LOCUS       sequence-6982-ex       19451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19451)
  AUTHORS   Jackson EL, Willis N, Mercer K, Bronson RT, Crowley D, Montoya R, 
            Jacks T, Tuveson DA
  TITLE     Analysis of lung tumor initiation and progression using conditional 
            expression of oncogenic K-ras.
  JOURNAL   Genes Dev. 2001 Dec 15. 15(24):3243-8.
  PUBMED    11751630
REFERENCE   2  (bases 1 to 19451)
  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: "Genes Dev. 
            2001 Dec 15. 15(24):3243-8."
COMMENT     Sequence Label: LSL K-ras G12D
FEATURES             Location/Qualifiers
     source          1..19451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3752..3785)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4326..4925)
                     /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(4409..4429)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4906..4925
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4969..5468)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     intron          6078..6143
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6273..6293
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6718..6839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     intron          6932..6997
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7127..7147
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7572..7693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     intron          7786..7851
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7981..8001
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8426..8547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     intron          8640..8705
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8835..8855
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9280..9401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      9426..9435
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(9466..9499)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     13000..13016
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        13048..13547
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             13558..14352
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(13612..13631)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     14222..14241
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    14354..14927
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    14377..14929
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(14521..14538)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     14748..14767
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     5'UTR           14933..14972
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             14990..16120
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGAAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(16118..16137)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    16162..16210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(17217..17233)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17218..17234)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(17263..17283)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(17263..17281)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(17302..17318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17302..17318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(17315..17337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    17326..17342
                     /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(17350..17356,17357..17374,17375..17380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    17395..17416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(17533..17550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(17704..18292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(17784..17803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(18463..19254,19255..19323))
                     /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     19086..19105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(19324..19428)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcccccgcg gccgcggcgg ctgaggcggc
      721 agcgctgtgg cggcggctga gacggcaggg gaaggcggcg gcggctcggc ccggagtccc
      781 gctcccgcgc catttcggac ccggagcgag cgcggcgcgg gcctgaaggc ggcggcggga
      841 gcctgaggcg cggcggctcc gcggcgcgga gagaggtagg gcgcgggcgg cgctcctggg
