LOCUS       sequence-469217-        4663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB13 transposase, for use with Sleeping Beauty transponson plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4663)
  AUTHORS   Chan ASL, Zhu H, Narita M, Cassidy LD, Young ARJ, Bermejo-Rodriguez 
            C, Janowska AT, Chen HC, Gough S, Oshimori N, Zender L, Aitken SJ, 
            Hoare M, Narita M
  TITLE     Titration of RAS alters senescent state and influences tumour 
            initiation.
  JOURNAL   Nature. 2024 Sep;633(8030):678-685. doi: 10.1038/s41586-024-07797-z.
            Epub 2024 Aug 7.
  PUBMED    39112713
REFERENCE   2  (bases 1 to 4663)
  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: 
            "10.1038/s41586-024-07797-z"; journalName: "Nature"; date: 
            "2024-09"; volume: "633"; issue: "8030"; pages: "678-685"
COMMENT     Sequence Label: pPGK-SB13
FEATURES             Location/Qualifiers
     source          1..4663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     202..224
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     216..233
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     217..233
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        269..768
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     701..720
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     intron          836..932
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             999..2021
                     /codon_start=1
                     /product="hyperactive variant of the Sleeping Beauty 
                     transposase (Mates et al., 2009)"
                     /label=SB100X
                     /translation="MGKSKEISQDLRKKIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTAAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGRSARKKPLLQNRHKKARLRFARAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMRKENYVDIL
                     KQHLKTSVRKLKLGRKWVFQMDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPTYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     polyA_signal    2035..2169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2059..2078)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2113..2132
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2280..2296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2280..2296)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2293..2315)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2304..2320
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2328..2334,2335..2352,2353..2358))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2373..2394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2511..2528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2682..3270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2762..2781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3441..4232,4233..4301))
                     /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     4064..4083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4302..4406)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4474..4492
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4530..4552)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(4652..4663,1..8)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
       61 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
      121 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
      181 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg
      241 agctcggtac ccgggggcgc gccctaccgg gtaggggagg cgcttttccc aaggcagtct
      301 ggagcatgcg ctttagcggc cccgctgggc acttggcgct acacaagtgg cctctggcct
      361 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc
      421 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg
      481 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc
      541 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc
      601 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc
      661 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag
      721 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcatccat
      781 ctagatcctt aattaagtct agagtcgagg aactgaaaaa ccagaaagtt aactggtaag
      841 tttagtcttt ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac
      901 tgctcctcag tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct
      961 aaaagctgcg gaattgtacc cgcggccgat ccgacatcat gggaaaatca aaagaaatca
     1021 gccaagacct cagaaaaaaa attgtagacc tccacaagtc tggttcatcc ttgggagcaa
     1081 tttccaaacg cctggccgta ccacgttcat ctgtacaaac aatagtacgc aagtataaac
     1141 accatgggac cacgcagccg tcataccgct caggaaggag acgcgttctg tctcctagag
     1201 atgaacgtac tttggtgcga aaagtgcaaa tcaatcccag aacagccgca aaggaccttg
     1261 tgaagatgct ggaggaaaca ggtacaaaag tatctatatc cacagtaaaa cgagtcctat
     1321 atcgacataa cctgaaaggc cgctcagcaa ggaagaagcc actgctccaa aaccgacata
     1381 agaaagccag actacggttt gcaagagcac atggggacaa agatcgtact ttttggagaa
     1441 atgtcctctg gtctgatgaa acaaaaatag aactgtttgg ccataatgac catcgttatg
     1501 tttggaggaa gaagggggag gcttgcaagc cgaagaacac catcccaacc gtgaagcacg
     1561 ggggtggcag catcatgttg tgggggtgct ttgctgcagg agggactggt gcacttcaca
     1621 aaatagatgg catcatgagg aaggaaaatt atgtggatat attgaagcaa catctcaaga
     1681 catcagtcag gaagttaaag cttggtcgca aatgggtctt ccaaatggac aatgacccca
     1741 agcatacttc caaagttgtg gcaaaatggc ttaaggacaa caaagtcaag gtattggagt
     1801 ggccatcaca aagccctgac ctcaatccta tagaaaattt gtgggcagaa ctgaaaaagc
     1861 gtgtgcgagc aaggaggcct acaaacctga ctcagttaca ccagctctgt caggaggaat
     1921 gggccaaaat tcacccaact tattgtggga agcttgtgga aggctacccg aaacgtttga
     1981 cccaagttaa acaatttaaa ggcaatgcta ccaaatacta gaattggccg cggggatcca
     2041 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2101 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2161 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2221 gaggtttttt cggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     2281 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2341 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2401 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2461 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2521 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2581 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2641 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2701 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2761 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2821 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2881 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2941 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3001 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3061 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3121 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3181 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3481 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3541 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3841 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4321 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4381 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4441 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4501 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4561 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4621 gttggcgggt gtcggggctg gcttaactat gcggcatcag agc
//
