LOCUS       sequence-228989-       12249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-birA*-N-Ras Q61K/Y40A fusion protein in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12249)
  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 12249)
  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-uORF-HA-birA*-N-Ras(Q61K/Y40A)-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..12249
                     /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             3411..3437
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(3438..3785,3786..3788,3789..4397)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     misc_feature    5040..5612
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5207..5224)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5434..5453
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5615..6217
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6114..6134
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6224..6812
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6277..6297)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6695..6706)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7019..7252
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7353..7370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7359..7583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7629..8057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7716..7735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7926..7947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8066..8086)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8071..8400
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8251..8386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8313..8332
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8449..9474
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             8449..9474
                     /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    9604..9725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9774..9790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9774..9790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9787..9809)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9798..9814
                     /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(9822..9828,9829..9846,9847..9852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9867..9888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10005..10022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10176..10764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10256..10275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10935..11726,11727..11795))
                     /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     11558..11577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11796..11900)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    11948..12082
                     /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 tctaccatgg cataacgact aatatttatg tacccatacg
     3421 atgtgcctga ctatgctaag gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca
     3481 acggcgagtt ccactctggc gagcagctgg gagagaccct gggaatgagc agagccgcca
     3541 tcaacaagca catccagaca ctgagagact ggggagtgga cgtgttcacc gtgcctggca
     3601 agggctacag cctgcctgag cctatccagc tgctgaacgc caagcagatc ctgggacagc
     3661 tggatggcgg aagcgtggcc gtgctgcctg tgatcgactc caccaatcag tacctgctgg
     3721 acagaatcgg agagctgaag tccggcgacg cctgcatcgc cgagtaccag caggctggca
     3781 gaggaggcag aggacggaag tggttcagcc cattcggagc caacctgtac ctgtccatgt
     3841 tctggagact ggagcaggga cctgctgctg ccatcggact gagtctggtg atcggaatcg
     3901 tgatggccga ggtgctgaga aagctgggag ccgacaaggt gagagtgaag tggcctaatg
     3961 acctgtacct ccaggaccgc aagctggctg gcatcctggt ggagctgaca ggcaagacag
     4021 gcgatgccgc tcagatcgtg atcggagccg gaatcaacat ggccatgaga agagtggagg
     4081 agagcgtggt gaaccagggc tggatcaccc tgcaggaggc tggcatcaac ctggaccgga
     4141 acaccctggc cgccatgctg atcagagagc tgagagccgc tctggagctg ttcgagcagg
     4201 agggactggc tccttacctg agcagatggg agaagctgga caacttcatc aacagacctg
     4261 tgaagctgat catcggcgac aaggaaatct tcggcatctc cagaggaatc gacaagcagg
     4321 gagctctgct gctggagcag gacggaatca tcaagccctg gatgggcgga gaaatctccc
     4381 tgagaagcgc agagaaggga tccagcggcg gcggcagcgg cggcggcggc agcactgagt
     4441 acaaactggt ggtggttgga gcaggtggtg ttgggaaaag cgcactgaca atccagctaa
     4501 tccagaacca ctttgtagat gaatatgatc ccaccataga ggattctgcc agaaaacaag
     4561 tggttataga tggtgaaacc tgtttgttgg acatactgga tacagctgga aaagaagagt
     4621 acagtgccat gagagaccaa tacatgagga caggcgaagg cttcctctgt gtatttgcca
     4681 tcaataatag caagtcattt gcggatatta acctctacag ggagcagatt aagcgagtaa
     4741 aagactcgga tgatgtacct atggtgctag tgggaaacaa gtgtgatttg ccaacaagga
     4801 cagttgatac aaaacaagcc cacgaactgg ccaagagtta cgggattcca ttcattgaaa
     4861 cctcagccaa gaccagacag ggtgttgaag atgcttttta cacactggta agagaaatac
     4921 gccagtaccg aatgaaaaaa ctcaacagca gtgatgatgg gactcagggt tgtatgggat
     4981 tgccatgtgt ggtgatgtaa ctcgagtcta gagggcccga cgcgtctggc caccggtcgc
     5041 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     5101 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     5161 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     5221 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     5281 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     5341 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     5401 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     5461 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     5521 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     5581 gacgtggttt tcctttgaaa aacacgataa taccatggcc accgagtaca agcccacggt
     5641 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     5701 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     5761 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     5821 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     5881 cgccgagatc ggctcgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5941 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     6001 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     6061 ggaggcggcc gagcgcgctg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     6121 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     6181 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagtt aacaatcaac ctctggatta
     6241 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     6301 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     6361 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6421 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6481 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6541 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6601 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     6661 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6721 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6781 gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta
     6841 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     6901 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     6961 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg
     7021 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     7081 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     7141 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7201 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     7261 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     7321 aggccttgac attgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     7381 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     7441 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     7501 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     7561 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     7621 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     7681 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     7741 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     7801 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     7861 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     7921 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     7981 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     8041 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     8101 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     8161 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     8221 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     8281 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     8341 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     8401 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgat gaaaaagcct
     8461 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     8521 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     8581 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     8641 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc
     8701 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     8761 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg
     8821 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     8881 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     8941 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     9001 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     9061 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     9121 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     9181 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     9241 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     9301 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     9361 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     9421 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgtg
     9481 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     9541 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     9601 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     9661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     9721 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     9781 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     9841 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     9901 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     9961 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
    10021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
    10081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
    10141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    10201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    10261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    10321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    10381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
    10441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    10501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    10561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    10621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    10681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    10741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    10801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
    10861 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
    10921 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
    10981 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
    11041 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
    11101 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
    11161 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
    11221 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
    11281 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
    11341 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
    11401 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
    11461 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
    11521 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
    11581 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
    11641 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    11701 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    11761 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    11821 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    11881 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
    11941 ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    12001 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    12061 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
    12121 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
    12181 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
    12241 cttagtagt
//