      901 ccgctgcccg gtgcggagct cggcgcggcc gggcggggcc gcggcgctga atgaagccgg
      961 ctcggggcgg cgggcgcggg gcgccgaggt cgggcggccg ccgtttcccg gacggggcgg
     1021 ctttccttcc tcctcccggc cgcggcgggc cctccgcgct cgcccggcct ccgcgcccgg
     1081 cctctgctgc acccggcatc cgcgcctggc gccccgcgcc cggctcaact tgtcgggcgc
     1141 gcgggctgtg cggggacgcc gcttcccctg gccgggctgc ggggccccgg gggaggtcgc
     1201 ggcgggaggt gctcggaccc ggtggccgga gggcgtcgtg gttctcggag gggaaaggcg
     1261 tccgccgcgg agttcgcgac cgctcccctc ccccacgcct gtaagggctg tgacattgcc
     1321 gcggccacaa aggaggaggt ggaggtaggg atggtgctgg cagaacaggt ggcccacgcc
     1381 ctgcgcggaa agcccgcgag cgacccgcac ggaagcagaa aagttaatgc ggcaagtctg
     1441 agtggctttg tgtcgtgaaa acacaggggg aacgaagggc gagagccgag gcaggggact
     1501 ctccccgagt ggagcgcagg cagctcggct cacagcgtca agtttgaatc gcctgctgcc
     1561 ctctcccctc agtcctgcgg tttgtccctt gccccttcca gactccagag ttgattgtag
     1621 cattcgggcc gacctctgaa atccccctag gcccatgtag tagctcaagt cctgagccag
     1681 ccccgttaaa gcaaggtgat catcaaggca aggacttaaa agcagaatgg catgctgtgt
     1741 ttgtcgtaga ggaatcatct gagaggaaat gttgaacaag tctgctcata ataatcactt
     1801 cgaaattgcc accaaatgac cagtgtcact cttccaccga gggcctgatt gctgacccct
     1861 catgtgatag acaatagaca tcagatacgc taggtatcca tatatttaat ttgaaactgt
     1921 agctattcac aagcatcctt ggttttaaga gatggcggat agtaatttaa tttttccact
     1981 ttggttggaa ttgagcccct ttattacctt ttgtaactgg gggtgggggt gggtgggtgg
     2041 gggaaccttt caggcttaag gtttttcttg agtaagactt cttttttttc tagtggactt
     2101 aattaaatat tactaagttt taaaaacccg acaataaatt ttattcagca aaagctacgg
     2161 tgtaaatgat tcaatatctg tccacagtat accctaaatt agccaacatt ttagtctgtg
     2221 ggaccccttt gcacctataa acaaattatt gagggtctca gaaatccttg aaggggggga
     2281 acatgccaat atttactata ttagaagtaa cacatttttt cagtttattt agaactagga
     2341 ataaagtgcc tgttggtaac atgaattcat tttttaatga aaaaataata aattttctag
     2401 aacaaagaat tagagtagct ttgttttaca cattaacaat ctgtttaatt gtgcttcata
     2461 ggacataaca gtattctttc atatctacat ctcatatctg catctgcatt ctctttgttg
     2521 tgattacagg gcctagagaa ccaactgtcc ttgtaatagt aagaaggcta aaaaacaatt
     2581 gctgtaaaaa cggttctaac ctcctaatcc tgttccctac cacacaaagt tttagggacc
     2641 caaaattgct cctgtacgct agccacactg tttgcgggag ggagaggcaa aggtcgcatt
     2701 tccatgtgat ttcttttcat ctatcccatt tgccatggtc tacatttgtt acaaaacttg
     2761 aaggcaaaca ggtaagttag gttagtagtg gctcacagga tggccccaga caagcagcca
     2821 gaaaggatag gtggggcaaa gtagggcagg agttggatga aggtctgata acaatggttt
     2881 tctgcccttg tggtcctgtt actgagggct tcaggcacag ggtaaggaag acagtgtgtt
     2941 ggaaaaacaa actgcatttt cccctcttgg tgcctgtgtg aacagttctt ggtggtcaat
     3001 aagtagactg tattttgtgt ttgtttacac tgtatatact tacattgacc tggaactatc
     3061 tattctataa caaaattgtg actttagtgc agttcttttg tatccatttg tctctttatc
     3121 tgcaataata tatgttctac aagggaaagg agataaagtg tatatattaa ttttatttca
     3181 aaaatgtttg gaaaagcaga ctaggaaaca acagtgttgt attcagggct gttaactgta
     3241 tacagtgcca gcacttagtg cacctgttgg gagtgcagtg tattataagc aaggcagaag
     3301 tcacagaggt tactgagaaa ctcttttatg ttctttacat tttttttctt gccaaaaata
     3361 gtctttatct ctgaatttca ttcaatttta tttccttgta cagaggtact gtttttagta
     3421 ctcaaaggat tatatgttaa aattgatctg atcctatacc aaatatttaa acccatttcc
     3481 cacagatagt ttatagatta aaatgccatc aaatattatc tttatttgct tcattgccta
     3541 aatatgtctt tccccagcac agtgcagttt tgacaccagc ttcggcttcc tattttgttg
     3601 ccctttattg cagaactgct ctgatggctt taaaacagtt ctctgcagtt gttggctcca
     3661 acacagatgt tcttaggcta cctaacttct aacttttaat atccagtcaa caaagaatac
     3721 cgcaagggta ggtgttggga tagctgtcga cataacttcg tatagcatac attatacgaa
     3781 gttatctcga ccagcttctg atggaattag aacttggcaa aacaatactg agaatgaagt
     3841 gtatgtggaa cagaggctgc tgatctcgtt cttcaggcta tgaaactgac acatttggaa
     3901 accacagtac ttagaaccac aaagtgggaa tcaagagaaa aacaatgatc ccacgagaga
     3961 tctatagatc tatagatcat gagtgggagg aatgagctgg cccttaattt ggttttgctt
     4021 gtttaaatta tgatatccaa ctatgaaaca ttatcataaa gcaatagtaa agagccttca
     4081 gtaaagagca ggcatttatc taatcccacc ccacccccac ccccgtagct ccaatccttc
     4141 cattcaaaat gtaggtactc tgttctcacc cttcttaaca aagtatgaca ggaaaaactt
     4201 ccattttagt ggacatcttt attgtttaat agatcatcaa tttctgcatc ccggggatct
     4261 gatatcatcg atgcatgggg tcgtgcgctc ctttcggtcg ggcgctgcgg gtcgtggggc
     4321 gggcgtcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg
