LOCUS       sequence-173550-        8405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cleavable human full-length Akt1(WT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8405)
  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 8405)
  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: pFBD-Akt13C(WT)
FEATURES             Location/Qualifiers
     source          1..8405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      49..637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     538..557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     791..808
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  942..1166
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1233..1766)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1955..1960,1961..1977,1978..1983))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    2491..2539
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2564..2583
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4328..4437)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4461..4552
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4477..4494
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4601..4630
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4637..4660
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4670..4690
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             5081..5104
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    6326..6460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6363..6382)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6417..6436
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6489..6654
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6838..7292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6925..6944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7134..7155
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7318..7422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7423..7491,7492..8283)
                     /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     complement(7641..7660)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
       61 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      661 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      721 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      781 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
      841 ccttacgcat ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc
      901 ggttacggtt gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt
      961 cttaaactga acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag
     1021 ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc
     1081 taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg
     1141 ccaagggcat ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg
     1201 gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca
     1261 aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca
     1321 ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg
     1381 agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca
     1441 tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg
     1501 ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag
     1561 ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca
     1621 cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat
     1681 gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc
     1741 gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     1801 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata
     1861 acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga
     1921 ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt
     1981 caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag
     2041 cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac
     2101 gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct
     2161 gcccttgctt caggagatcg gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac
     2221 cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca
     2281 ggactctagc tatagttcta gtggttggct acgtacccgt agtggctatg gcagggcttg
     2341 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggt tggggtgggg
     2401 aaaaggaaga aacgcgggcg tattggtccc aatggggtct cggtggggta tcgacagagt
     2461 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta
     2521 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     2581 tcagttagcc tcccccatct cccggtacct cactgcacag cggcgtccgg gtggctctgg
     2641 tatcgctgcc tccaaacctc ctggatcttc ctgcaccact tgtgtgcgtt cccgctgggg
     2701 tccatcagat aatacgtcct gttaggcgtg tggacaaaga aagttttaaa attcttggcc
     2761 tctggtcgaa gttcttgtga ccaaggaatt tcacctttca gaactttgtt gacaggatcc
     2821 acataatata aatgtggtcc ttctgtgagc aacagctgtc gtcgtcttgc aaataaaccc
     2881 ttccgcttat ccactgggcc catctttagt attaaattat tttctacaaa ctggtgccaa
     2941 gggtttccgc cagcctgctt ctccaacaac aacctcttct catcttcgga aaactgtaag
     3001 tccagttcaa aggagttcga gtccagatcg tgaatgtact gctctatgtt gctgcctgac
     3061 ctctggggca ggcccgtgtc ggaggctgac agggagtgtg aggaggagga cgaagacacc
     3121 tgcatgcagc caaactggct caggagattg tcataattgc catagcagtc ctcgtcgtct
     3181 tccgacatag ccggcaggta agcggtgagc ttcggaggcg tctgctggtg caggttctcc
     3241 cacgtgacgg actcgaagaa cgggtgtgct ttaagaggtc cgtatccttc catttcctca
     3301 cagcctaacc gctttgtggc atctaaaacc aaaagtttct ccacgaggtc tcttgcctta
     3361 gggaagaatt tttctggaaa gtcatattcc aacttaatga tcttctgaaa tataagatac
     3421 tcgtttccag ctcggaatgg tgggagtcct gccacaagct ggtatattat gcatccaaga
     3481 gcccaaaggt ctgaactctt acaggcggac ttctccgtga gcagctctgg agaaacgtac
