LOCUS       sequence-167635-        5082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Passport transposon with a puromycin-thymidine kinase fusion driven 
            by a PGK promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5082)
  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 5082)
  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: pPTn2P-PTK
FEATURES             Location/Qualifiers
     source          1..5082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(255..274)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(256..274)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     632..651
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             719..1318
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPDSVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLGGDRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKAGA"
     primer_bind     1218..1238
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1325..2320
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPSVLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(2340..2357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2346..2570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2636..2717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2841..2922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3023..3041
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3315..3332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3486..4074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3566..3585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4217..5026)
                     /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     4940..4959
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5018..5037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctggatccga ttacagtgcc ttgcataagt attcaccccc tttggacttt tctacatttt
       61 gtcatgctat aaccacagat taaaatttat ttcatcgtga gtttatgtaa tggaccaaca
      121 caaaatagtg catcatttgg aagtgggggg aaatattaca tggatttcac aattatttac
      181 aaataaaaat ctgaaaagtg ttgagtgcat atgtattcac cccctttact gtgaaacccc
      241 taacaaagat ggctccctat agtgagtcgt attagatctg agatctggcc tcgcacacat
      301 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
      361 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
      421 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
      481 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
      541 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
      601 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
      661 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagcca acgccaccat
      721 ggggaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
      781 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
      841 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
      901 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
      961 cagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     1021 ttcccggctg gccgcgcagc aacagatgga aggcctcctg ggcggcgacc gacccaagga
     1081 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     1141 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     1201 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     1261 cgacgtcgag tgcccgaagg accgcgcgac ctggtgcatg acccgcaagg ccggtgccgg
     1321 atccatgccc agcgtactgc gggtttatat agacggtcct cacgggatgg ggaaaaccac
     1381 caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg tacccgagcc
     1441 gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct acaccacaca
     1501 acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa tgacaagcgc
     1561 ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc ctcatatcgg
     1621 gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct tcgaccgcca
     1681 tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca gcatgacccc
     1741 ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca caaacatcgt
     1801 gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc agcgccccgg
     1861 cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc tgcttgccaa
     1921 tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac agctttcggg
     1981 gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac gaccccatat
     2041 cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca acggcgacct
     2101 gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc ccatgcacgt
     2161 ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc tgcaacttac
     2221 ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga tctgcgacct
     2281 ggcgcgcacg tttgcccggg agatggggga ggctaactga tctagagctc gctgatcagc
     2341 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2401 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2461 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2521 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2581 ggaaagaacc aaagggcgaa ttcattgatg agtttggaca aaccacaact agaatgcagt
     2641 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2701 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2761 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtcat tgatgagttt
     2821 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2881 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2941 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3001 tacaaatgtg gtcattcaga tcaattaacc ctcactaaag ggagaccagg ttctaccaag
     3061 tattgacaca ggggggtgaa tacttatgca cccaacagat gtcaactttt ttgttctcat
     3121 tattgtttgt gtcacaataa aatttatttt gcacctccaa agtactatgc atgttttgtt
     3181 gatcaaacgg gaaaaagttt atttaagtct atttgaattc cagttagtaa cagtacataa
     3241 tgggaaaaag tccaaggggg gtgaatactt atgcaaggca ctgtaatcgg taccatttaa
     3301 atctgttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3361 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3421 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3481 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3541 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3601 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3661 ggaaccgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3721 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3781 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3841 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3901 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3961 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4021 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4081 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4141 ttggtcatga gattatcaaa aaggatcttc acctagatcc tttttgccag tgttacaacc
     4201 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     4261 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     4321 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     4381 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     4441 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     4501 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     4561 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     4621 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     4681 cacctgaatc aggatattct tctaatacct ggaatgctgt ttttccgggg atcgcagtgg
     4741 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     4801 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     4861 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     4921 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     4981 tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcataaca ccccttgtat
     5041 tactgtttat gtaagcagac agttttattg ttcatgatgc ag
//
