LOCUS       sequence-167628-        5142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Source of Tol2 transposase driven by a mini-CAGS promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5142)
  AUTHORS   Clark KJ, Carlson DF, Foster LK, Kong BW, Foster DN, Fahrenkrug SC
  TITLE     Enzymatic engineering of the porcine genome with transposons and 
            recombinases.
  JOURNAL   BMC Biotechnol. 2007 Jul 17;7:42.
  PUBMED    17640337
REFERENCE   2  (bases 1 to 5142)
  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: "BMC 
            Biotechnol."; date: "2007-07-17"; volume: "7"; pages: "42"
COMMENT     Sequence Label: pKC-Tol2
FEATURES             Location/Qualifiers
     source          1..5142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        92..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..672
                     /label=chicken beta-actin promoter
     primer_bind     928..947
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             985..2934
                     /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    2979..3060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3184..3265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3375..3392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3546..4134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3626..3645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4277..5086)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5000..5019
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5078..5097
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctgagatctt aagggattac tagtttaaac ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      421 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      481 tattttgtgc agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg
      541 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      601 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      661 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      721 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      781 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      841 tggctgcgtg aaagccttga ggggctccgg gagctagagc ctctgctaac catgttcatg
      901 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
      961 ttggcaaaga attttaagct agccatggag gaagtatgtg attcatcagc agctgcgagc
     1021 agcacagtcc aaaatcagcc acaggatcaa gagcacccgt ggccgtatct tcgcgaattc
     1081 ttttctttaa gtggtgtaaa taaagattca ttcaagatga aatgtgtcct ctgtctcccg
     1141 cttaataaag aaatatcggc cttcaaaagt tcgccatcaa acctaaggaa gcatattgag
     1201 agaatgcacc caaattacct caaaaactac tctaaattga cagcacagaa gagaaagatc
     1261 gggacctcca cccatgcttc cagcagtaag caactgaaag ttgactcagt tttcccagtc
     1321 aaacatgtgt ctccagtcac tgtgaacaaa gctatattaa ggtacatcat tcaaggactt
     1381 catcctttca gcactgttga tctgccatca tttaaagagc tgattagtac actgcagcct
     1441 ggcatttctg tcattacaag gcctacttta cgctccaaga tagctgaagc tgctctgatc
     1501 atgaaacaga aagtgactgc tgccatgagt gaagttgaat ggattgcaac cacaacggat
     1561 tgttggactg cacgtagaaa gtcattcatt ggtgtaactg ctcactggat caaccctgga
     1621 agtcttgaaa gacattccgc tgcacttgcc tgcaaaagat taatgggctc tcatactttt
     1681 gaggtactgg ccagtgccat gaatgatatc cactcagagt atgaaatacg tgacaaggtt
     1741 gtttgcacaa ccacagacag tggttccaac tttatgaagg ctttcagagt ttttggtgtg
     1801 gaaaacaatg atatcgagac tgaggcaaga aggtgtgaaa gtgatgacac tgattctgaa
     1861 ggctgtggtg agggaagtga tggtgtggaa ttccaagatg cctcacgagt cctggaccaa
     1921 gacgatggct tcgaattcca gctaccaaaa catcaaaagt gtgcctgtca cttacttaac
     1981 ctagtctcaa gcgttgatgc ccaaaaagct ctctcaaatg agcactacaa gaaactctac
     2041 agatctgtct ttggcaaatg ccaagcttta tggaataaaa gcagccgatc ggctctagca
     2101 gctgaagctg ttgaatcaga aagccggctt cagcttttaa ggccaaacca aacgcggtgg
     2161 aattcaactt ttatggctgt tgacagaatt cttcaaattt gcaaagaagc aggagaaggc
     2221 gcacttcgga atatatgcac ctctcttgag gttccaatgt ttaatccagc agaaatgctg
     2281 ttcttgacag agtgggccaa cacaatgcgt ccagttgcaa aagtactcga catcttgcaa
     2341 gcggaaacga atacacagct ggggtggctg ctgcctagtg tccatcagtt aagcttgaaa
     2401 cttcagcgac tccaccattc tctcaggtac tgtgacccac ttgtggatgc cctacaacaa
     2461 ggaatccaaa cacgattcaa gcatatgttt gaagatcctg agatcatagc agctgccatc
     2521 cttctcccta aatttcggac ctcttggaca aatgatgaaa ccatcataaa acgaggcatg
     2581 gactacatca gagtgcatct ggagcctttg gaccacaaga aggaattggc caacagttca
     2641 tctgatgatg aagatttttt cgcttctttg aaaccgacaa cacatgaagc cagcaaagag
     2701 ttggatggat atctggcctg tgtttcagac accagggagt ctctgctcac gtttcctgct
     2761 atttgcagcc tctctatcaa gactaataca cctcttcccg catcggctgc ctgtgagagg
     2821 cttttcagca ctgcaggatt gcttttcagc cccaaaagag ctaggcttga cactaacaat
     2881 tttgagaatc agcttctact gaagttaaat ctgaggtttt acaactttga gtaggatcca
     2941 gaattcattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3001 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3061 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     3121 taaagcaagt aaaacctcta caaatgtggt cattgatgag tttggacaaa ccacaactag
     3181 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3241 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3301 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtcattc
     3361 agctgttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3421 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3481 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3541 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3601 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3661 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3721 ggaaccgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3781 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3841 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3901 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3961 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4021 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4081 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4141 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4201 ttggtcatga gattatcaaa aaggatcttc acctagatcc tttttgccag tgttacaacc
     4261 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     4321 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     4381 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     4441 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     4501 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     4561 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     4621 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     4681 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     4741 cacctgaatc aggatattct tctaatacct ggaatgctgt ttttccgggg atcgcagtgg
     4801 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     4861 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     4921 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     4981 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     5041 tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcataaca ccccttgtat
     5101 tactgtttat gtaagcagac agttttattg ttcatgatgc ag
//
