LOCUS       sequence-234746-       12794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Frb:Akt2 fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12794)
  AUTHORS   Kovalski JR, Bhaduri A, Zehnder AM, Neela PH, Che Y, Wozniak GG, 
            Khavari PA
  TITLE     The Functional Proximal Proteome of Oncogenic Ras Includes mTORC2.
  JOURNAL   Mol Cell. 2019 Jan 4. pii: S1097-2765(18)31031-1. doi: 
            10.1016/j.molcel.2018.12.001.
  PUBMED    30639242
REFERENCE   2  (bases 1 to 12794)
  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"; journalName: "Mol Cell. 
            2019 Jan 4. pii: S1097-2765(18)31031-1. doi: 
            10.1016/j.molcel.2018.12.001."
COMMENT     Sequence Label: pLEX-Frb:Akt2-IRES-Hygromycin
FEATURES             Location/Qualifiers
     source          1..12794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        2791..3094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3095..3298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3249..3269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3295..3319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3387..3665
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     misc_feature    5162..5734
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5329..5346)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5556..5575
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5737..6762
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    6769..7357
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6822..6842)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7240..7251)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7564..7797
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7898..7915)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7904..8128
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8174..8602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8261..8280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8471..8492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8611..8631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8616..8945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8796..8931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8858..8877
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8994..10019
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             8994..10019
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    10149..10270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10319..10335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10319..10335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10332..10354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10343..10359
                     /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(10367..10373,10374..10391,10392..10397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10412..10433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10550..10567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10721..11309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10801..10820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11480..12271,12272..12340))
                     /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     12103..12122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12341..12445)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12493..12627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     2761 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2821 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2881 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2941 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     3001 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     3061 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     3121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     3181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     3241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     3301 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     3361 acaccgactc tactagagga tccatgatcc tctggcatga gatgtggcat gaaggcctgg
     3421 aagaggcatc tcgtttgtac tttggggaaa ggaacgtgaa aggcatgttt gaggtgctgg
     3481 agcccttgca tgctatgatg gaacggggcc cccagactct gaaggaaaca tcctttaatc
     3541 aggcctatgg tcgagattta atggaggccc aagagtggtg caggaagtac atgaaatcag
     3601 ggaatgtcaa ggacctcctc caagcctggg acctctatta tcatgtgttc cgacgaatct
     3661 caaagacgcg tgccaccatg aatgaggtgt ctgtcatcaa agaaggctgg ctccacaagc
     3721 gtggtgaata catcaagacc tggaggccac ggtacttcct gctgaagagc gacggctcct
     3781 tcattgggta caaggagagg cccgaggccc ctgatcagac tctacccccc ttaaacaact
     3841 tctccgtagc agaatgccag ctgatgaaga ccgagaggcc gcgacccaac acctttgtca
     3901 tacgctgcct gcagtggacc acagtcatcg agaggacctt ccacgtggat tctccagacg
     3961 agagggagga gtggatgcgg gccatccaga tggtcgccaa cagcctcaag cagcgggccc
     4021 caggcgagga ccccatggac tacaagtgtg gctcccccag tgactcctcc acgactgagg
     4081 agatggaagt ggcggtcagc aaggcacggg ctaaagtgac catgaatgac ttcgactatc
     4141 tcaaactcct tggcaaggga acctttggca aagtcatcct ggtgcgggag aaggccactg
     4201 gccgctacta cgccatgaag atcctgcgga aggaagtcat cattgccaag gatgaagtcg
     4261 ctcacacagt caccgagagc cgggtcctcc agaacaccag gcacccgttc ctcactgcgc
     4321 tgaagtatgc cttccagacc cacgaccgcc tgtgctttgt gatggagtat gccaacgggg
     4381 gtgagctgtt cttccacctg tcccgggagc gtgtcttcac agaggagcgg gcccggtttt
     4441 atggtgcaga gattgtctcg gctcttgagt acttgcactc gcgggacgtg gtataccgcg
     4501 acatcaagct ggaaaacctc atgctggaca aagatggcca catcaagatc actgactttg
     4561 gcctctgcaa agagggcatc agtgacgggg ccaccatgaa aaccttctgt gggaccccgg
     4621 agtacctggc gcctgaggtg ctggaggaca atgactatgg ccgggccgtg gactggtggg
     4681 ggctgggtgt ggtcatgtac gagatgatgt gcggccgcct gcccttctac aaccaggacc
     4741 acgagcgcct cttcgagctc atcctcatgg aagagatccg cttcccgcgc acgctcagcc
     4801 ccgaggccaa gtccctgctt gctgggctgc ttaagaagga ccccaagcag aggcttggtg
     4861 gggggcccag cgatgccaag gaggtcatgg agcacaggtt cttcctcagc atcaactggc
     4921 aggacgtggt ccagaagaag ctcctgccac ccttcaaacc tcaggtcacg tccgaggtcg
     4981 acacaaggta cttcgatgat gaatttaccg cccagtccat cacaatcaca ccccctgacc
     5041 gctatgacag cctgggctta ctggagctgg accagcggac ccacttcccc cagttctcct
     5101 actcggccag catccgcgat aactcgagtc tagagggccc gacgcgtctg gccaccggtc
     5161 gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     5221 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     5281 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     5341 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     5401 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     5461 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     5521 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     5581 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     5641 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     5701 gggacgtggt tttcctttga aaaacacgat aataccatga agaaacctga actgacagca
     5761 acttctgttg agaagtttct cattgaaaaa tttgattctg tttctgatct catgcagctg
     5821 tctgaaggtg aagaaagcag agccttttct tttgatgttg gaggaagagg ttatgttctg
     5881 agggtcaatt cttgtgctga tggtttttac aaagacagat atgtttacag acactttgcc
     5941 tctgctgctc tgccaattcc agaagttctg gacattggag aattttctga atctctcacc
