LOCUS       sequence-173590-        6042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-tagged human Akt1(WT).
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-Akt1(WT)
FEATURES             Location/Qualifiers
     source          1..6042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      26..614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     515..534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        789..1092
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1093..1296
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1247..1267
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1335..1344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1341..2048
                     /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(1486..1504)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1754..1773
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2070..3509
                     /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    3630..3751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3667..3686)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3721..3740
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3758..4213)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3895..3916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4107..4126
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4240..4344
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4369..4389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4413..4623
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4435..4631
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4482..4617
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4666..5460
                     /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             4666..5460
                     /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(4720..4739)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5330..5349
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5648..5667)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5692..5739
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
       61 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      121 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      181 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      241 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      301 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      361 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      421 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      481 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      541 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      601 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      661 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      721 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      781 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      841 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      901 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      961 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1021 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     1081 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     1141 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1201 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     1261 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
     1321 tagcgctacc ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca
     1381 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     1441 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1501 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     1561 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     1621 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     1681 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     1741 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     1801 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     1861 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     1921 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     1981 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2041 tgtacaagtc cggactcaga tctcgagcta tgagcgacgt ggctattgtg aaggagggtt
     2101 ggctgcacaa acgaggggag tacatcaaga cctggcggcc acgctacttc ctcctcaaga
     2161 atgatggcac cttcattggc tacaaggagc ggccgcagga tgtggaccaa cgtgaggctc
     2221 ccctcaacaa cttctctgtg gcgcagtgcc agctgatgaa gacggagcgg ccccggccca
     2281 acaccttcat catccgctgc ctgcagtgga ccactgtcat cgaacgcacc ttccatgtgg
     2341 agactcctga ggagcgggag gagtggacaa ccgccatcca gactgtggct gacggcctca
     2401 agaagcagga ggaggaggag atggacttcc ggtcgggctc acccagtgac aactcagggg
     2461 ctgaagagat ggaggtgtcc ctggccaagc ccaagcaccg cgtgaccatg aacgagtttg
     2521 agtacctgaa gctgctgggc aagggcactt tcggcaaggt gatcctggtg aaggagaagg
     2581 ccacaggccg ctactacgcc atgaagatcc tcaagaagga agtcatcgtg gccaaggacg
     2641 aggtggccca cacactcacc gagaaccgcg tcctgcagaa ctccaggcac cccttcctca
     2701 cagccctgaa gtactctttc cagacccacg accgcctctg ctttgtcatg gagtacgcca
     2761 acgggggcga gctgttcttc cacctgtccc gggagcgtgt gttctccgag gaccgggccc
     2821 gcttctatgg cgctgagatt gtgtcagccc tggactacct gcactcggag aagaacgtgg
     2881 tgtaccggga cctcaagctg gagaacctca tgctggacaa ggacgggcac attaagatca
     2941 cagacttcgg gctgtgcaag gaggggatca aggacggtgc caccatgaag accttttgcg
     3001 gcacacctga gtacctggcc cccgaggtgc tggaggacaa tgactacggc cgtgcagtgg
     3061 actggtgggg gctgggcgtg gtcatgtacg agatgatgtg cggtcgcctg cccttctaca
     3121 accaggacca tgagaagctt tttgagctca tcctcatgga ggagatccgc ttcccgcgca
     3181 cgcttggtcc cgaggccaag tccttgcttt cagggctgct caagaaggac cccaagcaga
     3241 ggcttggcgg gggctccgag gacgccaagg agatcatgca gcatcgcttc tttgccggta
     3301 tcgtgtggca gcacgtgtac gagaagaagc tcagcccacc cttcaagccc caggtcacgt
     3361 cggagactga caccaggtat tttgatgagg agttcacggc ccagatgatc accatcacac
     3421 cacctgacca agatgacagc atggagtgtg tggacagcga gcgcaggccc cacttccccc
     3481 agttctccta ctcggccagc ggcacggcct gatctagata actgatcata atcagccata
     3541 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3601 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3661 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3721 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     3781 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3841 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3901 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3961 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4021 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4081 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4141 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4201 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4261 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4321 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4381 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4501 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4561 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4621 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4681 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4741 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4801 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4861 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4921 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4981 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5041 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5101 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5161 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5221 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5281 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5341 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5401 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5461 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5521 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5581 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5641 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5701 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5761 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5821 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5881 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5941 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6001 aaatccctta acgtgagttt tcgttccact gagcgtcaga cc
//
