LOCUS       sequence-234082-       12225 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses HA-birA*-K-Ras G12D fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12225)
  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 12225)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-HA-birA*-K-Ras(G12D)-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..12225
                     /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             3390..3416
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(3417..3764,3765..3767,3768..4376)
                     /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"
     CDS             join(4413..4442,4443..4445,4446..4976)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="TEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVI
                     DGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVKD
                     SEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREIR
                     KHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             4413..4976
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="TEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVI
                     DGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVKD
                     SEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREIR
                     KHKEKMSKDGKKKKKKSKTKCVIM"
     misc_feature    5016..5588
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5183..5200)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5410..5429
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5591..6193
                     /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     6090..6110
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6200..6788
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6253..6273)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6671..6682)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6995..7228
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7329..7346)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7335..7559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7605..8033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7692..7711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7902..7923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8042..8062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8047..8376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8227..8362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8289..8308
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8425..9450
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             8425..9450
                     /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    9580..9701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9750..9766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9750..9766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9763..9785)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9774..9790
                     /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(9798..9804,9805..9822,9823..9828))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9843..9864
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9981..9998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10152..10740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10232..10251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10911..11702,11703..11771))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALILFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11534..11553
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11772..11876)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    11924..12058
                     /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 tctaccatgt acccatacga tgtgcctgac tatgctaagg
     3421 acaacaccgt gcccctgaag ctgatcgccc tgctggccaa cggcgagttc cactctggcg
     3481 agcagctggg agagaccctg ggaatgagca gagccgccat caacaagcac atccagacac
     3541 tgagagactg gggagtggac gtgttcaccg tgcctggcaa gggctacagc ctgcctgagc
     3601 ctatccagct gctgaacgcc aagcagatcc tgggacagct ggatggcgga agcgtggccg
     3661 tgctgcctgt gatcgactcc accaatcagt acctgctgga cagaatcgga gagctgaagt
     3721 ccggcgacgc ctgcatcgcc gagtaccagc aggctggcag aggaggcaga ggacggaagt
     3781 ggttcagccc attcggagcc aacctgtacc tgtccatgtt ctggagactg gagcagggac
     3841 ctgctgctgc catcggactg agtctggtga tcggaatcgt gatggccgag gtgctgagaa
     3901 agctgggagc cgacaaggtg agagtgaagt ggcctaatga cctgtacctc caggaccgca
     3961 agctggctgg catcctggtg gagctgacag gcaagacagg cgatgccgct cagatcgtga
     4021 tcggagccgg aatcaacatg gccatgagaa gagtggagga gagcgtggtg aaccagggct
     4081 ggatcaccct gcaggaggct ggcatcaacc tggaccggaa caccctggcc gccatgctga
     4141 tcagagagct gagagccgct ctggagctgt tcgagcagga gggactggct ccttacctga
     4201 gcagatggga gaagctggac aacttcatca acagacctgt gaagctgatc atcggcgaca
     4261 aggaaatctt cggcatctcc agaggaatcg acaagcaggg agctctgctg ctggagcagg
     4321 acggaatcat caagccctgg atgggcggag aaatctccct gagaagcgca gagaagggat
     4381 ccagcggcgg cggcagcggc ggcggcggca gcactgaata taaacttgtg gtagttggag
     4441 ctgatggcgt aggcaagagt gccttgacga tacagctaat tcagaatcat tttgtggacg
     4501 aatatgatcc aacaatagag gattcctaca ggaagcaagt agtaattgat ggagaaacct
     4561 gtctcttgga tattctcgac acagcaggtc aagaggagta cagtgcaatg agggaccagt
     4621 acatgaggac tggggagggc tttctttgtg tatttgccat aaataatact aaatcatttg
     4681 aagatattca ccattataga gaacaaatta aaagagttaa ggactctgaa gatgtaccta
     4741 tggtcctagt aggaaataaa tgtgatttgc cttccagaac agtagacaca aaacaggctc
     4801 aggacttagc aagaagttat ggaattcctt ttattgaaac atcagcaaag acaagacagg
     4861 gtgttgatga tgccttctat acattagttc gagaaattcg aaaacataaa gaaaagatga
     4921 gcaaagatgg taaaaagaag aaaaagaagt caaagacaaa gtgtgtaatt atgtaactcg
     4981 agtctagagg gcccgacgcg tctggccacc ggtcgcccct ctccctcccc cccccctaac
     5041 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     5101 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     5161 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     5221 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     5281 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     5341 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     5401 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     5461 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     5521 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     5581 cgataatacc atggccaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     5641 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     5701 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5761 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5821 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggct cgcgcatggc
     5881 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5941 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     6001 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgctggggt
     6061 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     6121 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     6181 gcccggtgcc tgagttaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     6241 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     6301 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     6361 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     6421 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     6481 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6541 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     6601 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6661 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6721 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6781 ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt
     6841 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     6901 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     6961 gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga
     7021 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     7081 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7141 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7201 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     7261 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg tttaaacccg
     7321 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     7381 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     7441 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     7501 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     7561 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     7621 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     7681 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     7741 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7801 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7861 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7921 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7981 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     8041 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     8101 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     8161 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     8221 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     8281 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     8341 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     8401 attttcggat ctgatcagca cgtgatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     8461 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     8521 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     8581 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     8641 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     8701 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     8761 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     8821 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     8881 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     8941 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     9001 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     9061 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     9121 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     9181 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     9241 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     9301 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     9361 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     9421 cccagcactc gtccgagggc aaaggaatag cacgtgctac gagatttcga ttccaccgcc
     9481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     9541 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     9601 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     9661 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     9721 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     9781 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     9841 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     9901 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     9961 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    10021 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    10081 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    10141 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    10201 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    10261 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    10321 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    10381 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    10441 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    10501 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    10561 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    10621 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    10681 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    10741 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    10801 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    10861 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    10921 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    10981 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    11041 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    11101 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    11161 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    11221 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    11281 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    11341 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    11401 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    11461 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    11521 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    11581 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    11641 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    11701 tgagcaaaaa caggaaggca aaatgccgca aaaaagagaa taagggcgac acggaaatgt
    11761 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    11821 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    11881 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc
    11941 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    12001 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
    12061 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
    12121 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt
    12181 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