     4381 acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag
     4441 gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg
     4501 gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac
     4561 gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc
     4621 cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg
     4681 acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg
     4741 ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg
     4801 tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg
     4861 gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg
     4921 gtcatggtaa gcttcagctg ctcgatcgag atctagatgg atgcaggtcg aaaggcccgg
     4981 agatgaggaa gaggagaaca gcgcggcaga cgtgcgcttt tgaagcgtgc agaatgccgg
     5041 gcctccggag gaccttcggg cgcccgcccc gcccctgagc ccgcccctga gcccgccccc
     5101 ggacccaccc cttcccagcc tctgagccca gaaagcgaag gagcaaagct gctattggcc
     5161 gctgccccaa aggcctaccc gcttccattg ctcagcggtg ctgtccatct gcacgagact
     5221 agtgagacgt gctacttcca tttgtcacgt cctgcacgac gcgagctgcg gggcgggggg
     5281 gaacttcctg actaggggag gagtagaagg tggcgcgaag gggccaccaa agaacggagc
     5341 cggttggcgc ctaccggtgg atgtggaatg tgtgcgaggc cagaggccac ttgtgtagcg
     5401 ccaagtgccc agcggggctg ctaaagcgca tgctccagac tgccttggga aaagcgcctc
     5461 ccctacccgg tagaatttcg aggtcgaagg gcgcagtagt ccagggtttc cttgatgatg
     5521 tcatacttat cctgtccctt ttttttccac agggatcctc ggggacacca aatatggcga
     5581 tctcggcctt ttcgtttctt ggagctggga catgtttgcc atcgatccat ctaccaccag
     5641 aacggccgtt agatctgctg ccaccgttgt ttccaccgaa gaaaccaccg ttgccgtaac
     5701 caccacgacg gttgttgcta aagaagctgc caccgccacg gccaccgttg tagccgccgt
     5761 tgttgttatt gtagttgctc atgttatttc tggcacttct tggttttcct cttaagtgag
     5821 gaggaacata accattctcg ttgttgtcgt tgatgcttaa attttgcact tgttcgctca
     5881 gttcagccat aatatgaaat gcttttcttg ttgttcttac ggaataccac ttgccaccta
     5941 tcaccacaac taactttttc ccgttcctcc atctctttta tatttttttt ctcgatcgag
     6001 ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
     6061 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga
     6121 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt
     6181 ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg
     6241 aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc
     6301 ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa
     6361 ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa
     6421 ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat cataacatac
     6481 tgttttttct tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat
     6541 tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg
     6601 ccttgactag agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     6661 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     6721 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6781 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6841 catgtctggt cgagggatct ttgtgaagga accttacttc tgtggtgtga cataattgga
     6901 caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg
     6961 tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga
     7021 atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc
     7081 atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag
     7141 aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt
     7201 gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact
     7261 gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta
     7321 taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa
     7381 ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata
     7441 tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt
     7501 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     7561 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7621 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     7681 tcaatgtatc ttatcatgtc tggtcgaggg atctttgtga aggaacctta cttctgtggt
     7741 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
     7801 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
     7861 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
     7921 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
     7981 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
     8041 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
     8101 gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt
     8161 aggcataaca gttataatca taacatactg ttttttctta ctccacacag gcatagagtg
