LOCUS       sequence-173573-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-tagged human Akt2(WT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  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 6117)
  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-Akt2(WT)
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        778..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1082..1285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1236..1256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1324..1333
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1330..2037
                     /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(1475..1493)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1743..1762
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3622..3743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3659..3678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3713..3732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3750..4205)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3887..3908)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4099..4118
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4232..4336
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4338..4695
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4338..4529
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4361..4381)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4546..4681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4730..5524
                     /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             4730..5524
                     /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(4784..4803)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5394..5413
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5712..5731)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5756..5803
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
      721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     1321 gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1381 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1441 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     1501 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1561 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     1621 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     1681 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     1741 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     1801 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     1861 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     1921 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     1981 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtcc
     2041 ggactcagat ctcgagctat gaatgaggtg tctgtcatca aagaaggctg gctccacaag
     2101 cgtggtgaat acatcaagac ctggaggcca cggtacttcc tgctgaagag cgacggctcc
     2161 ttcattgggt acaaggagag gcccgaggcc cctgatcaga ctctaccccc cttaaacaac
     2221 ttctccgtag cagaatgcca gctgatgaag accgagaggc cgcgacccaa cacctttgtc
     2281 atacgctgcc tgcagtggac cacagtcatc gagaggacct tccacgtgga ttctccagac
     2341 gagagggagg agtggatgcg ggccatccag atggtcgcca acagcctcaa gcagcgggcc
     2401 ccaggcgagg accccatgga ctacaagtgt ggctccccca gtgactcctc cacgactgag
     2461 gagatggaag tggcggtcag caaggcacgg gctaaagtga ccatgaatga cttcgactat
     2521 ctcaaactcc ttggcaaggg aacctttggc aaagtcatcc tggtgcggga gaaggccact
     2581 ggccgctact acgccatgaa gatcctgcgg aaggaagtca tcattgccaa ggatgaagtc
     2641 gctcacacag tcaccgagag ccgggtcctc cagaacacca ggcacccgtt cctcactgcg
     2701 ctgaagtatg ccttccagac ccacgaccgc ctgtgctttg tgatggagta tgccaacggg
     2761 ggtgagctgt tcttccacct gtcccgggag cgtgtcttca cagaggagcg ggcccggttt
     2821 tatggtgcag agattgtctc ggctcttgag tacttgcact cgcgggacgt ggtataccgc
     2881 gacatcaagc tggaaaacct catgctggac aaagatggcc acatcaagat cactgacttt
     2941 ggcctctgca aagagggcat cagtgacggg gccaccatga aaaccttctg tgggaccccg
     3001 gagtacctgg cgcctgaggt gctggaggac aatgactatg gccgggccgt ggactggtgg
     3061 gggctgggtg tggtcatgta cgagatgatg tgcggccgcc tgcccttcta caaccaggac
     3121 cacgagcgcc tcttcgagct catcctcatg gaagagatcc gcttcccgcg cacgctcagc
     3181 cccgaggcca agtccctgct tgctgggctg cttaagaagg accccaagca gaggcttggt
     3241 ggggggccca gcgatgccaa ggaggtcatg gagcacaggt tcttcctcag catcaactgg
     3301 caggacgtgg tccagaagaa gctcctgcca cccttcaaac ctcaggtcac gtccgaggtc
     3361 gacacaaggt acttcgatga tgaatttacc gcccagtcca tcacaatcac accccctgac
     3421 cgctatgaca gcctgggctt actggagctg gaccagcgga cccacttccc ccagttctcc
     3481 tactcggcca gcatccgcga gtgatctaga taactgatca taatcagcca taccacattt
     3541 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3601 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3661 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3721 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     3781 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3841 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3901 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3961 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4021 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4081 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4141 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4201 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4261 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4321 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4381 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4441 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4501 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4561 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4621 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4681 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4741 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4801 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4861 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4921 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4981 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5041 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5101 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5161 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5221 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5281 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5341 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5401 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5461 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5521 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5581 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5641 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5701 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5761 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5821 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5881 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5941 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6001 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6061 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaag
//
