LOCUS       sequence-125735-        4133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human AKT1 [NM_005163.2] without stop codon 
            (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4133)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 4133)
  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-E002 Hs.AKT1-nostop
FEATURES             Location/Qualifiers
     source          1..4133
                     /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(1624..1723)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2100..2891
                     /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      2984..3572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3473..3492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3726..3743
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3902..3929)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4021..4107)
                     /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 cggcctcaac tttcttgtac aaagttggca ttataggaaa gcattgctta tcaatttgtt
     1681 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgcatgcaa gcttagatcc
     1741 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
     1801 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
     1861 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     1921 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     1981 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     2041 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     2101 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     2161 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     2221 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     2281 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     2341 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     2401 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     2461 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     2521 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     2581 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     2641 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     2701 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     2761 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     2821 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     2881 tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt
     2941 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3001 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3061 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3121 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3181 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3241 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3301 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3361 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3421 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3481 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3541 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3601 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3661 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3721 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3781 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3841 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     3901 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     3961 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     4021 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     4081 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcg
//
