LOCUS       Exported                8381 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses human full-length Akt1(WT).
ACCESSION   .
VERSION     .
KEYWORDS    pFBD-Akt1(WT)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8381)
  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 8381)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8381
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     753..770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  904..1128
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1195..1728)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1917..1945)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    2453..2501
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2526..2545
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4290..4399)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4423..4514
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4439..4456
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4563..4592
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4599..4622
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4632..4652
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQA"
     CDS             4656..6095
                     /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
                     ECVDSERRPHFPQFSYSASSTA"
     polyA_signal    6264..6398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6301..6320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6355..6374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6427..6592
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6776..7230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6863..6882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7072..7093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7256..7360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7361..8221
                     /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(7579..7598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      721 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      781 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagacc
      841 agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc
      901 gggtgtgggc ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa
      961 agtcttaaac tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag
     1021 catactggac ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc
     1081 gtcgtattaa agaggggcgt ggccaagggc atggtaaaga ctatattcgc ggcgttgtga
     1141 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     1201 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
     1261 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
     1321 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
     1381 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacttggg
     1441 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
     1501 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
     1561 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
     1621 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
     1681 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
     1741 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     1801 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     1861 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
     1921 tacgaaccga acaggcttat gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc
     1981 gtcacccggc aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg
     2041 agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc ggccttgctg ttcttctacg
     2101 gcaaggtgct gtgcacggat ctgcccttgc ttcaggagat cggtagacct cggccgtcgc
     2161 ggcgcttgcc ggtggtgctg accccggatg aagtggttcg catcctcggt tttctggaag
     2221 gcgagcatcg tttgttcgcc caggactcta gctatagttc tagtggttgg ctacgtaccc
     2281 gtagtggcta tggcagggct tgccgccccg acgttggctg cgagccctgg gccttcaccc
     2341 gaacttgggg gttggggtgg ggaaaaggaa gaaacgcggg cgtattggtc ccaatggggt
     2401 ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac
     2461 gacccaacac ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc
     2521 cggtattgtc tccttccgtg tttcagttag cctcccccat ctcccggtac ctcactgcac
     2581 agcggcgtcc gggtggctct ggtatcgctg cctccaaacc tcctggatct tcctgcacca
     2641 cttgtgtgcg ttcccgctgg ggtccatcag ataatacgtc ctgttaggcg tgtggacaaa
     2701 gaaagtttta aaattcttgg cctctggtcg aagttcttgt gaccaaggaa tttcaccttt
     2761 cagaactttg ttgacaggat ccacataata taaatgtggt ccttctgtga gcaacagctg
     2821 tcgtcgtctt gcaaataaac ccttccgctt atccactggg cccatcttta gtattaaatt
     2881 attttctaca aactggtgcc aagggtttcc gccagcctgc ttctccaaca acaacctctt
     2941 ctcatcttcg gaaaactgta agtccagttc aaaggagttc gagtccagat cgtgaatgta
     3001 ctgctctatg ttgctgcctg acctctgggg caggcccgtg tcggaggctg acagggagtg
     3061 tgaggaggag gacgaagaca cctgcatgca gccaaactgg ctcaggagat tgtcataatt
     3121 gccatagcag tcctcgtcgt cttccgacat agccggcagg taagcggtga gcttcggagg
     3181 cgtctgctgg tgcaggttct cccacgtgac ggactcgaag aacgggtgtg ctttaagagg
     3241 tccgtatcct tccatttcct cacagcctaa ccgctttgtg gcatctaaaa ccaaaagttt
     3301 ctccacgagg tctcttgcct tagggaagaa tttttctgga aagtcatatt ccaacttaat
     3361 gatcttctga aatataagat actcgtttcc agctcggaat ggtgggagtc ctgccacaag
     3421 ctggtatatt atgcatccaa gagcccaaag gtctgaactc ttacaggcgg acttctccgt
     3481 gagcagctct ggagaaacgt actgcgctgt tcccacgaat gagttggccc tggcttgttt
     3541 gctctctggg gataagactt ttgctgttcc aaaatctgtg atctggatgt gcatatcttc
