LOCUS       sequence-174182-        6477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable Akt2(K14A) mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6477)
  AUTHORS   Ebner M, Sinkovics B, Szczygiel M, Ribeiro DW, Yudushkin I
  TITLE     Localization of mTORC2 activity inside cells.
  JOURNAL   J Cell Biol. 2017 Jan 31. pii: jcb.201610060. doi: 
            10.1083/jcb.201610060.
  PUBMED    28143890
REFERENCE   2  (bases 1 to 6477)
  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"; journalName: "J Cell 
            Biol. 2017 Jan 31. pii: jcb.201610060. doi: 10.1083/jcb.201610060."
COMMENT     Sequence Label: C1-FKBP(F36V):mCherry-Akt2(K14A)
FEATURES             Location/Qualifiers
     source          1..6477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             192..986
                     /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             192..986
                     /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(246..265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     856..875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1174..1193)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1218..1265
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1594..2182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2083..2102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2368..2671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2672..2875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2826..2846
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2909..2911,2912..3232)
                     /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             3257..3964
                     /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(3402..3420)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3670..3689
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5561..5682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5598..5617)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5652..5671
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5689..6144)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5826..5847)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6038..6057
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6171..6275
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6277..6477,1..157)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6277..6468
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6300..6320)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
       61 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
      121 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
      181 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      241 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      301 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      361 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      421 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      481 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      541 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      601 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
      661 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
      721 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
      781 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
      841 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
      901 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcacagcgc atcgccttct
      961 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1021 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1081 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1141 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     1201 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     1261 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     1321 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     1381 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     1441 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1501 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1561 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1621 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1681 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1741 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1801 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1861 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1921 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1981 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2041 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2101 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2161 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2221 ttgctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2281 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     2341 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2401 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2461 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2521 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2581 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2641 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2701 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2761 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2821 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     2881 agtgaaccgt cagatccgct agcccaccat gggagtgcag gtggagacta tctccccagg
     2941 agacgggcgc accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct
     3001 tgaagatgga aagaaagttg attcctcccg ggacagaaac aagcccttta agtttatgct
     3061 aggcaagcag gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca
     3121 gagagccaaa ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat
     3181 catcccacca catgccactc tcgtcttcga tgtggagctt ctaaaactgg aaggtgctac
     3241 gcgtctgccc ggttctatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     3301 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3361 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3421 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     3481 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     3541 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3601 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3661 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     3721 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     3781 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3841 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     3901 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3961 caagtccgga ctcagatctc gagctatgaa tgaggtgtct gtcatcaaag aaggctggct
     4021 ccacgcacgt ggtgaataca tcaagacctg gaggccacgg tacttcctgc tgaagagcga
     4081 cggctccttc attgggtaca aggagaggcc cgaggcccct gatcagactc tacccccctt
     4141 aaacaacttc tccgtagcag aatgccagct gatgaagacc gagaggccgc gacccaacac
     4201 ctttgtcata cgctgcctgc agtggaccac agtcatcgag aggaccttcc acgtggattc
     4261 tccagacgag agggaggagt ggatgcgggc catccagatg gtcgccaaca gcctcaagca
     4321 gcgggcccca ggcgaggacc ccatggacta caagtgtggc tcccccagtg actcctccac
     4381 gactgaggag atggaagtgg cggtcagcaa ggcacgggct aaagtgacca tgaatgactt
     4441 cgactatctc aaactccttg gcaagggaac ctttggcaaa gtcatcctgg tgcgggagaa
     4501 ggccactggc cgctactacg ccatgaagat cctgcggaag gaagtcatca ttgccaagga
     4561 tgaagtcgct cacacagtca ccgagagccg ggtcctccag aacaccaggc acccgttcct
     4621 cactgcgctg aagtatgcct tccagaccca cgaccgcctg tgctttgtga tggagtatgc
     4681 caacgggggt gagctgttct tccacctgtc ccgggagcgt gtcttcacag aggagcgggc
     4741 ccggttttat ggtgcagaga ttgtctcggc tcttgagtac ttgcactcgc gggacgtggt
     4801 ataccgcgac atcaagctgg aaaacctcat gctggacaaa gatggccaca tcaagatcac
     4861 tgactttggc ctctgcaaag agggcatcag tgacggggcc accatgaaaa ccttctgtgg
     4921 gaccccggag tacctggcgc ctgaggtgct ggaggacaat gactatggcc gggccgtgga
     4981 ctggtggggg ctgggtgtgg tcatgtacga gatgatgtgc ggccgcctgc ccttctacaa
     5041 ccaggaccac gagcgcctct tcgagctcat cctcatggaa gagatccgct tcccgcgcac
     5101 gctcagcccc gaggccaagt ccctgcttgc tgggctgctt aagaaggacc ccaagcagag
     5161 gcttggtggg gggcccagcg atgccaagga ggtcatggag cacaggttct tcctcagcat
     5221 caactggcag gacgtggtcc agaagaagct cctgccaccc ttcaaacctc aggtcacgtc
     5281 cgaggtcgac acaaggtact tcgatgatga atttaccgcc cagtccatca caatcacacc
     5341 ccctgaccgc tatgacagcc tgggcttact ggagctggac cagcggaccc acttccccca
     5401 gttctcctac tcggccagca tccgcgagtg aggatccacc ggatctagat aactgatcat
     5461 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5521 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5581 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5641 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     5701 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5761 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5821 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5881 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5941 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6001 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6061 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6121 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6181 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6241 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     6301 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6361 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     6421 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagccccgc ccctaac
//
