LOCUS       sequence-125734-        4136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human AKT1 [NM_005163.2] with stop codon (for 
            native or N-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4136)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 4136)
  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"
COMMENT     Sequence Label: R777-E001 Hs.AKT1
FEATURES             Location/Qualifiers
     source          1..4136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             186..1625
                     /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"
     protein_bind    complement(1627..1726)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2103..2894
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2987..3575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3476..3495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3729..3746
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3905..3932)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4024..4110)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
      121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaccatgag cgacgtggct attgtgaagg agggttggct gcacaaacga ggggagtaca
      241 tcaagacctg gcggccacgc tacttcctcc tcaagaatga tggcaccttc attggctaca
      301 aggagcggcc gcaggatgtg gaccaacgtg aggctcccct caacaacttc tctgtggcgc
      361 agtgccagct gatgaagacg gagcggcccc ggcccaacac cttcatcatc cgctgcctgc
      421 agtggaccac tgtcatcgaa cgcaccttcc atgtggagac tcctgaggag cgggaggagt
      481 ggacaaccgc catccagact gtggctgacg gcctcaagaa gcaggaggag gaggagatgg
      541 acttccggtc gggctcaccc agtgacaact caggggctga agagatggag gtgtccctgg
      601 ccaagcccaa gcaccgcgtg accatgaacg agtttgagta cctgaagctg ctgggcaagg
      661 gcactttcgg caaggtgatc ctggtgaagg agaaggccac aggccgctac tacgccatga
      721 agatcctcaa gaaggaagtc atcgtggcca aggacgaggt ggcccacaca ctcaccgaga
      781 accgcgtcct gcagaactcc aggcacccct tcctcacagc cctgaagtac tctttccaga
      841 cccacgaccg cctctgcttt gtcatggagt acgccaacgg gggcgagctg ttcttccacc
      901 tgtcccggga gcgtgtgttc tccgaggacc gggcccgctt ctatggcgct gagattgtgt
      961 cagccctgga ctacctgcac tcggagaaga acgtggtgta ccgggacctc aagctggaga
     1021 acctcatgct ggacaaggac gggcacatta agatcacaga cttcgggctg tgcaaggagg
     1081 ggatcaagga cggtgccacc atgaagacct tttgcggcac acctgagtac ctggcccccg
     1141 aggtgctgga ggacaatgac tacggccgtg cagtggactg gtgggggctg ggcgtggtca
     1201 tgtacgagat gatgtgcggt cgcctgccct tctacaacca ggaccatgag aagctttttg
     1261 agctcatcct catggaggag atccgcttcc cgcgcacgct tggtcccgag gccaagtcct
     1321 tgctttcagg gctgctcaag aaggacccca agcagaggct tggcgggggc tccgaggacg
     1381 ccaaggagat catgcagcat cgcttctttg ccggtatcgt gtggcagcac gtgtacgaga
     1441 agaagctcag cccacccttc aagccccagg tcacgtcgga gactgacacc aggtattttg
     1501 atgaggagtt cacggcccag atgatcacca tcacaccacc tgaccaagat gacagcatgg
     1561 agtgtgtgga cagcgagcgc aggccccact tcccccagtt ctcctactcg gccagcggca
     1621 cggcctaatc aactttcttg tacaaagttg gcattatagg aaagcattgc ttatcaattt
     1681 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgcatg caagcttaga
     1741 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     1801 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     1861 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     1921 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     1981 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     2041 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     2101 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     2161 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     2221 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2281 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2341 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2401 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2461 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2521 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2581 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2641 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     2701 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     2761 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     2821 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     2881 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     2941 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3001 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3061 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3121 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3181 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3241 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3301 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3361 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3421 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3481 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3541 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3601 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3661 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3721 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3781 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     3841 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     3901 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     3961 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     4021 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4081 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcg
//
