LOCUS       sequence-216865-        6982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6982)
  AUTHORS   Kawauchi D, Ogg RJ, Liu L, Shih DJH, Finkelstein D, Murphy BL, Rehg 
            JE, Korshunov A, Calabrese C, Zindy F, Phoenix T, Kawaguchi Y, 
            Gronych J, Gilbertson RJ, Lichter P, Gajjar A, Kool M, Northcott PA,
            Pfister SM, Roussel MF
  TITLE     Novel MYC-driven medulloblastoma models from multiple embryonic 
            cerebellar cells.
  JOURNAL   Oncogene. 2017 Sep 14;36(37):5231-5242. doi: 10.1038/onc.2017.110. 
            Epub 2017 May 15.
  PUBMED    28504719
REFERENCE   2  (bases 1 to 6982)
  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/onc.2017.110"; journalName: "Oncogene"; date: "2017-09-14- 
            14"; volume: "36"; issue: "37"; pages: "5231-5242"
COMMENT     Sequence Label: pT2K-CAGGS-T2TP
FEATURES             Location/Qualifiers
     source          1..6982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1813..3762
                     /codon_start=1
                     /product="transposase of the Tol2 transposon from the 
                     medaka fish Oryzias latipes (Kawakami, 2007)"
                     /label=Tol2 transposase
                     /translation="MEEVCDSSAAASSTVQNQPQDQEHPWPYLREFFSLSGVNKDSFKM
                     KCVLCLPLNKEISAFKSSPSNLRKHIERMHPNYLKNYSKLTAQKRKIGTSTHASSSKQL
                     KVDSVFPVKHVSPVTVNKAILRYIIQGLHPFSTVDLPSFKELISTLQPGISVITRPTLR
                     SKIAEAALIMKQKVTAAMSEVEWIATTTDCWTARRKSFIGVTAHWINPGSLERHSAALA
                     CKRLMGSHTFEVLASAMNDIHSEYEIRDKVVCTTTDSGSNFMKAFRVFGVENNDIETEA
                     RRCESDDTDSEGCGEGSDGVEFQDASRVLDQDDGFEFQLPKHQKCACHLLNLVSSVDAQ
                     KALSNEHYKKLYRSVFGKCQALWNKSSRSALAAEAVESESRLQLLRPNQTRWNSTFMAV
                     DRILQICKEAGEGALRNICTSLEVPMFNPAEMLFLTEWANTMRPVAKVLDILQAETNTQ
                     LGWLLPSVHQLSLKLQRLHHSLRYCDPLVDALQQGIQTRFKHMFEDPEIIAAAILLPKF
                     RTSWTNDETIIKRGMDYIRVHLEPLDHKKELANSSSDDEDFFASLKPTTHEASKELDGY
                     LACVSDTRESLLTFPAICSLSIKTNTPLPASAACERLFSTAGLLFSPKRARLDTNNFEN
                     QLLLKLNLRFYNFE"
     primer_bind     complement(4006..4025)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4071..4126
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4485..4501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4485..4501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4498..4520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4509..4525
                     /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(4533..4539,4540..4557,4558..4563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4578..4599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4658..4853
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4704..4839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4859..4993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4896..4915)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4950..4969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5061..5078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5232..5820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5312..5331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5991..6782,6783..6851))
                     /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     6614..6633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6852..6956)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggaa cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgag gtcgagtcac
     1741 atctattacc acaatgcaca gcaccttgac ctggaaatta gggaaattat aacagtcaat
     1801 cagtggaaga aaatggagga agtatgtgat tcatcagcag ctgcgagcag cacagtccaa
     1861 aatcagccac aggatcaaga gcacccgtgg ccgtatcttc gcgaattctt ttctttaagt
     1921 ggtgtaaata aagattcatt caagatgaaa tgtgtcctct gtctcccgct taataaagaa
