LOCUS       sequence-176851-        6105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TagBFP-tagged human Akt1(WT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6105)
  AUTHORS   Ebner M, Lucic I, Leonard TA, Yudushkin I
  TITLE     PI(3,4,5)P3 Engagement Restricts Akt Activity to Cellular Membranes.
  JOURNAL   Mol Cell. 2017 Feb 2;65(3):416-431.e6. doi: 
            10.1016/j.molcel.2016.12.028.
  PUBMED    28157504
REFERENCE   2  (bases 1 to 6105)
  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: 
            "10.1016/j.molcel.2016.12"; journalName: "Mol Cell"; date: 
            "2017-02-2- 2"; volume: "65"; issue: "3"; pages: "416-431.e6"
COMMENT     Sequence Label: pTagBFP-Akt1
FEATURES             Location/Qualifiers
     source          1..6105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        18..321
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        322..525
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     476..496
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             570..1268
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1290..2729
                     /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"
     polyA_signal    2850..2971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2887..2906)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2941..2960
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2978..3433)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3115..3136)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3327..3346
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3460..3564
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3566..3923
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3566..3757
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3589..3609)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3774..3909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3958..4752
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3958..4752
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4012..4031)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4622..4641
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4940..4959)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4984..5031
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5360..5948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5849..5868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
       61 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      121 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      181 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      241 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      301 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      361 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      421 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      481 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      541 cagatccgct agcgctaccg gtcgccacca tgagcgagct gattaaggag aacatgcaca
      601 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
      661 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
      721 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
      781 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
      841 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
      901 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
      961 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg
     1021 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     1081 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     1141 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     1201 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     1261 agcttaattc cggactcaga tctcgagcta tgagcgacgt ggctattgtg aaggagggtt
     1321 ggctgcacaa acgaggggag tacatcaaga cctggcggcc acgctacttc ctcctcaaga
     1381 atgatggcac cttcattggc tacaaggagc ggccgcagga tgtggaccaa cgtgaggctc
     1441 ccctcaacaa cttctctgtg gcgcagtgcc agctgatgaa gacggagcgg ccccggccca
     1501 acaccttcat catccgctgc ctgcagtgga ccactgtcat cgaacgcacc ttccatgtgg
     1561 agactcctga ggagcgggag gagtggacaa ccgccatcca gactgtggct gacggcctca
     1621 agaagcagga ggaggaggag atggacttcc ggtcgggctc acccagtgac aactcagggg
     1681 ctgaagagat ggaggtgtcc ctggccaagc ccaagcaccg cgtgaccatg aacgagtttg
     1741 agtacctgaa gctgctgggc aagggcactt tcggcaaggt gatcctggtg aaggagaagg
     1801 ccacaggccg ctactacgcc atgaagatcc tcaagaagga agtcatcgtg gccaaggacg
     1861 aggtggccca cacactcacc gagaaccgcg tcctgcagaa ctccaggcac cccttcctca
     1921 cagccctgaa gtactctttc cagacccacg accgcctctg ctttgtcatg gagtacgcca
     1981 acgggggcga gctgttcttc cacctgtccc gggagcgtgt gttctccgag gaccgggccc
     2041 gcttctatgg cgctgagatt gtgtcagccc tggactacct gcactcggag aagaacgtgg
     2101 tgtaccggga cctcaagctg gagaacctca tgctggacaa ggacgggcac attaagatca
     2161 cagacttcgg gctgtgcaag gaggggatca aggacggtgc caccatgaag accttttgcg
     2221 gcacacctga gtacctggcc cccgaggtgc tggaggacaa tgactacggc cgtgcagtgg
     2281 actggtgggg gctgggcgtg gtcatgtacg agatgatgtg cggtcgcctg cccttctaca
     2341 accaggacca tgagaagctt tttgagctca tcctcatgga ggagatccgc ttcccgcgca
     2401 cgcttggtcc cgaggccaag tccttgcttt cagggctgct caagaaggac cccaagcaga
     2461 ggcttggcgg gggctccgag gacgccaagg agatcatgca gcatcgcttc tttgccggta
     2521 tcgtgtggca gcacgtgtac gagaagaagc tcagcccacc cttcaagccc caggtcacgt
     2581 cggagactga caccaggtat tttgatgagg agttcacggc ccagatgatc accatcacac
     2641 cacctgacca agatgacagc atggagtgtg tggacagcga gcgcaggccc cacttccccc
     2701 agttctccta ctcggccagc ggcacggcct gatctagata actgatcata atcagccata
     2761 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2821 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2881 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2941 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     3001 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3061 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3121 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3181 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3241 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3301 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3361 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3421 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3481 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3541 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3601 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3841 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3901 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3961 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4021 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4081 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4141 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4201 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4261 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4321 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4381 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4441 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4501 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4561 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4621 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4681 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4741 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4801 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4861 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4921 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4981 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5041 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5101 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5161 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5221 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5281 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5341 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5401 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5461 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5521 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5581 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5641 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5701 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5761 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5821 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5881 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5941 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6001 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     6061 attagttatt aatagtaatc aattacgggg tcattagttc atagc
//
