LOCUS       sequence-35858-e        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5693)
  AUTHORS   Balciunas D, Wangensteen KJ, Wilber A, Bell J, Geurts A, Sivasubbu 
            S, Wang X, Hackett PB, Largaespada DA, McIvor RS, Ekker SC
  TITLE     Harnessing a high cargo-capacity transposon for genetic applications
            in vertebrates.
  JOURNAL   PLoS Genet. 2006 Nov 10;2(11):e169. Epub 2006 Aug 28.
  PUBMED    17096595
REFERENCE   2  (bases 1 to 5693)
  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"; journalName: "PLoS 
            Genet."; date: "2006-11-10"; volume: "2(11)"; pages: "e169. Epub 
            2006 Aug 28"
COMMENT     Sequence Label: pCMV-Tol2
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        26..329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        330..533
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     484..504
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     530..554
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          671..767
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      823..832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             829..2778
                     /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"
     polyA_signal    2831..2965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2855..2874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2909..2928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3076..3092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3076..3092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3089..3111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3100..3116
                     /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(3124..3130,3131..3148,3149..3154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3169..3190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3307..3324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3478..4066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3558..3577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4237..5028,5029..5097))
                     /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     4860..4879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5098..5202)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5270..5288
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5326..5348)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5448..5467
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5661..5683
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5675..5692
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5676..5692
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattcgagct tgcatgcctg caggtcgtta cataacttac ggtaaatggc ccgcctggct
       61 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      121 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      181 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      241 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      301 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      361 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      421 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      481 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      541 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      601 gggaccgatc cagcctccgg actctagagg atccggtact cgaggaactg aaaaaccaga
      661 aagttaactg gtaagtttag tctttttgtc ttttatttca ggtcccggat ccggtggtgg
      721 tgcaaatcaa agaactgctc ctcagtggat gttgccttta cttctaggcc tgtacggaag
      781 tgttacttct gctctaaaag ctgcggaatt gtacccgcgg ccgccaccat ggaggaagta
      841 tgtgattcat cagcagctgc gagcagcaca gtccaaaatc agccacagga tcaagagcac
      901 ccgtggccgt atcttcgcga attcttttct ttaagtggtg taaataaaga ttcattcaag
      961 atgaaatgtg tcctctgtct cccgcttaat aaagaaatat cggccttcaa aagttcgcca
     1021 tcaaacctaa ggaagcatat tgagagaatg cacccaaatt acctcaaaaa ctactctaaa
     1081 ttgacagcac agaagagaaa gatcgggacc tccacccatg cttccagcag taagcaactg
     1141 aaagttgact cagttttccc agtcaaacat gtgtctccag tcactgtgaa caaagctata
     1201 ttaaggtaca tcattcaagg acttcatcct ttcagcactg ttgatctgcc atcatttaaa
     1261 gagctgatta gtacactgca gcctggcatt tctgtcatta caaggcctac tttacgctcc
     1321 aagatagctg aagctgctct gatcatgaaa cagaaagtga ctgctgccat gagtgaagtt
     1381 gaatggattg caaccacaac ggattgttgg actgcacgta gaaagtcatt cattggtgta
     1441 actgctcact ggatcaaccc tggaagtctt gaaagacatt ccgctgcact tgcctgcaaa
     1501 agattaatgg gctctcatac ttttgaggta ctggccagtg ccatgaatga tatccactca
     1561 gagtatgaaa tacgtgacaa ggttgtttgc acaaccacag acagtggttc caactttatg
     1621 aaggctttca gagtttttgg tgtggaaaac aatgatatcg agactgaggc aagaaggtgt
     1681 gaaagtgatg acactgattc tgaaggctgt ggtgagggaa gtgatggtgt ggaattccaa
     1741 gatgcctcac gagtcctgga ccaagacgat ggcttcgaat tccagctacc aaaacatcaa
     1801 aagtgtgcct gtcacttact taacctagtc tcaagcgttg atgcccaaaa agctctctca
     1861 aatgaacact acaagaaact ctacagatct gtctttggca aatgccaagc tttatggaat
     1921 aaaagcagcc gatcggctct agcagctgaa gctgttgaat cagaaagccg gcttcagctt
     1981 ttaaggccaa accaaacgcg gtggaattca acttttatgg ctgttgacag aattcttcaa
     2041 atttgcaaag aagcaggaga aggcgcactt cggaatatat gcacctctct tgaggttcca
     2101 atgtttaatc cagcagaaat gctgttcttg acagagtggg ccaacacaat gcgtccagtt
     2161 gcaaaagtac tcgacatctt gcaagcggaa acgaatacac agctggggtg gctgctgcct
     2221 agtgtccatc agttaagctt gaaacttcag cgactccacc attctctcag gtactgtgac
     2281 ccacttgtgg atgccctaca acaaggaatc caaacacgat tcaagcatat gtttgaagat
     2341 cctgagatca tagcagctgc catccttctc cctaaatttc ggacctcttg gacaaatgat
     2401 gaaaccatca taaaacgagg catggactac atcagagtgc atctggagcc tttggaccac
     2461 aagaaggaat tggccaacag ttcatctgat gatgaagatt ttttcgcttc tttgaaaccg
     2521 acaacacatg aagccagcaa agagttggat ggatatctgg cctgtgtttc agacaccagg
     2581 gagtctctgc tcacgtttcc tgctatttgc agcctctcta tcaagactaa tacacctctt
     2641 cccgcatcgg ctgcctgtga gaggcttttc agcactgcag gattgctttt cagccccaaa
     2701 agagctaggc ttgacactaa caattttgag aatcagcttc tactgaagtt aaatctgagg
     2761 ttttacaact ttgagtagac tagtctgaag ggcgaattct gcagatatcc atcacactgg
     2821 cggccgcggg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2881 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2941 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3001 tcagggggag gtgtgggagg ttttttcgga tcctctagag tcgacctgca ggcatgcaag
     3061 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3121 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3181 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3241 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3301 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3901 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4141 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4201 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4261 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4321 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4381 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4441 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4501 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4561 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4621 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4681 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4741 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4801 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4861 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4921 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4981 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5041 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5101 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5161 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5221 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5281 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5341 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5401 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5461 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5521 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5581 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5641 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtg
//