     1981 atatcggcct tcaaaagttc gccatcaaac ctaaggaagc atattgagag aatgcaccca
     2041 aattacctca aaaactactc taaattgaca gcacagaaga gaaagatcgg gacctccacc
     2101 catgcttcca gcagtaagca actgaaagtt gactcagttt tcccagtcaa acatgtgtct
     2161 ccagtcactg tgaacaaagc tatattaagg tacatcattc aaggacttca tcctttcagc
     2221 actgttgatc tgccatcatt taaagagctg attagtacac tgcagcctgg catttctgtc
     2281 attacaaggc ctactttacg ctccaagata gctgaagctg ctctgatcat gaaacagaaa
     2341 gtgactgctg ccatgagtga agttgaatgg attgcaacca caacggattg ttggactgca
     2401 cgtagaaagt cattcattgg tgtaactgct cactggatca accctggaag tcttgaaaga
     2461 cattccgctg cacttgcctg caaaagatta atgggctctc atacttttga ggtactggcc
     2521 agtgccatga atgatatcca ctcagagtat gaaatacgtg acaaggttgt ttgcacaacc
     2581 acagacagtg gttccaactt tatgaaggct ttcagagttt ttggtgtgga aaacaatgat
     2641 atcgagactg aggcaagaag gtgtgaaagt gatgacactg attctgaagg ctgtggtgag
     2701 ggaagtgatg gtgtggaatt ccaagatgcc tcacgagtcc tggaccaaga cgatggcttc
     2761 gaattccagc taccaaaaca tcaaaagtgt gcctgtcact tacttaacct agtctcaagc
     2821 gttgatgccc aaaaagctct ctcaaatgaa cactacaaga aactctacag atctgtcttt
     2881 ggcaaatgcc aagctttatg gaataaaagc agccgatcgg ctctagcagc tgaagctgtt
     2941 gaatcagaaa gccggcttca gcttttaagg ccaaaccaaa cgcggtggaa ttcaactttt
     3001 atggctgttg acagaattct tcaaatttgc aaagaagcag gagaaggcgc acttcggaat
     3061 atatgcacct ctcttgaggt tccaatgttt aatccagcag aaatgctgtt cttgacagag
     3121 tgggccaaca caatgcgtcc agttgcaaaa gtactcgaca tcttgcaagc ggaaacgaat
     3181 acacagctgg ggtggctgct gcctagtgtc catcagttaa gcttgaaact tcagcgactc
     3241 caccattctc tcaggtactg tgacccactt gtggatgccc tacaacaagg aatccaaaca
     3301 cgattcaagc atatgtttga agatcctgag atcatagcag ctgccatcct tctccctaaa
     3361 tttcggacct cttggacaaa tgatgaaacc atcataaaac gaggcatgga ctacatcaga
     3421 gtgcatctgg agcctttgga ccacaagaag gaattggcca acagttcatc tgatgatgaa
     3481 gattttttcg cttctttgaa accgacaaca catgaagcca gcaaagagtt ggatggatat
     3541 ctggcctgtg tttcagacac cagggagtct ctgctcacgt ttcctgctat ttgcagcctc
     3601 tctatcaaga ctaatacacc tcttcccgca tcggctgcct gtgagaggct tttcagcact
     3661 gcaggattgc ttttcagccc caaaagagct aggcttgaca ctaacaattt tgagaatcag
     3721 cttctactga agttaaatct gaggttttac aactttgagt agcgtgtact ggcattagat
     3781 tgtctgtctt atagtttgat aattaaatac aaacagttct aaagcaggat aaaaccttgt
     3841 atgcatttca tttaatgttt tttgagatta aaagcttaaa caagaaaaaa aaaaaaaaaa
     3901 aaaaaaaaaa gtactagtcg aggaattcac tcctcaggtg caggctgcct atcagaaggt
     3961 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     4021 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     4081 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggtcata
     4141 tgggagggca aaacatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     4201 tgccatatgc tggctgccat gaacaaaggt ggctataaag aggtcatcag tatatgaaac
     4261 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     4321 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     4381 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     4441 tatgaagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     4501 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4561 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4621 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     4681 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4741 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4801 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     4861 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4921 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4981 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5401 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5641 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6961 tttccccgaa aagtgccacc tg
//
