LOCUS       sequence-72750-e        4474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cDNA of Akt1 with myristyolation signal and point mutation from T to
            A at position 308 and S to A at position 473 in cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4474)
  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 4474)
  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-T308A/S473A
FEATURES             Location/Qualifiers
     source          1..4474
                     /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(1657..1674)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1657..1673)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1666..1688)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1814..2269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1951..1972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2163..2182
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2296..2400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2401..2469,2470..3261)
                     /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(2619..2638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3432..4020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3921..3940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4174..4191
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4308..4329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4344..4349,4350..4367,4368..4374)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4382..4398
                     /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     4387..4409
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4406..4422
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4406..4422
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4443..4461
                     /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 agatcctcaa 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 atgaaggcat
     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 cgcctactca gccagtggca cagcctaccc atacgatgtt ccagattacg
     1561 cttctagatg aattcacgcg taagcttatc gatgtcgacg ggcccggccg cctcggccgt
     1621 acccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     1681 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     1741 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1801 gaatggcgaa tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     1861 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     1921 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1981 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2041 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2101 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2161 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2221 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2281 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2341 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2401 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2461 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2521 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2581 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2641 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2701 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2761 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2821 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2881 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2941 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3001 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3061 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3121 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3181 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3241 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3301 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3361 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3541 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4081 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4141 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4201 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4261 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     4321 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     4381 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     4441 gaaattaacc ctcactaaag ggaacaaaag ctgg
//