     3541 tgcgctgttc ccacgaatga gttggccctg gcttgtttgc tctctgggga taagactttt
     3601 gctgttccaa aatctgtgat ctggatgtgc atatcttcat ttaacaaaat gttttccggt
     3661 ttaaggtccc tgtgaatgat gcccttgccg tgcaagtact ctaaagcaga cacaatctca
     3721 gccgtgtaaa atcgggtaca ggtctcatcg aatgaaccga ttttgcgaat atatttaagt
     3781 agttctccat ttttggcata actaaggccg aaatacagct tctcgtcgtc ctgaaatgtg
     3841 aagtaaagct taacaaagaa ggggtgatcc aggcgcgaca tgacatcccg ctctctggtt
     3901 acatagggga ccttgttctc ttttatgata tgtcgcttct ccagaatttt aatcgcatat
     3961 tctctggagg ttgccagttc tcgagccagg acaaccgtgg aaaaagagcc ttccccaagg
     4021 attttcccaa acttgaagtc ctcaggccgc ttcttccgag gctgcggcgg aggctgggca
     4081 tgctgcaggg agccggcgcc gggccgaggc tcggctgcag tgccgtccat ggcggggccc
     4141 tgcctgctgc caccaggaat gccagggggc gtgctggact cagtctgggt cctcaccatt
     4201 gatggggatg ggcaggaaca taacaccacg ctggactgga tgggcacggc gtcatacagc
     4261 tggctggtgg tcctggccat gctagcacca tggctcgaga tcccgggtga tcaagtcttc
     4321 gtcgagtgat tgtaaataaa atgtaattta cagtatagta ttttaattaa tatacaaatg
     4381 atttgataat aattcttatt taactataat atattgtgtt gggttgaatt aaaggtccgt
     4441 atactccgga atattaatag atcatggaga taattaaaat gataaccatc tcgcaaataa
     4501 ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa atattccgga
     4561 ttattcatac cgtcccacca tcgggcgcgg atccatgtcg caccatcacc atcaccatca
     4621 ccatcaccat gcaagctgga gccacccgca gttcgaaaag ggctccggtg aaaacctgta
     4681 tttccaggcg cgcatgagcg acgtggctat tgtgaaggag ggttggctgc acaaacgagg
     4741 ggagtacatc aagacctggc ggccacgcta cttcctcctc aagaatgatg gcaccttcat
     4801 tggctacaag gagaggccgc aggatgtgga ccaacgtgag gctcccctca acaacttctc
     4861 tgtggcgcag tgccagctga tgaagacgga gcggccccgg cccaacacct tcatcatccg
     4921 ctgcctgcag tggaccactg tcatcgaacg caccttccat gtggagactc ctgaggagcg
     4981 ggaggagtgg acaaccgcca tccagactgt ggctgacggc ctcaagaagc aggaggagga
     5041 ggagatggac ttccggtcgg gctcacccag tgacaactca ctagaagttc tattccaagg
     5101 acctggggct gaagagatgg aggtgtccct ggccaagccc aagcaccgcg tgaccatgaa
     5161 cgagtttgag tacctgaagc tgctgggcaa gggcactttc ggcaaggtga tcctggtgaa
     5221 ggagaaggcc acaggccgct actacgccat gaagatcctc aagaaggaag tcatcgtggc
     5281 caaggacgag gtggcccaca cactcaccga gaaccgcgtc ctgcagaact ccaggcaccc
     5341 cttcctcaca gccctgaagt actctttcca gacccacgac cgcctctgct ttgtcatgga
     5401 gtacgccaac gggggcgagc tgttcttcca cctgtcccgg gagcgtgtgt tctccgagga
     5461 ccgggcccgc ttctatggcg ctgagattgt gtcagccctg gactacctgc actcggagaa
     5521 gaacgtggtg taccgggacc tcaagctgga gaacctcatg ctggacaagg acgggcacat
     5581 taagatcaca gacttcgggc tgtgcaagga ggggatcaag gacggtgcca ccatgaagac
     5641 cttttgcggc acacctgagt acctggcccc cgaggtgctg gaggacaatg actacggccg
     5701 tgcagtggac tggtgggggc tgggcgtggt catgtacgag atgatgtgcg gtcgcctgcc
     5761 cttctacaac caggaccatg agaagctttt tgagctcatc ctcatggagg agatccgctt
     5821 cccgcgcacg cttggtcccg aggccaagtc cttgctttca gggctgctca agaaggaccc
     5881 caagcagagg cttggcgggg gctccgagga cgccaaggag atcatgcagc atcgcttctt
     5941 tgccggtatc gtgtggcagc acgtgtacga gaagaagctc agcccaccct tcaagcccca
     6001 ggtcacgtcg gagactgaca ccaggtattt tgatgaggag ttcacggccc agatgatcac
     6061 catcacacca cctgaccaag atgacagcat ggagtgtgtg gacagcgagc gcaggcccca
     6121 cttcccccag ttctcctact cggccagcag cacggcctga gcggccgctt tcgaatctag
     6181 agcctgcagt ctcgacaagc ttgtcgagaa gtactagagg atcataatca gccataccac
     6241 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     6301 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     6361 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6421 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag
     6481 gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
     6541 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
     6601 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg
     6661 catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt
     6721 catcttcggc tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat
     6781 taatgtttgt aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg
     6841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6901 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     7021 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     7081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact
     7141 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     7201 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     7261 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     7321 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     7381 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     7441 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     7501 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     7561 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7621 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7681 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7741 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7801 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7861 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7921 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7981 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     8041 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8101 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     8161 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     8221 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     8281 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     8341 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8401 gagtt
//