     8221 tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg
     8281 gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac
     8341 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     8401 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     8461 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8521 tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg agggatcttt gtgaaggaac
     8581 cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa agctctaagg
     8641 taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat
     8701 tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc tttaatgagg
     8761 aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact gctgactctc
     8821 aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac tttccttcag
     8881 aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct tgctttgcta
     8941 tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa aaatattctg
     9001 taacctttat aagtaggcat aacagttata atcataacat actgtttttt cttactccac
     9061 acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc tttagctttt
     9121 taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact agagatcata
     9181 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     9241 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     9301 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     9361 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gaattccgca
     9421 agctagccac catggcttga gtaagtctgc aggtcgaggg acctaataac ttcgtatagc
     9481 atacattata cgaagttatg tcgacaagct catgcgggtg tgtccacagg gtatagcgta
     9541 ctatgcagaa tatttgtact gagtgaagtc atgatacatt cctttgagag ccattagctg
     9601 ctacaaaaca gtaatctggc tgtttagatc aacaagctaa atgatagaag atgaaagtac
     9661 tggtttccat gtatttttac taagtgttga tgagaaagtt gtaagtgact tacaggttac
     9721 tctgtacatc tgtagtcact gaattcggaa tatcttagag tcttacacac aaaggtgagt
     9781 gttaaaatat tgataaagtt tttgataatc ttgtgtgaga catgttctaa tttagttgta
     9841 ttttattatt tttattgtaa ggcctgctga aaatgactga gtataagctt gtggtggttg
     9901 gagctgatgg cgtaggcaag agcgccttga cgatacagct aattcagaat cactttgtgg
     9961 atgagtacga ccctacgata gaggtaacgc tgctctacag tctgcgtgcg cttgtaaagg
    10021 acggcagcca gccgctttga aaaagatatc atttttatat ttattagaaa attatattga
    10081 aagttatttc agttatatgt gatgtccttt agttccaagg ctttaaactg ggtgttaggg
    10141 aaccataggt gcaagaaact ccacttatca tgagagctca ccacagagaa taaggtccaa
    10201 gatgccatcc gtgctttgta tttgcatttt gataaataaa tttcatgatt ttgtgagagt
    10261 taagaatata gctttgcctt tgatctattc ctgaaaggtt tgttgttgtt ttatgggttt
    10321 gttgttattg tgttgttgtt tggtgtcttt taggcagggt cttactatgc agccctgacc
    10381 ggcctagaac tcaactatgt agaccagact agctctgaat tctcagagat ccatctacct
    10441 gcctctgctg cccagatgct gcgattaaag atgtacacca ccatgtacag tgaaagttac
    10501 tcttttgaaa acactttatt gttatagatt acttggaatg ttgatgtgat gagaaaattt
    10561 gattatacta ggaaaataga atcacttgat ggttgagctt tttttttaag acttatttat
    10621 tttatgtaag tgagtacact gcttctctct tcagacacac cagaagaggg catcagatcc
    10681 cattaagatg gctgtgagcc accatgtggt tgctgggaat tgaactcagg acctctggaa
    10741 aggcagtcag tgctcttttt ttgtttaaag atttatttat ttttatgaat acagtgtcac
    10801 tgtcttcata cacacaagag gaaagcatca gatcccatta cagatgattg cgagccacca
    10861 tgtggttgct gggaattgaa ctcaggacct ttggaagagc agcccatgct cttaaccact
    10921 gagccatctc tctagccccg atggttaaaa tttttaagct taatacagtt tgatgagtaa
    10981 gatcttcagg attattttca gacaattttc atttcataga atcttaacca attaaaggcc
    11041 aactgctaag aaatagaatt gatacctaga agaggtcttg aaaaatctgg tagtacctcc
    11101 tattgctgtc agtaggttta ggttttccta accaaagaag agtgtattta acctccgaga
    11161 ttagcaggaa gggcttttcg ttttttgttt gtttgtttgt ttggttggtt ggttggtttt
    11221 ggttttggtt ttggttttgg ttttttagag acagggtttc tttgtatagc cctggctgtc
    11281 ctggaactca ctctgtagac caggctggcc tcgagctcag aaatccacct gcttctgcct
    11341 cccgagtgct gggattaaag gcgtgcgcta ccacgcccgg ccaaggaagg gcttttcaca
    11401 aggtaattgt tccagtattt atgtgtagtc taaatatatt aaatttaata tcagtgaggt
    11461 gtgaaataaa tgttactttg cctttgaaat aacctattgt tagattattt ttaggcatct
    11521 ttactgcagt atgatgatat cccaatcttt agcttttcct ctttgttcta atgtgccatt
    11581 ttcagttctt tgataggttc ttatgtatcc caagatggct cttcaggtag ctgagaccta
    11641 gcgtgcaatc ctgatcatgc agcttccact ccttgagtga cagggctcca ttacaggctt
    11701 ttatgcctcc atgctcattc taggctgtgc tggggaatcg aatccacgcc ctctgacttg
    11761 ctgggcaatc ttctactctc caagctacat ccttacttca tgtaccaatt taaaaagaag
