LOCUS       sequence-353281-        9679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing PAmCherry1-tagged KRas 4B G12D mutant, 
            with an N-terminal dimerization domain (DD)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9679)
  AUTHORS   Nan X, Tamguney TM, Collisson EA, Lin LJ, Pitt C, Galeas J, Lewis S,
            Gray JW, McCormick F, Chu S
  TITLE     Ras-GTP dimers activate the Mitogen-Activated Protein Kinase (MAPK) 
            pathway.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Jun 30;112(26):7996-8001.
  PUBMED    26080442
REFERENCE   2  (bases 1 to 9679)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2015-06-30"; volume: "112(26)"; pages: 
            "7996-8001"
COMMENT     Sequence Label: pLenti-TO/CMV-puro-DD-PAmCherry1-KRas G12D
FEATURES             Location/Qualifiers
     source          1..9679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(177..245,246..1037)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(395..414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1208..1796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1697..1716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1950..1967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2084..2105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2120..2125,2126..2143,2144..2150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2158..2174
                     /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)."
     primer_bind     2163..2185
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2182..2198
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2182..2198
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2217..2237
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2219..2237
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2265..2491
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2492..2672
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2719..2844
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3337..3570
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3755..3799
                     /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    4066..4182
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        4205..4508
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4509..4712
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4663..4683
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4714..4732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4735..4753
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4794..4818
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4851..5171
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             5178..5204
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5235..5942
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein"
                     /label=PAmCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5380..5398)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5648..5667
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(5967..5999,6000..6002,6003..6533)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             5967..6533
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(6559..6583)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6614..7202
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6667..6687)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7085..7096)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        7216..7704
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7251..7274)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7637..7656
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7725..8324
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7725..7744)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8221..8241
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             8462..8695
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8767..8888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8804..8823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8858..8877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8928..9063
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9083..9102)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9084..9102)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9112..9129)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9112..9128)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9121..9143)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(9270..9679,1..46)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9357..9376)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9567..9588
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
       61 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      721 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1321 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1861 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1921 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     1981 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2041 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     2101 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     2161 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     2221 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat
     2281 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
     2341 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
     2401 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
     2461 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
     2521 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     2581 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     2641 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     2701 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     2761 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     2821 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     2881 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     2941 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     3001 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     3061 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     3121 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     3181 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     3241 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     3301 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     3361 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     3421 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     3481 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     3541 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     3601 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     3661 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     3721 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     3781 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     3841 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     3901 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     3961 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     4021 cagatccatt cgattagtga acggatctcg acggtatcgg ttaactttta aaagaaaagg
     4081 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     4141 aactaaagaa ttacaaaaac aaattacaaa attcaaaatt ttatcgataa gcttgggagt
     4201 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4261 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4321 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4381 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4441 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4501 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4561 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4621 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4681 gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga gatctcccta
     4741 tcagtgatag agatcgtcga ctagtccagt gtggtggaat tctgcagata tcaacaagtt
     4801 tgtacaaaaa agcaggctcc gcggccgccc ccttcaccat ggcttctaga ggagtgcagg
     4861 tggagactat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg
     4921 tgcactacac cgggatgctt gaagatggaa agaaagttga ttcctcccgg gacagaaaca
     4981 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg
     5041 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg
     5101 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc
     5161 taaaactgga aactagttat ccgtacgacg taccagacta cgcaaagctt ctcgagctgc
     5221 aggagctcgg taccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     5281 tcatgcgctt caaggtgcat atggagggct ccgtgaacgg ccacgtgttc gagatcgagg
     5341 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     5401 gtggccccct gcccttcacc tgggacatcc tgtcccctca gttcatgtac ggctccaacg
     5461 cctacgtgaa gcaccccgcc gacatccccg actacttcaa gctgtccttc cccgagggct
     5521 tcaagtggga gcgcgtgatg aagttcgagg acggcggcgt ggtgaccgtg acccaggact
     5581 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     5641 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggccctctcc gagcggatgt
     5701 accccgagga cggcgccctg aagggcgagg tcaagccaag ggtcaagctg aaggacggcg
     5761 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     5821 gcgcctacaa cgtcaaccgc aagttggaca tcacctccca caacgaggac tacaccatcg
     5881 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     5941 agagatctca tatgccatgg gtcgacatga ctgaatataa acttgtggta gttggagctg
     6001 atggcgtagg caagagtgcc ttgacgatac agctaattca gaatcatttt gtggacgaat
     6061 atgatccaac aatagaggat tcctacagga agcaagtagt aattgatgga gaaacctgtc
     6121 tcttggatat tctcgacaca gcaggtcaag aggagtacag tgcaatgagg gaccagtaca
     6181 tgaggactgg ggagggcttt ctttgtgtat ttgccataaa taatactaaa tcatttgaag
     6241 atattcacca ttatagagaa caaattaaaa gagttaagga ctctgaagat gtacctatgg
     6301 tcctagtagg aaataaatgt gatttgcctt ctagaacagt agacacaaaa caggctcagg
     6361 acttagcaag aagttatgga attcctttta ttgaaacatc agcaaagaca agacagggtg
     6421 ttgatgatgc cttctataca ttagttcgag aaattcgaaa acataaagaa aagatgagca
     6481 aagatggtaa aaagaagaaa aagaagtcaa agacaaagtg tgtaattatg taaggatcct
     6541 aaaagggtgg gcgcgccgac ccagctttct tgtacaaagt ggttgatatc cagcacagtg
     6601 gcggccgctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     6661 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     6721 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     6781 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     6841 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     6901 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6961 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     7021 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     7081 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     7141 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     7201 gcctggaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt
     7261 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca
     7321 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta
     7381 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac
     7441 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga
     7501 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc
     7561 agaggctggg aaggggtggg tccgggggcg ggctcagggg gcggggcggg cgcccgaagg
     7621 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     7681 tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc
     7741 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     7801 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     7861 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     7921 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     7981 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     8041 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     8101 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     8161 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     8221 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     8281 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     8341 cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc tcgagggccc ggtaccttta
     8401 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     8461 ctggaagggc tagctcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     8521 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     8581 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     8641 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     8701 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     8761 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     8821 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     8881 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     8941 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     9001 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     9061 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     9121 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     9181 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     9241 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     9301 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     9361 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     9421 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     9481 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     9541 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     9601 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     9661 aaaaaatgag ctgatttaa
//
