LOCUS       sequence-412564-        5250 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 5250)
  AUTHORS   Wiesner SM, Decker SA, Larson JD, Ericson K, Forster C, Gallardo JL,
            Long C, Demorest ZL, Zamora EA, Low WC, SantaCruz K, Largaespada DA,
            Ohlfest JR
  TITLE     De novo induction of genetically engineered brain tumors in mice 
            using plasmid DNA.
  JOURNAL   Cancer Res. 2009 Jan 15. 69(2):431-9.
  PUBMED    19147555
REFERENCE   2  (bases 1 to 5250)
  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: "Cancer Res.
            2009 Jan 15. 69(2):431-9."
COMMENT     Sequence Label: PKT2/CLP-AKT
FEATURES             Location/Qualifiers
     source          1..5250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(296..315)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(297..315)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1263..1289
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1296..2735
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     polyA_signal    2932..3053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2943..2962)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2997..3016
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3155..3173
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3483..3500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3654..4242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3734..3753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4385..5194)
                     /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     5108..5127
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5186..5205
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctggatccag atccctatac agttgaagtc ggaagtttac atacacttaa gttggagtca
       61 ttaaaactcg tttttcaact actccacaaa tttcttgtta acaaacaata gttttggcaa
      121 gtcagttagg acatctactt tgtgcatgac acaagtcatt tttccaacaa ttgtttacag
      181 acagattatt tcacttataa ttcactgtat cacaattcca gtgggtcaga agtttacata
      241 cactaagttg actgtgcctt taaacagctt ggaaaattcc agaaaatgat ggtctcccta
      301 tagtgagtcg tattagatcc cgggattact agtttaaacc taggatgcat accggtcaag
      361 ctagcgataa ttcctgcagg agtcaatggg aaaaacccat tggagccaag tacactgact
      421 caatagggac tttccattgg gttttgccca gtacataagg tcaatagggg gtgagtcaac
      481 aggaaagtcc cattggagcc aagtacattg agtcaatagg gactttccaa tgggttttgc
      541 ccagtacata aggtcaatgg gaggtaagcc aatgggtttt tcccattact gacatgtata
      601 ctgagtcatt agggactttc caatgggttt tgcccagtac ataaggtcaa taggggtgaa
      661 tcaacaggaa agtcccattg gagccaagta cactgagtca atagggactt tccattgggt
      721 tttgcccagt acaaaaggtc aatagggggt gagtcaatgg gtttttccca ttattggcac
      781 atacataagg tcaatagggg tgactagtgg agaagagcat gcttgagggc tgagtgcccc
      841 tcagtgggca gagagcacat ggcccacagt ccctgagaag ttggggggag gggtgggcaa
      901 ttgaactggt gcctagagaa ggtggggctt gggtaaactg ggaaagtgat gtggtgtact
      961 ggctccacct ttttccccag ggtgggggag aaccatatat aagtgcagta gtctctgtga
     1021 acattcaagc ttctgccttc tccctcctgt gagtttggta agtcactgac tgtctatgcc
     1081 tgggaaaggg tgggcaggag atggggcagt gcaggaaaag tggcactatg aaccctgcag
     1141 ccctaggaat gcatctagac aattgtacta accttcttct ctttcctctc ctgacaggtt
     1201 ggtgtacagt agcttccacc atgggctgcg tttgctcgtc gaatcccgag gacgacgctg
     1261 cttacccata cgatgttcca gattacgctt cgcatatgag cgacgtggct attgtgaagg
     1321 agggttggct gcacaaacga ggggagtaca tcaagacctg gcggccacgc tacttcctcc
     1381 tcaagaatga tggcaccttc attggctaca aggagcggcc gcaggatgtg gaccaacgtg
     1441 aggctcccct caacaacttc tctgtggcgc agtgccagct gatgaagacg gagcggcccc
     1501 ggcccaacac cttcatcatc cgctgcctgc agtggaccac tgtcatcgaa cgcaccttcc
     1561 atgtggagac tcctgaggag cgggaggagt ggacaaccgc catccagact gtggctgacg
     1621 gcctcaagaa gcaggaggag gaggagatgg acttccggtc gggctcaccc agtgacaact
     1681 caggggctga agagatggag gtgtccctgg ccaagcccaa gcaccgcgtg accatgaacg