    11821 aaaaaaaaag acttgtgtag aattttgtag tgcttcagaa tccatagcta tgaaatctca
    11881 agtaggtcac agtgccctgg cttgggtctc tttctgtctt gaagaaacag tgtgctgggt
    11941 tgggctttga gggcagtggc gagcttggga aggtgattat ataacttaga atattgaagg
    12001 ttgattaaaa aattgttagg aagtgtcaaa taatgttagt agtagtatac cacccttcaa
    12061 aagatggtac ttagatataa aaatgacttt tatttgcttg tcaagtttaa caagttacat
    12121 cagtgacgta gagctcctct gtactagtct tcatacactg atttttgaca ggtcagctca
    12181 gtgctgtaat gtagttctgc tttttccttc cagaaagatc aaaaaggtca tccattttga
    12241 ctggtattct aaatgcaagg ttacttggca tgttatacca tttaaatcac atgtaaaagg
    12301 aagttttaat agaatgcatg tgaaatgtga acgttttcct gggatttgga agatgctttc
    12361 agcagaagtt aaccttcatt ttccttgaaa atctgagggg ttctattgtg cttcctcctg
    12421 ttagttcaga aggcatcccg aatatctaca gcatgacgtc agagctgtag gagcagggta
    12481 aaatagagct taggaagcct ttagttcacg tttgctaaaa tgtaatatac acctaacaat
    12541 tcatcatgta tgaagaaggt agaaaagtct ataaatttaa atttctaacc ttgattatga
    12601 tttctgccat gccagaagac ttgactctgg ggagcactga taacctgtga actttgaccc
    12661 tgacattccc ccaggtagtt tagcagctgg agaaaagtct ataccttctc tcacaatctg
    12721 tggtgagtct gaggaaactg acatggtcct ctcttggtgt ggtttagctg taagaagggc
    12781 aaaagagcga aagagattta attgactaaa aatgccaact tcctacatgt acaaaaagtt
    12841 tcaattgaag aaaagtgcca gagagaaaat gagagcatgg aagctgggtc tgggggcaga
    12901 tgctactcga tactgggagg cagaaggatt tccagtttaa tatctgcctg ggttacagag
    12961 caggtgcaag gtcagcctgc acagcttagt gagaccctgt ctagaactag tggatccccc
    13021 gggctgcagg aattcgataa ttctaccggg taggggaggc gcttttccca aggcagtctg
    13081 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
    13141 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
    13201 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
    13261 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
    13321 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
    13381 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
    13441 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
    13501 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagccatg
    13561 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
    13621 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
    13681 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
    13741 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
    13801 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
    13861 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
    13921 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
    13981 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
    14041 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
    14101 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
    14161 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
    14221 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
    14281 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
    14341 gagttcttct gagcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
    14401 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
    14461 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
    14521 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
    14581 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
    14641 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
    14701 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
    14761 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
    14821 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
    14881 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat cggatcttgg
    14941 tggcgtgaaa ctcccgcacc tcttcggcaa gcgccttgta gaagcgcgta tggcttcgta
    15001 cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg gccatagcaa
    15061 ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc gcctggagca
    15121 gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg ggaaaaccac
    15181 caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg tacccgagcc
    15241 gatgacttac tggcgggtgt tgggggcttc cgagacaatc gcgaacatct acaccacaca
    15301 acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa tgacaagcgc
    15361 ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc ctcatatcgg
    15421 gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct tcgaccgcca
    15481 tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca gcatgacccc
    15541 ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca caaacatcgt
