LOCUS       sequence-65192-e        7141 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For expression of HA and FLAG-tagged murine CAD in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7141)
  AUTHORS   Ben-Sahra I, Howell JJ, Asara JM, Manning BD
  TITLE     Stimulation of de novo pyrimidine synthesis by growth signaling 
            through mTOR and S6K1.
  JOURNAL   Science. 2013 Mar 15;339(6125):1323-8. doi: 10.1126/science.1228792.
            Epub 2013 Feb 21.
  PUBMED    23429703
REFERENCE   2  (bases 1 to 7141)
  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"; doi: 
            "10.1126/science.1228792"; journalName: "Science"; date: 
            "2013-03-15- 15"; volume: "339"; issue: "6125"; pages: "1323-8"
COMMENT     Sequence Label: pcDNA3-FLAG-HA-CAD
FEATURES             Location/Qualifiers
     source          1..7141
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     14..33
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14..32
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      73..82
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             82..105
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             106..132
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(7048..7066)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7049..7066)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7086..7103)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 ggcttatcga aattaatacg actcactata gggagaccca agcttggtac cgagctcgga
       61 tcgatatcgc cggccgccat ggactacaag gacgacgatg acaagtaccc ttatgacgtg
      121 cccgattacg ctagcggccg ctcgtctgcg tgcttgccct gtctcagctc cgacccgatg
      181 gcggccctgg tgttggagga cgggtcggtg ctgcagggcc ggccctttgg ggccgctgtg
      241 tcgactgctg gggaagtggt gtttcagacc ggcatggtcg gctacccaga ggccctcact
      301 gacccttcct acaaagctca gatcctagtg ctaacgtatc ctctcatcgg caactacggc
      361 attccctcag atgaagagga cgagttcggc ctgagcaagt ggtttgaatc ctcggagatc
      421 cacgtggcgg gacttgtagt gggagaatgc tgtcccacac ccagccactg gagtgccaac
      481 tgcaccctgc atgagtggct gcaacagcgt ggcatccccg gcctgcaagg agtggacact
      541 cgggagctga ctaagaagtt gcgggagcaa gggtctcttt tggggaagct ggtccagaaa
      601 gggacagagc cgtcagccct gccattcgtg gaccccaatg ccaggcccct ggcaccagag
      661 gtctctatta agactccacg ggtattcaat gcagggggtg cccctcggat ctgtgctctg
      721 gattgtggcc tcaagtataa tcagatccga tgtctctgcc agcttggggc tgaagttacc
      781 gtggtaccct gggaccacga gttagacagt cagaaatatg acggtctctt cctaagtaat
      841 ggacccggtg accctgcctc ttacccgggt gtggtgtcca cactgagccg agtcttgtct
      901 gaacctaacc cccgacctgt gtttggaatc tgccttggac accagctgtt ggctttagcc
      961 atcggggcca aaacttacaa gatgagatac ggaaaccgag gccacaacca gccctgttta
     1021 ctggtgggca ctgggcgctg ctttctgacg tcccagaatc atgggtttgc agtggacgca
     1081 gattcgctgc cagcaggctg ggctcctctg ttcaccaatg ccaatgactg ttccaatgaa
     1141 ggcatagtgc acgacagcct gcccttcttc agtgtccagt ttcacccaga gcaccgggcc
     1201 ggcccttcag atatggaact gctcttcgat gtatttctag aaactgtgag agaggctgca
     1261 gctgggaaca ttgggggcca gacagtaaga gaacgcttgg ctcagcgcct ctgtccccct
     1321 gagctcccca ttcctggttc tgggctcccg cctccacgaa aggttctaat cctaggctca
     1381 gggggcctct ccattggcca agctggagaa tttgactact cgggctctca ggcaattaaa
     1441 gccctaaagg aggaaaacat ccagacatta ctaatcaacc ccaacattgc cacagtgcag
     1501 acttctcagg ggctggcaga caaggtctat ttccttccca taacacttca ctacgtaacc
     1561 caggtaattc gtaatgaacg cccagatggt gtgttactga cttttggggg tcaaacagcc
