LOCUS       sequence-228277-        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"
     primer_bind     129..151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     143..160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     144..160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        187..490
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        491..694
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     645..665
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     691..715
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          832..928
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      984..993
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             990..2939
                     /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    2992..3126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3016..3035)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3070..3089
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3237..3253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3237..3253)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3250..3272)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3261..3277
                     /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(3285..3291,3292..3309,3310..3315))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3330..3351
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3468..3485)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3639..4227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3719..3738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4398..5189,5190..5258))
                     /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     5021..5040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5259..5363)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5431..5449
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5487..5509)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5609..5628
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
       61 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      121 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc ttgcatgcct
      181 gcaggtcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      241 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      301 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      361 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      421 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      481 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      541 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      601 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      661 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
      721 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      781 gactctagag gatccggtac tcgaggaact gaaaaaccag aaagttaact ggtaagttta
      841 gtctttttgt cttttatttc aggtcccgga tccggtggtg gtgcaaatca aagaactgct
      901 cctcagtgga tgttgccttt acttctaggc ctgtacggaa gtgttacttc tgctctaaaa
      961 gctgcggaat tgtacccgcg gccgccacca tggaggaagt atgtgattca tcagcagctg
     1021 cgagcagcac agtccaaaat cagccacagg atcaagagca cccgtggccg tatcttcgcg
     1081 aattcttttc tttaagtggt gtaaataaag attcattcaa gatgaaatgt gtcctctgtc
     1141 tcccgcttaa taaagaaata tcggccttca aaagttcgcc atcaaaccta aggaagcata
     1201 ttgagagaat gcacccaaat tacctcaaaa actactctaa attgacagca cagaagagaa
     1261 agatcgggac ctccacccat gcttccagca gtaagcaact gaaagttgac tcagttttcc
     1321 cagtcaaaca tgtgtctcca gtcactgtga acaaagctat attaaggtac atcattcaag
     1381 gacttcatcc tttcagcact gttgatctgc catcatttaa agagctgatt agtacactgc
     1441 agcctggcat ttctgtcatt acaaggccta ctttacgctc caagatagct gaagctgctc
     1501 tgatcatgaa acagaaagtg actgctgcca tgagtgaagt tgaatggatt gcaaccacaa
     1561 cggattgttg gactgcacgt agaaagtcat tcattggtgt aactgctcac tggatcaacc
     1621 ctggaagtct tgaaagacat tccgctgcac ttgcctgcaa aagattaatg ggctctcata
     1681 cttttgaggt actggccagt gccatgaatg atatccactc agagtatgaa atacgtgaca
     1741 aggttgtttg cacaaccaca gacagtggtt ccaactttat gaaggctttc agagtttttg
     1801 gtgtggaaaa caatgatatc gagactgagg caagaaggtg tgaaagtgat gacactgatt
     1861 ctgaaggctg tggtgaggga agtgatggtg tggaattcca agatgcctca cgagtcctgg
     1921 accaagacga tggcttcgaa ttccagctac caaaacatca aaagtgtgcc tgtcacttac
     1981 ttaacctagt ctcaagcgtt gatgcccaaa aagctctctc aaatgagcac tacaagaaac
     2041 tctacagatc tgtctttggc aaatgccaag ctttatggaa taaaagcagc cgatcggctc
     2101 tagcagctga agctgttgaa tcagaaagcc ggcttcagct tttaaggcca aaccaaacgc
     2161 ggtggaattc aacttttatg gctgttgaca gaattcttca aatttgcaaa gaagcaggag
     2221 aaggcgcact tcggaatata tgcacctctc ttgaggttcc aatgtttaat ccagcagaaa
     2281 tgctgttctt gacagagtgg gccaacacaa tgcgtccagt tgcaaaagta ctcgacatct
     2341 tgcaagcgga aacgaataca cagctggggt ggctgctgcc tagtgtccat cagttaagct
     2401 tgaaacttca gcgactccac cattctctca ggtactgtga cccacttgtg gatgccctac
     2461 aacaaggaat ccaaacacga ttcaagcata tgtttgaaga tcctgagatc atagcagctg
     2521 ccatccttct ccctaaattt cggacctctt ggacaaatga tgaaaccatc ataaaacgag
     2581 gcatggacta catcagagtg catctggagc ctttggacca caagaaggaa ttggccaaca
     2641 gttcatctga tgatgaagat tttttcgctt ctttgaaacc gacaacacat gaagccagca
     2701 aagagttgga tggatatctg gcctgtgttt cagacaccag ggagtctctg ctcacgtttc
     2761 ctgctatttg cagcctctct atcaagacta atacacctct tcccgcatcg gctgcctgtg
     2821 agaggctttt cagcactgca ggattgcttt tcagccccaa aagagctagg cttgacacta
     2881 acaattttga gaatcagctt ctactgaagt taaatctgag gttttacaac tttgagtaga
     2941 ctagtctgaa gggcgaattc tgcagatatc catcacactg gcggccgcgg ggatccagac
     3001 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3061 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3121 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     3181 gttttttcgg atcctctaga gtcgacctgc aggcatgcaa gcttggcgta atcatggtca
     3241 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3301 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3361 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3421 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3481 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3541 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3601 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3661 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3721 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3781 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3841 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3901 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3961 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4021 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4081 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4141 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4201 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4261 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4321 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4381 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4441 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4501 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4561 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4621 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4681 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4741 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4801 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4861 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4921 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4981 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5041 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5101 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5161 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5221 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5281 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5341 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5401 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5461 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5521 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5581 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5641 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat cag
//
