LOCUS       sequence-222606-        6992 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 6992)
  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 6992)
  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..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1821..3770
                     /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(4014..4033)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4079..4134
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4133..4152)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4495..4511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4495..4511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4508..4530)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4519..4535
                     /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(4543..4549,4550..4567,4568..4573))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4588..4609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4668..4863
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4714..4849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4869..5003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4906..4925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4960..4979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5071..5088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5242..5830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5322..5341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6001..6792,6793..6861))
                     /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     6624..6643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6862..6966)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gcggggsggg gcgrggggsg
      541 gggsggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgaggt
     1741 cgagtcacat ctattaccac aatgcacagc accttgacct ggaaattagg gaaattataa
     1801 cagtcaatca gtggaagaaa atggaggaag tatgtgattc atcagcagct gcgagcagca
     1861 cagtccaaaa tcagccacag gatcaagagc acccgtggcc gtatcttcgc gaattctttt
     1921 ctttaagtgg tgtaaataaa gattcattca agatgaaatg tgtcctctgt ctcccgctta
     1981 ataaagaaat atcggccttc aaaagttcgc catcaaacct aaggaagcat attgagagaa
     2041 tgcacccaaa ttacctcaaa aactactcta aattgacagc acagaagaga aagatcggga
     2101 cctccaccca tgcttccagc agtaagcaac tgaaagttga ctcagttttc ccagtcaaac
     2161 atgtgtctcc agtcactgtg aacaaagcta tattaaggta catcattcaa ggacttcatc
     2221 ctttcagcac tgttgatctg ccatcattta aagagctgat tagtacactg cagcctggca
     2281 tttctgtcat tacaaggcct actttacgct ccaagatagc tgaagctgct ctgatcatga
     2341 aacagaaagt gactgctgcc atgagtgaag ttgaatggat tgcaaccaca acggattgtt
     2401 ggactgcacg tagaaagtca ttcattggtg taactgctca ctggatcaac cctggaagtc
     2461 ttgaaagaca ttccgctgca cttgcctgca aaagattaat gggctctcat acttttgagg
     2521 tactggccag tgccatgaat gatatccact cagagtatga aatacgtgac aaggttgttt
     2581 gcacaaccac agacagtggt tccaacttta tgaaggcttt cagagttttt ggtgtggaaa
     2641 acaatgatat cgagactgag gcaagaaggt gtgaaagtga tgacactgat tctgaaggct
     2701 gtggtgaggg aagtgatggt gtggaattcc aagatgcctc acgagtcctg gaccaagacg
     2761 atggcttcga attccagcta ccaaaacatc aaaagtgtgc ctgtcactta cttaacctag
     2821 tctcaagcgt tgatgcccaa aaagctctct caaatgaaca ctacaagaaa ctctacagat
     2881 ctgtctttgg caaatgccaa gctttatgga ataaaagcag ccgatcggct ctagcagctg
     2941 aagctgttga atcagaaagc cggcttcagc ttttaaggcc aaaccaaacg cggtggaatt
     3001 caacttttat ggctgttgac agaattcttc aaatttgcaa agaagcagga gaaggcgcac
     3061 ttcggaatat atgcacctct cttgaggttc caatgtttaa tccagcagaa atgctgttct
     3121 tgacagagtg ggccaacaca atgcgtccag ttgcaaaagt actcgacatc ttgcaagcgg
     3181 aaacgaatac acagctgggg tggctgctgc ctagtgtcca tcagttaagc ttgaaacttc
     3241 agcgactcca ccattctctc aggtactgtg acccacttgt ggatgcccta caacaaggaa
     3301 tccaaacacg attcaagcat atgtttgaag atcctgagat catagcagct gccatccttc
     3361 tccctaaatt tcggacctct tggacaaatg atgaaaccat cataaaacga ggcatggact
     3421 acatcagagt gcatctggag cctttggacc acaagaagga attggccaac agttcatctg
     3481 atgatgaaga ttttttcgct tctttgaaac cgacaacaca tgaagccagc aaagagttgg
     3541 atggatatct ggcctgtgtt tcagacacca gggagtctct gctcacgttt cctgctattt
     3601 gcagcctctc tatcaagact aatacacctc ttcccgcatc ggctgcctgt gagaggcttt
     3661 tcagcactgc aggattgctt ttcagcccca aaagagctag gcttgacact aacaattttg
     3721 agaatcagct tctactgaag ttaaatctga ggttttacaa ctttgagtag cgtgtactgg
     3781 cattagattg tctgtcttat agtttgataa ttaaatacaa acagttctaa agcaggataa
     3841 aaccttgtat gcatttcatt taatgttttt tgagattaaa agcttaaaca agaaaaaaaa
     3901 aaaaaaaaaa aaaaaaaagt actagtcgag gaattcactc ctcaggtgca ggctgcctat
     3961 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     4021 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     4081 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     4141 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     4201 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     4261 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     4321 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     4381 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     4441 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     4501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     4681 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4741 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4801 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4861 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4921 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4981 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     5041 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5101 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6961 tccgcgcaca tttccccgaa aagtgccacc tg
//