    15601 gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc agcgccccgg
    15661 cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttatgggc tgcttgccaa
    15721 tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac agctttcggg
    15781 ggcggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac gaccccatat
    15841 cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca acggcgacct
    15901 gtataacgtg tttgcctggg ctttggacgt cttggccaaa cgcctccgtc ccatgcacgt
    15961 ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc tgcaacttac
    16021 ctccgggatg gtccagaccc acgtcaccac cccaggctcc ataccgacga tctgcgacct
    16081 ggcgcgcacg tttgcccggg agatggggga ggctaactga aacacggaag gagacaatac
    16141 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
    16201 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
    16261 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
    16321 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caagccctgc catagccacg
    16381 ggccccgtgg gttagggacg gggtccccca tggggaatgg tttatggttc gtgggggtta
    16441 ttattttggg cgttgcgtgg ggtcaggtcc acgactggac tgagcagaca gacccatggt
    16501 ttttggatgg cctgggcatg gaccgcatgt actggcgcga cacgaacacc gggcgtctgt
    16561 ggctgccaaa cacccccgac ccccaaaaac caccgcgcgg atttctggcg ccgccggacg
    16621 aactaaacct gactacggca tctctgcccc ttcttcgctg gtacgaggag cgcttttgtt
    16681 ttgtattggt caccacggcc gagtttccgc gggaccccgg ccaggacctg cagaaattga
    16741 tgatctatta aacaataaag atgtccacta aaatggaagt ttttcctgtc atactttgtt
    16801 aagaagggtg agaacagagt acctacattt tgaatggaag gattggagct acgggggtgg
    16861 gggtggggtg ggattagata aatgcctgct ctttactgaa ggctctttac tattgcttta
    16921 tgataatgtt tcatagttgg atatcataat ttaaacaagc aaaaccaaat taagggccag
    16981 ctcattcctc ccactcatga tctatagatc tatagatctc tcgtgggatc attgtttttc
    17041 tcttgattcc cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca gtttcatagc
    17101 ctgaagaacg agatcagcag cctctgttcc cacatacact tcattctcag tattgttttg
    17161 ccaagttcta attccatcag aagctgcggg ctgcaggaat tcgatatcaa gcttatcgat
    17221 accgtcgacc tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat
    17281 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    17341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    17401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    17461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    17521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    17581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    17641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    17701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    17761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    17821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    17881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    17941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    18001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    18061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    18121 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
    18181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    18241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    18301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    18361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
    18421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
    18481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
    18541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
    18601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    18661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    18721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    18781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    18841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    18901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    18961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    19021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    19081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    19141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    19201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    19261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    19321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    19381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    19441 aaaagtgcca c
//
