgg agtaggtcgc
     1741 gaatcgatac tagtaaaacc agataacttc gtataatgta tgctatacga agttatggta
     1801 ccgcggccgc gtagaggatc tgttgatcag cagttcaacc tgttgatagt acaaacaaag
     1861 ctgtagccaa ccactagaac tatagctaga gtcctgggcg aacaaacgat gctcgccttc
     1921 cagaaaaccg aggatgcgaa ccacttcatc cggggtcagc accaccggca agcgccgcga
     1981 cggccgaggt ctaccgatct cctgaagcaa gggcagatcc gtgcacagca ccttgccgta
     2041 gaagaacagc aaggccgcca atgcctgacg atgcgtggag accgaaacct tgcgctcgtt
     2101 cgccagccag gacagaaatg cctcgacttc gctgctgccc aaggttgccg ggtgacgcac
     2161 accgtggaaa cggatgaagg cacaaaccca gttgacataa gcctgttcgg ttcgtaaact
     2221 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2281 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg tacagtctat
     2341 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     2401 cagcaacgat gttacgcagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     2461 agttaggtgg ctcaagtatg ggcatcattc gcacatgtag gctcggccct gaccaagtca
     2521 aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt cggagacgta gccacctact
     2581 cccaacatca gccggactcc gattacctcg ggaacttgct ccgtagtaag acattcatcg
     2641 cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct cgcggcttac gttctgccca
     2701 agtttgagca gccgcgtagt gagatctata tctatgatct cgcagtctcc ggcgagcacc
     2761 ggaggcaggg cattgccacc gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg
     2821 gtgcttatgt gatctacgtg caagcagatt acggtgacga tcccgcagtg gctctctata
     2881 caaagttggg catacgggaa gaagtgatgc actttgatat cgacccaagt accgccacct
     2941 aacaattcgt tcaagccgag atcggcttcc cggccgcgga gttgttcggt aaattgtcac
     3001 aacgccgcga atatagtctt taccatgccc ttggccacgc ccctctttaa tacgacgggc
     3061 aatttgcact tcagaaaatg aagagtttgc tttagccata acaaaagtcc agtatgcttt
     3121 ttcacagcat aactggactg atttcagttt acaactattc tgtctagttt aagactttat
     3181 tgtcatagtt tagatctatt ttgttcagtt taagacttta ttgtccgccc acacccgctt
     3241 acgcagggca tccatttatt actcaaccgt aaccgatttt gccaggttac gcggctggtc
     3301 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     3361 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3421 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3481 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3541 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3601 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3661 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3721 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3781 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3841 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3901 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3961 ctacggctac actagaagaa tagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4021 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4081 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4141 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4201 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4261 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4321 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4381 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4441 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4501 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4561 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4621 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4681 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4741 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4801 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4861 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4921 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4981 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5041 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5101 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5161 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5221 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5281 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5341 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5401 accgagatag ggttgagtgt tgttccagt
//
