LOCUS       sequence-173569-        6042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-tagged PH(K14A) domain of human Akt1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6042)
  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 6042)
  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: pmCherry-PH1(K14A)
FEATURES             Location/Qualifiers
     source          1..6042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        783..1086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1087..1290
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1241..1261
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1329..1338
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1335..2042
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1480..1498)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1748..1767
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2064..3503
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHARGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     polyA_signal    3624..3745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3661..3680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3715..3734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3752..4207)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3889..3910)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4101..4120
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4234..4338
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4363..4383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4407..4617
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4429..4625
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4476..4611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4660..5454
                     /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             4660..5454
                     /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(4714..4733)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5324..5343
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5642..5661)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5686..5733
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
      721 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      781 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      841 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      901 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      961 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1021 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1081 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     1141 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     1201 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     1261 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
     1321 taccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     1381 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1441 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1501 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1561 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1621 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1681 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     1741 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     1801 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     1861 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     1921 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     1981 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2041 agtccggact cagatctcga gctatgagcg acgtggctat tgtgaaggag ggttggctgc
     2101 acgcacgagg ggagtacatc aagacctggc ggccacgcta cttcctcctc aagaatgatg
     2161 gcaccttcat tggctacaag gagcggccgc aggatgtgga ccaacgtgag gctcccctca
     2221 acaacttctc tgtggcgcag tgccagctga tgaagacgga gcggccccgg cccaacacct
     2281 tcatcatccg ctgcctgcag tggaccactg tcatcgaacg caccttccat gtggagactc
     2341 ctgaggagcg ggaggagtgg acaaccgcca tccagactgt ggctgacggc ctcaagaagc
     2401 aggaggagga ggagatggac ttccggtcgg gctcacccag tgacaactca ggggctgaag
     2461 agatggaggt gtccctggcc aagcccaagc accgcgtgac catgaacgag tttgagtacc
     2521 tgaagctgct gggcaagggc actttcggca aggtgatcct ggtgaaggag aaggccacag
     2581 gccgctacta cgccatgaag atcctcaaga aggaagtcat cgtggccaag gacgaggtgg
     2641 cccacacact caccgagaac cgcgtcctgc agaactccag gcaccccttc ctcacagccc
     2701 tgaagtactc tttccagacc cacgaccgcc tctgctttgt catggagtac gccaacgggg
     2761 gcgagctgtt cttccacctg tcccgggagc gtgtgttctc cgaggaccgg gcccgcttct
     2821 atggcgctga gattgtgtca gccctggact acctgcactc ggagaagaac gtggtgtacc
     2881 gggacctcaa gctggagaac ctcatgctgg acaaggacgg gcacattaag atcacagact
     2941 tcgggctgtg caaggagggg atcaaggacg gtgccaccat gaagaccttt tgcggcacac
     3001 ctgagtacct ggcccccgag gtgctggagg acaatgacta cggccgtgca gtggactggt
     3061 gggggctggg cgtggtcatg tacgagatga tgtgcggtcg cctgcccttc tacaaccagg
     3121 accatgagaa gctttttgag ctcatcctca tggaggagat ccgcttcccg cgcacgcttg
     3181 gtcccgaggc caagtccttg ctttcagggc tgctcaagaa ggaccccaag cagaggcttg
     3241 gcgggggctc cgaggacgcc aaggagatca tgcagcatcg cttctttgcc ggtatcgtgt
     3301 ggcagcacgt gtacgagaag aagctcagcc cacccttcaa gccccaggtc acgtcggaga
     3361 ctgacaccag gtattttgat gaggagttca cggcccagat gatcaccatc acaccacctg
     3421 accaagatga cagcatggag tgtgtggaca gcgagcgcag gccccacttc ccccagttct
     3481 cctactcggc cagcggcacg gcctgatcta gataactgat cataatcagc cataccacat
     3541 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3601 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3661 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3721 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     3781 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3841 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3901 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3961 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4021 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4081 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4141 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4201 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4261 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4321 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4621 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4681 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4741 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4801 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4861 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4921 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4981 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5041 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5101 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5161 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5221 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5281 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5341 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5401 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5461 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5521 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5581 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5641 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5701 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5761 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5821 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5881 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     5941 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6001 cttaacgtga gttttcgttc cactgagcgt cagaccccgt ag
//