     6001 tactgcatca gcagaagagc acaaggagtc actctccagg atctccctga aactgagctg
     6061 ccagctgttc tgcaacctgt tgctgaagca atggatgcca ttgcagcagc tgatctgagc
     6121 caaacctctg gatttggtcc ttttggtccc caaggcattg gtcagtacac cacttggagg
     6181 gatttcattt gtgccattgc tgatcctcat gtctatcact ggcagactgt gatggatgac
     6241 acagtttctg cttctgttgc tcaggcactg gatgaactca tgctgtgggc agaagattgt
     6301 cctgaagtca gacacctggt ccatgctgat tttggaagca acaatgttct gacagacaat
     6361 ggcagaatca ctgcagtcat tgactggtct gaagccatgt ttggagattc tcaatatgag
     6421 gttgccaaca tttttttttg gagaccttgg ctggcttgca tggaacaaca aacaagatat
     6481 tttgaaagaa gacacccaga actggctggt tcccccagac tgagagccta catgctcaga
     6541 attggcctgg accaactgta tcaatctctg gttgatggaa actttgatga tgctgcttgg
     6601 gcacaaggaa gatgtgatgc cattgtgagg tctggtgctg gaactgttgg aagaactcaa
     6661 attgcaagaa ggtctgctgc tgtttggact gatggatgtg ttgaagttct ggctgactct
     6721 ggaaacagga gaccctccac aagacccaga gccaaggaat gagttaacaa tcaacctctg
     6781 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     6841 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     6901 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6961 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     7021 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     7081 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     7141 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     7201 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     7261 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     7321 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     7381 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     7441 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     7501 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg
     7561 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     7621 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7681 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7741 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     7801 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     7861 gtgagaggcc ttgacattgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     7921 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     7981 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     8041 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     8101 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     8161 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     8221 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     8281 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     8341 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     8401 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     8461 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     8521 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     8581 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     8641 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     8701 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     8761 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     8821 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     8881 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     8941 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgatgaaaa
     9001 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     9061 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     9121 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     9181 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
     9241 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     9301 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     9361 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     9421 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     9481 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     9541 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     9601 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     9661 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     9721 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     9781 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     9841 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     9901 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     9961 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc
    10021 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
    10081 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
    10141 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
    10201 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
    10261 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
    10321 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
    10381 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
    10441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
    10501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
    10561 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
    10621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
    10681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
    10741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
    10801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
    10861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
    10921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
    10981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
    11041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
    11101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
    11161 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
    11221 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
    11281 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
    11341 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
    11401 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
    11461 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
    11521 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
    11581 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
    11641 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
    11701 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
    11761 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
    11821 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
    11881 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
    11941 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
    12001 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
    12061 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
    12121 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
    12181 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
    12241 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
    12301 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
    12361 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
    12421 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
    12481 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    12541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    12601 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
    12661 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
    12721 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
    12781 acaaacttag tagt
//
