LOCUS       sequence-229004-       11295 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Flag-HA-6xHIS-N-Ras Q61K/E37A fusion protein in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11295)
  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 11295)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-FHH-NRASQ61K/E37A-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..11295
                     /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..3413
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3420..3446
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3456..3473
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4086..4658
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4253..4270)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4480..4499
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4661..5263
                     /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     5160..5180
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5270..5858
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5323..5343)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5741..5752)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6065..6298
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6399..6416)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6405..6629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6675..7103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6762..6781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6972..6993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7112..7132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7117..7446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7297..7432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7359..7378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7495..8520
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7495..8520
                     /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    8650..8771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8820..8836)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8820..8836)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8833..8855)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8844..8860
                     /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(8868..8874,8875..8892,8893..8898))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8913..8934
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9051..9068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9222..9810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9302..9321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9981..10772,10773..10841))
                     /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     10604..10623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10842..10946)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10994..11128
                     /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 attataaaga tgacgacgat aaagcaggtt
     3421 acccatacga tgtgcctgac tatgctgcag gttcacatca tcaccaccat catggatcca
     3481 ctgagtacaa actggtggtg gttggagcag gtggtgttgg gaaaagcgca ctgacaatcc
     3541 agctaatcca gaaccacttt gtagatgaat atgatcccac catagcggat tcttacagaa
     3601 aacaagtggt tatagatggt gaaacctgtt tgttggacat actggataca gctggaaaag
     3661 aagagtacag tgccatgaga gaccaataca tgaggacagg cgaaggcttc ctctgtgtat
     3721 ttgccatcaa taatagcaag tcatttgcgg atattaacct ctacagggag cagattaagc
     3781 gagtaaaaga ctcggatgat gtacctatgg tgctagtggg aaacaagtgt gatttgccaa
     3841 caaggacagt tgatacaaaa caagcccacg aactggccaa gagttacggg attccattca
     3901 ttgaaacctc agccaagacc agacagggtg ttgaagatgc tttttacaca ctggtaagag
     3961 aaatacgcca gtaccgaatg aaaaaactca acagcagtga tgatgggact cagggttgta
     4021 tgggattgcc atgtgtggtg atgtaactcg agtctagagg gcccgacgcg tctggccacc
     4081 ggtcgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     4141 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     4201 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     4261 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     4321 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     4381 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     4441 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     4501 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     4561 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     4621 cacggggacg tggttttcct ttgaaaaaca cgataatacc atggccaccg agtacaagcc
     4681 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     4741 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     4801 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4861 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4921 ggtgttcgcc gagatcggct cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4981 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5041 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5101 cggagtggag gcggccgagc gcgctggggt gcccgccttc ctggagacct ccgcgccccg
     5161 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5221 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagttaaca atcaacctct
     5281 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5341 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5401 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5461 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5521 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5581 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5641 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     5701 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5761 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5821 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga
     5881 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     5941 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     6001 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg
     6061 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     6121 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6181 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6241 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6301 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6361 agtgagaggc cttgacattg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     6421 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     6481 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6541 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6601 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6661 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     7021 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     7081 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     7141 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     7201 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     7261 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     7321 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     7381 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     7441 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgatgaaa
     7501 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     7561 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     7621 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     7681 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
     7741 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     7801 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     7861 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     7921 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     7981 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     8041 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     8101 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     8161 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     8221 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
     8281 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     8341 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     8401 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     8461 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
     8521 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     8581 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     8641 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     8701 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     8761 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     8821 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     8881 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     8941 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     9001 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     9061 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     9121 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     9181 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     9241 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     9301 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     9361 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     9421 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     9481 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     9541 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     9601 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     9661 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     9721 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     9781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     9841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     9901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     9961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    10021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    10081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    10141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    10201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    10261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    10321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    10381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    10441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    10501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    10561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    10621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    10681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    10741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    10801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    10861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    10921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
    10981 ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
    11041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    11101 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
    11161 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta
    11221 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac
    11281 tacaaactta gtagt
//
