LOCUS       sequence-173566-        6027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses recruitable mCherry-tagged kinase domain of human 
            Akt2(WT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6027)
  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 6027)
  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: pFKBPmCherry-Akt2(KD)
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4..195
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(27..47)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      212..347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             396..1190
                     /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             396..1190
                     /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(450..469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1060..1079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1378..1397)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1422..1469
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1798..2386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2287..2306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2572..2875
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2876..3079
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3030..3050
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(3113..3115,3116..3436)
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             3461..4168
                     /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(3606..3624)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3874..3893
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5315..5436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5352..5371)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5406..5425
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5443..5898)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5580..5601)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5792..5811
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5925..6027,1..2)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
       61 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
      121 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      181 agcaaccata gccccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
      241 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
      301 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
      361 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
      421 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
      481 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
      541 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
      601 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
      661 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
      721 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
      781 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
      841 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
      901 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
      961 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1021 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1081 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1141 ccgattcaca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     1201 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     1261 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1321 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     1381 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     1441 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     1501 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     1561 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     1621 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     1681 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1741 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1801 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1861 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1921 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1981 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2041 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2101 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2161 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2221 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2281 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2341 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2401 ggccttttta cggttcctgg ccttttgctg gccttttgct ggccttttgc tcacatgttc
     2461 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     2521 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     2581 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     2641 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     2701 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     2761 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     2821 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     2881 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     2941 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     3001 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     3061 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagccca ccatgggagt
     3121 gcaggtggaa accatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg
     3181 cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa gttgattcct cccgggacag
     3241 aaacaagccc tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg
     3301 ggttgcccag atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta
     3361 tggtgccact gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga
     3421 gcttctaaaa ctggaaggtg ctacgcgtct gcccggttct atggtgagca agggcgagga
     3481 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     3541 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     3601 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     3661 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     3721 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     3781 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     3841 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     3901 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     3961 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     4021 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     4081 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     4141 caccggcggc atggacgagc tgtacaagcg ggctaaagtg accatgaatg acttcgacta
     4201 tctcaaactc cttggcaagg gaacctttgg caaagtcatc ctggtgcggg agaaggccac
     4261 tggccgctac tacgccatga agatcctgcg gaaggaagtc atcattgcca aggatgaagt
     4321 cgctcacaca gtcaccgaga gccgggtcct ccagaacacc aggcacccgt tcctcactgc
     4381 gctgaagtat gccttccaga cccacgaccg cctgtgcttt gtgatggagt atgccaacgg
     4441 gggtgagctg ttcttccacc tgtcccggga gcgtgtcttc acagaggagc gggcccggtt
     4501 ttatggtgca gagattgtct cggctcttga gtacttgcac tcgcgggacg tggtataccg
     4561 cgacatcaag ctggaaaacc tcatgctgga caaagatggc cacatcaaga tcactgactt
     4621 tggcctctgc aaagagggca tcagtgacgg ggccaccatg aaaaccttct gtgggacccc
     4681 ggagtacctg gcgcctgagg tgctggagga caatgactat ggccgggccg tggactggtg
     4741 ggggctgggt gtggtcatgt acgagatgat gtgcggccgc ctgcccttct acaaccagga
     4801 ccacgagcgc ctcttcgagc tcatcctcat ggaagagatc cgcttcccgc gcacgctcag
     4861 ccccgaggcc aagtccctgc ttgctgggct gcttaagaag gaccccaagc agaggcttgg
     4921 tggggggccc agcgatgcca aggaggtcat ggagcacagg ttcttcctca gcatcaactg
     4981 gcaggacgtg gtccagaaga agctcctgcc acccttcaaa cctcaggtca cgtccgaggt
     5041 cgacacaagg tacttcgatg atgaatttac cgcccagtcc atcacaatca caccccctga
     5101 ccgctatgac agcctgggct tactggagct ggaccagcgg acccacttcc cccagttctc
     5161 ctactcggcc agcatccgcg agtgaggatc caccggatct agataactga tcataatcag
     5221 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     5281 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5341 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5401 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     5461 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     5521 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     5581 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     5641 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     5701 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5761 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5821 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5881 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5941 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6001 tgcttcaata atattgaaaa aggaaga
//
