LOCUS       sequence-134142-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human AKT3 K177M.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  AUTHORS   Baek ST, Copeland B, Yun EJ, Kwon SK, Guemez-Gamboa A, Schaffer AE, 
            Kim S, Kang HC, Song S, Mathern GW, Gleeson JG
  TITLE     An AKT3-FOXG1-reelin network underlies defective migration in human 
            focal malformations of cortical development.
  JOURNAL   Nat Med. 2015 Dec;21(12):1445-54. doi: 10.1038/nm.3982. Epub 2015 
            Nov 2.
  PUBMED    26523971
REFERENCE   2  (bases 1 to 7647)
  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: "10.1038/nm.3982"; 
            journalName: "Nat Med"; date: "2015-12"; volume: "21"; issue: "12"; 
            pages: "1445-54"
COMMENT     Sequence Label: pCIG AKT3 K177M
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(122..141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(801..1592,1593..1661))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1424..1443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1662..1766)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1797..2176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2178..2455
                     /label=chicken beta-actin promoter
     intron          2456..3468
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3392..3414
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3476..3495
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(5033..5607,5608..5610,5611..5619)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5200..5217)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5427..5446
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5620..5622,5623..5625,5626..6339)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5620..5622,5623..5625,5626..6336)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5665..5686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5926..5945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6273..6294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6431..6450)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6496..6551
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6550..6569)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6942..6958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6942..6958)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6955..6977)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6966..6982
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6990..6996,6997..7014,7015..7020))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7035..7056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7115..7310
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7161..7296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7316..7450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7353..7372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7407..7426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7518..7535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agnacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2161 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     2221 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     2281 ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
     2341 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     2401 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     2461 cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct
     2521 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     2581 taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa
     2641 agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg
     2701 tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg
     2761 cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc
     2821 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     2881 ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc
     2941 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     3001 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     3061 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     3121 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     3181 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     3241 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     3301 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     3361 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     3421 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     3481 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctcgagct caagcttcga
     3541 attcatgagc gatgttacca ttgtgaaaga aggttgggtt cagaagaggg gagaatatat
     3601 aaaaaactgg aggccaagat acttcctttt gaagacagat ggctcattca taggatataa
     3661 agagaaacct caagatgtgg atttacctta tcccctcaac aacttttcag tggcaaaatg
     3721 ccagttaatg aaaacagaac gaccaaagcc aaacacattt ataatcagat gtctccagtg
     3781 gactactgtt atagagagaa catttcatgt agatactcca gaggaaaggg aagaatggac
     3841 agaagctatc caggctgtag cagacagact gcagaggcaa gaagaggaga gaatgaattg
     3901 tagtccaact tcacaaattg ataatatagg agaggaagag atggatgcct ctacaaccca
     3961 tcataaaaga aagacaatga atgattttga ctatttgaaa ctactaggta aaggcacttt
     4021 tgggaaagtt attttggttc gagagaaggc aagtggaaaa tactatgcta tgaagattct
     4081 gaagaaagaa gtcattattg caaaggatga agtggcacac actctaactg aaagcagagt
     4141 attaaagaac actagacatc cctttttaac atccttgaaa tattccttcc agacaaaaga
     4201 ccgtttgtgt tttgtgatgg aatatgttaa tgggggcgag ctgtttttcc atttgtcgag
     4261 agagcgggtg ttctctgagg accgcacacg tttctatggt gcagaaattg tctctgcctt
     4321 ggactatcta cattccggaa agattgtgta ccgtgatctc aagttggaga atctaatgct
     4381 ggacaaagat ggccacataa aaattacaga ttttggactt tgcaaagaag ggatcacaga
     4441 tgcagccacc atgaagacat tctgtggcac tccagaatat ctggcaccag aggtgttaga
     4501 agataatgac tatggccgag cagtagactg gtggggccta ggggttgtca tgtatgaaat
     4561 gatgtgtggg aggttacctt tctacaacca ggaccatgag aaactttttg aattaatatt
     4621 aatggaagac attaaatttc ctcgaacact ctcttcagat gcaaaatcat tgctttcagg
     4681 gctcttgata aaggatccaa ataaacgcct tggtggagga ccagatgatg caaaagaaat
     4741 tatgagacac agtttcttct ctggagtaaa ctggcaagat gtatatgata aaaagcttgt
     4801 acctcctttt aaacctcaag taacatctga gacagatact agatattttg atgaagaatt
     4861 tacagctcag actattacaa taacaccacc tgaaaaatat gatgaggatg gtatggactg
     4921 catggacaat gagaggcggc cgcatttccc tcaattttcc tactctgcaa gtggacgaga
     4981 ataagaataa gggcgaattc tgcagtcgac ggtaccgcgg gcccgggatc cgcccctctc
     5041 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     5101 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     5161 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     5221 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     5281 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     5341 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     5401 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     5461 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     5521 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     5581 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     5641 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     5701 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     5761 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     5821 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     5881 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     5941 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     6001 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     6061 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     6121 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     6181 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     6241 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     6301 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcaa ttcactcctc
     6361 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     6421 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     6481 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     6541 tgtgtctatc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     6601 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     6661 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     6721 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     6781 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     6841 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcac cgtcgaccag
     6901 ctggtcgacg gtgcaccgtc gaccagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6961 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7021 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7081 ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag
     7141 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     7201 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     7261 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt
     7321 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7381 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7441 tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7621 ggaaagaaca tgtgagcaaa aggccag
//