     3601 atttaacaaa atgttttccg gtttaaggtc cctgtgaatg atgcccttgc cgtgcaagta
     3661 ctctaaagca gacacaatct cagccgtgta aaatcgggta caggtctcat cgaatgaacc
     3721 gattttgcga atatatttaa gtagttctcc atttttggca taactaaggc cgaaatacag
     3781 cttctcgtcg tcctgaaatg tgaagtaaag cttaacaaag aaggggtgat ccaggcgcga
     3841 catgacatcc cgctctctgg ttacataggg gaccttgttc tcttttatga tatgtcgctt
     3901 ctccagaatt ttaatcgcat attctctgga ggttgccagt tctcgagcca ggacaaccgt
     3961 ggaaaaagag ccttccccaa ggattttccc aaacttgaag tcctcaggcc gcttcttccg
     4021 aggctgcggc ggaggctggg catgctgcag ggagccggcg ccgggccgag gctcggctgc
     4081 agtgccgtcc atggcggggc cctgcctgct gccaccagga atgccagggg gcgtgctgga
     4141 ctcagtctgg gtcctcacca ttgatgggga tgggcaggaa cataacacca cgctggactg
     4201 gatgggcacg gcgtcataca gctggctggt ggtcctggcc atgctagcac catggctcga
     4261 gatcccgggt gatcaagtct tcgtcgagtg attgtaaata aaatgtaatt tacagtatag
     4321 tattttaatt aatatacaaa tgatttgata ataattctta tttaactata atatattgtg
     4381 ttgggttgaa ttaaaggtcc gtatactccg gaatattaat agatcatgga gataattaaa
     4441 atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa
     4501 aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc ggatccatgt
     4561 cgcaccatca ccatcaccat caccatcacc atgcaagctg gagccacccg cagttcgaaa
     4621 agggctccgg tgaaaacctg tatttccagg cgcgcatgag cgacgtggct attgtgaagg
     4681 agggttggct gcacaaacga ggggagtaca tcaagacctg gcggccacgc tacttcctcc
     4741 tcaagaatga tggcaccttc attggctaca aggagaggcc gcaggatgtg gaccaacgtg
     4801 aggctcccct caacaacttc tctgtggcgc agtgccagct gatgaagacg gagcggcccc
     4861 ggcccaacac cttcatcatc cgctgcctgc agtggaccac tgtcatcgaa cgcaccttcc
     4921 atgtggagac tcctgaggag cgggaggagt ggacaaccgc catccagact gtggctgacg
     4981 gcctcaagaa gcaggaggag gaggagatgg acttccggtc gggctcaccc agtgacaact
     5041 caggggctga agagatggag gtgtccctgg ccaagcccaa gcaccgcgtg accatgaacg
     5101 agtttgagta cctgaagctg ctgggcaagg gcactttcgg caaggtgatc ctggtgaagg
     5161 agaaggccac aggccgctac tacgccatga agatcctcaa gaaggaagtc atcgtggcca
     5221 aggacgaggt ggcccacaca ctcaccgaga accgcgtcct gcagaactcc aggcacccct
     5281 tcctcacagc cctgaagtac tctttccaga cccacgaccg cctctgcttt gtcatggagt
     5341 acgccaacgg gggcgagctg ttcttccacc tgtcccggga gcgtgtgttc tccgaggacc
     5401 gggcccgctt ctatggcgct gagattgtgt cagccctgga ctacctgcac tcggagaaga
     5461 acgtggtgta ccgggacctc aagctggaga acctcatgct ggacaaggac gggcacatta
     5521 agatcacaga cttcgggctg tgcaaggagg ggatcaagga cggtgccacc atgaagacct
     5581 tttgcggcac acctgagtac ctggcccccg aggtgctgga ggacaatgac tacggccgtg
     5641 cagtggactg gtgggggctg ggcgtggtca tgtacgagat gatgtgcggt cgcctgccct
     5701 tctacaacca ggaccatgag aagctttttg agctcatcct catggaggag atccgcttcc
     5761 cgcgcacgct tggtcccgag gccaagtcct tgctttcagg gctgctcaag aaggacccca
     5821 agcagaggct tggcgggggc tccgaggacg ccaaggagat catgcagcat cgcttctttg
     5881 ccggtatcgt gtggcagcac gtgtacgaga agaagctcag cccacccttc aagccccagg
     5941 tcacgtcgga gactgacacc aggtattttg atgaggagtt cacggcccag atgatcacca
     6001 tcacaccacc tgaccaagat gacagcatgg agtgtgtgga cagcgagcgc aggccccact
     6061 tcccccagtt ctcctactcg gccagcagca cggcctgagc ggccgctttc gaatctagag
     6121 cctgcagtct cgacaagctt gtcgagaagt actagaggat cataatcagc cataccacat
     6181 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     6241 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     6301 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6361 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga
     6421 gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     6481 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     6541 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     6601 tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca
     6661 tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta
     6721 atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga atgggacgcg
     6781 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6841 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6901 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6961 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     7021 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct
     7081 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7141 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7201 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     7261 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7321 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7381 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7441 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7501 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7561 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7621 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7681 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7741 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7801 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7861 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7921 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7981 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8041 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8101 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8161 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8221 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8281 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     8341 gttttcgttc cactgagcgt cagaccccgt agaaaagatc a
//
