LOCUS       sequence-72751-e        4477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cDNA of Akt1 with myristyolation signal and point mutation from K to
            M at position 179 in cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4477)
  AUTHORS   Aoki M, Batista O, Bellacosa A, Tsichlis P, Vogt PK
  TITLE     The akt kinase: molecular determinants of oncogenicity.
  JOURNAL   Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14950-5.
  PUBMED    9843996
REFERENCE   2  (bases 1 to 4477)
  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: "Proc Natl 
            Acad Sci U S A."; date: "1998-12-8"; volume: "95(25)"; pages: 
            "14950-5"
COMMENT     Sequence Label: pBSFI-myr-akt-K179M
FEATURES             Location/Qualifiers
     source          1..4477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..83
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             1536..1562
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1631..1650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1632..1650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1660..1677)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1660..1676)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1669..1691)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1817..2272)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1954..1975)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2166..2185
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2299..2403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2404..2472,2473..3264)
                     /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     complement(2622..2641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3435..4023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3924..3943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4177..4194
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4311..4332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4347..4352,4353..4370,4371..4377)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4385..4401
                     /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)."
     primer_bind     4390..4412
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4409..4425
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4409..4425
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4446..4464
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agctggccat tacggccgcg gccgcctcta gaggatctgg taccacgcgt atcgataagc
       61 ttatggggag cagcaagagc aagcccaagt ctagaatgaa cgacgtagcc attgtgaagg
      121 agggctggct gcacaaacga ggggaatata ttaaaacctg gcggccacgc tacttcctcc
      181 tcaagaacga tggcaccttt attggctaca aggaacggcc tcaggatgtg gatcagcgag
      241 agtccccact caacaacttc tcagtggcac aatgccagct gatgaagaca gagcggccaa
      301 ggcccaacac ctttatcatc cgctgcctgc agtggaccac agtcattgag cgcaccttcc
      361 atgtggaaac gcctgaggag cgggaagaat gggccaccgc cattcagact gtggccgatg
      421 gactcaagag gcaggaagaa gagacgatgg acttccgatc aggctcaccc agtgacaact
      481 caggggctga agagatggag gtgtccctgg ccaagcccaa gcaccgtgtg accatgaacg
      541 agtttgagta cctgaaacta ctgggcaagg gcacctttgg gaaagtgatt ctggtgaaag
      601 agaaggccac aggccgctac tatgccatga tgatcctcaa gaaggaggtc atcgtcgcca
      661 aggatgaggt tgcccacacg cttactgaga accgtgtcct gcagaactct aggcatccct
      721 tccttacggc cctcaagtac tcattccaga cccacgaccg cctctgcttt gtcatggagt
      781 atgccaacgg gggcgagctc ttcttccacc tgtctcgaga gcgcgtgttc tccgaggacc
      841 gggcccgctt ctatggtgcg gagattgtgt ctgccctgga ctacttgcac tccgagaaga
      901 acgtggtgta ccgggacctg aagctggaga acctcatgct ggacaaggac gggcacatca
      961 agataacgga cttcgggctg tgcaaggagg ggatcaagga tggtgccact atgaagacat
     1021 tctgcggaac gccggagtac ctggcccctg aggtgctgga ggacaacgac tacggccgtg
     1081 cagtggactg gtgggggctg ggcgtggtca tgtatgagat gatgtgtggc cgcctgccct
     1141 tctacaacca ggaccacgag aagctgttcg agctgatcct catggaggag atccgcttcc
     1201 cgcgcacact cggccctgag gccaagtccc tgctctccgg gctgctcaag aaggacccta
     1261 cacagaggct cggtgggggc tctgaggatg ccaaggagat catgcagcac cggttctttg
     1321 ccaacatcgt gtggcaggat gtgtatgaga agaagctgag cccacctttc aagccccagg
     1381 tcacctctga gactgacacc aggtatttcg atgaggagtt cacagctcag atgatcacca
     1441 tcacgccgcc tgatcaagat gacagcatgg agtgtgtgga cagtgagcgg aggccgcact
     1501 tcccccagtt ctcctactca gccagtggca cagcctaccc atacgatgtt ccagattacg
     1561 cttctagatg aattcacgcg taagcttatc gatgtcgacg ggcccggccg cctcggccgt
     1621 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     1681 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1741 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1801 cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     1861 tgttaaatca gctcattttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1921 aaaagataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     1981 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2041 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2101 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2161 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2221 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     2281 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2401 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2461 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2521 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2581 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2641 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2701 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2761 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2821 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2881 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2941 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3001 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3061 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3121 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3181 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3241 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3301 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3541 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4141 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4201 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4261 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     4321 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     4381 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     4441 ctcgaaatta accctcacta aagggaacaa aagctgg
//