     1621 ctgaactgcg gtgtggagct gaccaaagct ggggtactgg ctcgatatgg ggtccgggtc
     1681 ttgggtacac ccgtggagac cattgaactg actgaggacc gacgagcctt cgctgccagg
     1741 atggctgaga tcggagagca tgtcgccccc agtgaagcgg caaattctct tgaacaggct
     1801 caggcagctg ccgagcgact aggctaccct gtgctggtgc gagcagcctt tgccctgggt
     1861 ggtctcggtt ctggctttgc ttccaccaaa gaggaactct cagctcttgt ggctccagct
     1921 tttgcccata ccagtcaggt gctgatagac aagtctctga agggctggaa ggagattgaa
     1981 tatgaggtgg tgagagacgc ctatggcaac tgcgtaacgg tatgcaacat ggagaactta
     2041 gacccactgg gcattcatac tggtgaatcc atagtggtgg ctccgagcca gacactgaat
     2101 gacagagagt accagcttct gcgacggaca gctatcaagg tcactcagca cctgggcatt
     2161 gtcggggagt gcaatgtaca gtatgctttg aaccccgagt ctgagcagta ttacatcatt
     2221 gaagtgaatg ccaggctctc ccgaagctct gccctggcca gtaaggccac aggctatcca
     2281 ctagcctatg tggcagccaa gctagcattg ggcattcccc tgccagagct cagaaactct
     2341 gttacgggag gaacagcagc ctttgagccc agcctggact actgcgtggt aaagattcct
     2401 cgctgggacc tcagcaagtt cctacgtgtc agtacgaaga ttgggagttg catgaagagt
     2461 gttggcgagg tcatgggcat cgggcggtcc tttgaggagg ccttccaaaa ggccctgcgc
     2521 atggtcgatg agaactgcgt aggcttcgac catacagtaa agccagtcag cgacatggag
     2581 ctggagactc cgacggataa gcggatcttt gtggtggctg ccgctctgtg ggctggctac
     2641 tctgtggagc gcctctacga gctcacccgc attgattgct ggttcctgca ccggatgaag
     2701 cgcattgtga cccacgccca gttgctggaa caacaccgtg gacaagcttt accccaagac
     2761 ctgctgcacc aggccaagtg cctcggcttc tcagacaagc agattgccct ggcagtcctg
     2821 agcacagagc tggctgttcg gaagctgcgt caggaactgg ggatctgccc agcggtgaaa
     2881 cagattgaca cagttgcagc tgagtggcca gcccagacaa attacctgta cctgacatac
     2941 tggggcaaca cccatgacct cgactttcgg gcacctcacg tcctggtcct tggctctggt
     3001 gtctaccgga ttggctccag tgttgagttt gattggtgtg ctgtgggttg catccagcag
     3061 ctccggaaga tgggttataa gaccatcatg gtgaactaca acccagagac agtcagcacc
     3121 gactatgata tgtgtgaccg actctacttt gatgagatct cttttgaggt ggtgatggac
     3181 atctatgagt tagagaaccc cgaaggtgtg atcctgtcca tgggtgggca gctgcccaac
     3241 aacatggcca tggctttgca tcggcagcag tgccgagtgc tgggcacctc ccccgaagcc
     3301 atcgactcag ccgagaaccg attcaagttc tcccggctcc tagataccat cggtatcagc
     3361 cagcctcagt ggcgagagct cagtgacctt gagtctgctc gccagttctg ccacacggtc
     3421 gggtacccct gcgtagtgcg cccctcctat gtgctcagcg gtgctgctat gaatgtggcg
     3481 tacaccgacg gggacctgga gcgtttcctg agcagtgcgg ccgccgtctc taaggagcac
     3541 cccgtagtca tctccaagtt cattcaggaa gcaaaggaga tcgacgtgga cgcggtggcc
     3601 tgtgacggca tcgtgtcggc cattgccatc tctgagcacg tggagaatgc aggtgtgcat
     3661 tcaggtgatg ccacgctggt gaccccccca caagacatca ccccgaaaac tctggagcgg
     3721 atcaaagcca ttgtgcatgc tgtgggccag gagctacagg ttacaggacc cttcaatctg
     3781 cagctcattg ccaaggatga ccagctgaag gttattgaat gcaacgtgcg tgtatctcgc
     3841 tctttcccct tcgtgtccaa gacactgggt gttgatctag tagccttggc cacaaggatc
     3901 atcatggggg agaaagtaga gcccgtgggg ctcatgactg gctctggagt cgtgggagta
     3961 aaggtacctc agttctcctt ctcacgcttg gccggtgccg acgtggtact tggtgtggag
     4021 atgaccagta ctggagaggt ggccggcttc ggagagagcc gttgtgaagc ctacctcaaa
     4081 gccatgctca gcactggctt taagatccca gagaagaaca tcctgctgac tattggcagc
     4141 tacaagaaca aaagtgagct gctcccaact gtgcggttgc tggagagcct gggctacagc
     4201 ctctatgcca gcctgggtac ggcagacttc tacactgagc acggggtcaa ggtaacagct
     4261 gtggactggc actttgaaga ggcggtggat ggtgagtgcc caccacaacg gagcatcttg
     4321 gatcagctgg ctgaaaatca ctttgagcta gtgattaacc tgtccatgcg tggggctggg