     1741 agtttgagta cctgaagctg ctgggcaagg gcactttcgg caaggtgatc ctggtgaagg
     1801 agaaggccac aggccgctac tacgccatga agatcctcaa gaaggaagtc atcgtggcca
     1861 aggacgaggt ggcccacaca ctcaccgaga accgcgtcct gcagaactcc aggcacccct
     1921 tcctcacagc cctgaagtac tctttccaga cccacgaccg cctctgcttt gtcatggagt
     1981 acgccaacgg gggcgagctg ttcttccacc tgtcccggga acgtgtgttc tccgaggacc
     2041 gggcccgctt ctatggcgct gagattgtgt cagccctgga ctacctgcac tcggagaaga
     2101 acgtggtgta ccgggacctc aagctggaga acctcatgct ggacaaggac gggcacatta
     2161 agatcacaga cttcgggctg tgcaaggagg ggatcaagga cggtgccacc atgaagacct
     2221 tttgcggcac acctgagtac ctggcccccg aggtgctgga ggacaatgac tacggccgtg
     2281 cagtggactg gtgggggctg ggcgtggtca tgtacgagat gatgtgcggt cgcctgccct
     2341 tctacaacca ggaccatgag aagctttttg agctcatcct catggaggag atccgcttcc
     2401 cgcgcacgct tggtcccgag gccaagtcct tgctttcagg gctgctcaag aaggacccca
     2461 agcagaggct tggcgggggc tccgaggacg ccaaggagat catgcagcat cgcttctttg
     2521 ccggtatcgt gtggcagcac gtgtacgaga agaagctcag cccacccttc aagccccagg
     2581 tcacgtcgga gactgacacc aggtattttg atgaggagtt cacggcccag atgatcacca
     2641 tcacaccacc tgaccaagat gacagcatgg agtgtgtgga cagcgagcgc aggccccact
     2701 tcccccagtt ctcctactcg gccagcggca cggcctgagg cggcggtgga ctgcgctgga
     2761 cgatagcttg gagggatgga gaggcggcct cgtgccatga tctgtattta atggttttta
     2821 tttctcgggt gcatttgaga gaagccacgc tgtcgaattc ctcgagaccg tacgtgcgcg
     2881 cgaatgcatc cagatctagg taccagggcc cagctagctg gccagacatg ataagataca
     2941 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3001 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3061 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     3121 agtaaaacct ctacaaatgt ggtatggaat tatcaattaa ccctcactaa agggagacca
     3181 agttaaacaa tttaaaggca atgctaccaa atactaattg agtgtatgta aacttctgac
     3241 ccactgggaa tgtgatgaaa gaaataaaag ctgaaatgaa tcattctctc tactattatt
     3301 ctgatatttc acattcttaa aataaagtgg tgatcctaac tgacctaaga cagggaattt
     3361 ttactaggat taaatgtcag gaattgtgaa aaagtgagtt taaatgtatt tggctaaggt
     3421 gtatgtaaac ttccgacttc aactgtatag ggatctggta ccatttaaat ctgttccgct
     3481 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3541 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3601 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3661 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3721 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3781 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3841 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3901 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3961 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4021 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4081 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4201 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4261 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4321 ttatcaaaaa ggatcttcac ctagatcctt tttgccagtg ttacaaccaa ttaaccaatt
     4381 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     4441 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4501 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4561 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4621 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     4681 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4741 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4801 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4861 gatattcttc taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg
     4921 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4981 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     5041 gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     5101 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     5161 gcggcctcga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     5221 aagcagacag ttttattgtt catgatgcag
//