     4381 ggtcggcgtc tttcctcctt cgttaccaag ggctaccgta cccggcgtct ggccgctgat
     4441 ttctctgtgc ctctcatcat tgacatcaag tgcaccaaac tctttgtgga ggccctaggc
     4501 cagatcggcc cagcccctcc tttgaaagtg cacgttgact gcatgacctc ccagaagcta
     4561 gtgcgcctgc ccggattgat tgatgtccac gtgcaccttc gggaaccagg tgggacacac
     4621 aaagaggact ttgcctcagg cacagctgct gccttggctg ggggtgtcac catggtttgt
     4681 gctatgccta atacccgacc ccccatcatt gatgctcctg ctctcgccct tgcccagaag
     4741 ctggcagagg ctggcgcccg ctgtgacttt accttattcc ttggagcctc atctgagaac
     4801 gcggggactc tgggtgctgt ggccggctct gcagcaggcc tgaagctcta cctcaacgag
     4861 accttctctg agctgcggct ggacagtgtg gcccagtgga tggagcattt tgaaacctgg
     4921 cctgcccacc tccccattgt ggcccatgca gagcggcaga gtgtcgctgc ggtcctcatg
     4981 gtggctcagc tgacccagcg tccagtgcac atatgtcatg tggctcggaa ggaagagatc
     5041 ctgttgatta aaactgcaaa ggcgcagggg ctgccagtga cctgtgaggt cgctcctcac
     5101 cacctgttcc taaaccggga agacttggag cgcctgggac ctgggaaggg agaggtccgt
     5161 cctgagcttg ggtcccgaga ggatatggaa gccctgtggg agaatatggc agtcatcgac
     5221 tgttttgcct cagaccacgc tccccatacc ttggaagaga agtgtgggcc caagcctcca
     5281 cctggcttcc cgggcctaga gaccatgttg ccgctgctgc tgacagctgt gagcgagggc
     5341 cgcctcagtc tggatgacct gctgcagcga ttgcaccaca accctcggcg gatcttccac
     5401 ctgcctctcc aggaggacac ctatgtagag gtggatctgg agcatgagtg gacagtcccc
     5461 agccacatgc ccttctctaa ggcccgctgg accccatttg aaggacagaa ggtgaagggc
     5521 acagtgcgcc gtgtggtctt gcgaggggag gtcgcctata ttgatggaca ggtgctggta
     5581 cccccgggct atggacaaga tgtacggaag tggccgcagg gggttgttcc ccagccccct
     5641 ccttcaactc ccgccaccac cgaaatcacc acgacacctg agcgaccacg ccgagtcatc
     5701 ccgggccttc ctgatggccg cttccacctg ccaccccgca tccaccgggc ctctgaccca
     5761 ggtctgccag ctgaggaacc taaagagaag ccacccagga aggtagtgga accagagctg
     5821 atgggaaccc ctgatggtcc ctgctaccct gcaccaccag tacctagaca ggcatcaccc
     5881 cagaacctag ggtcttctgg cctgctgcac ccccagatgt cgcccctact gcactcccta
     5941 gtgggccagc acatcctgtc tgtcaagcag ttcactaagg atcagatgtc tcatctgttc
     6001 aatgtggcgc acacgctacg gatgatggta cagaaggagc ggagcctcga catcctgaag
     6061 gggaaggtca tggcctccat gttctacgag gtgagcaccc gcaccagtag ctccttcgca
     6121 gcagccatgg cccggctcgg gggcgctgtt ctcagcttct cagaagctac atcctccgtc
     6181 caaaagggcg agtcccttgc tgactcggtg cagaccatga gttgctatgc tgatgtcatt
     6241 gtgctccgac accctcagcc tggagccgtg gagctggcag ccaagcactg tcgcaggcca
     6301 gtgatcaatg ctggggatgg agttggggag caccccacac aggccctgct ggacatcttc
     6361 accattcgag aagagctggg gactgtcaat ggcatgacga tcactatggt cggtgacctg
     6421 aagcatggac gcacagtgca ctccctggcc tgcctgctca cccagtaccg tgtgagccta
     6481 cgctacgtgg cacctcccag cctgcgcatg ccacccagtg tacgggactt tgtggcctcc
     6541 cgaggcacca agcaggagga gtttgagagc atcgaggagg cgctgcccga cacagacgtg
     6601 ctctacatga ctcggatcca gaaagaacga tttggctctg tccaggaata tgaagcttgc
     6661 tttggtcagt tcatcctcac tcctcacatc atgacccggg ccaagaagaa gatggtggtg
     6721 atgcacccaa tgccccgtgt caatgagata agtgtggagg tggactcgga cccccgggca
     6781 gcctacttcc gccaagctga gaacggcatg tacatccgca tggccctgct tgccaccgtg
     6841 cttggccgtt tctaggtccc agcttccttt cttgtctctt caggcccagc tgctgggcaa
     6901 ggaatcccga tgcccccccc acgggggcag cacacttagc agacaatcct ggagcataca
     6961 gatagcccaa gcacatagat gtactaaact gggaccatgc tttaggctcc agacgagagt
     7021 ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     7081 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     7141 t
//
