LOCUS       sequence-325841-       12516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduce Dox/Cre-inducible (TRE promoter followed by lox-stop-lox) 
            Cas9 cDNA plus Pten-targeting sgRNA into (ES) cells by 
            recombination-mediated cassette exchange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12516)
  TITLE     Karreth lab plasmids
REFERENCE   2  (bases 1 to 12516)
  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"
COMMENT     Sequence Label: cTRE-LSL-GFP-IRES-Cas9-sgPtenX1.1
FEATURES             Location/Qualifiers
     source          1..12516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..547
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(83..106)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     480..499
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    593..640
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    join(673..738,739..776)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    945..1079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1199..1333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1349..1597
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1349..1369
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1520..1539
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1619..1694
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    join(1742..1760,1761..1783,1784..1802,1803..1825,1826..1844,
                     1845..1867,1868..1886,1887..1909,1910..1928,1929..1951,
                     1952..1970,1971..1993,1994..2012)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2045..2083
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2080..2104
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(2149..2182)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          2720..2785
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2915..2935
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3360..3494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3536..3569)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3609..3618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3615..3617,3618..3620,3621..4334)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3921..3940)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4268..4289
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4342..4909
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4359..4911
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4503..4520)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4730..4749
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      4946..4955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6617..9115
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
                     SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
                     AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH
                     DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
                     NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
                     QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
                     VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
                     LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
                     GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
                     ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL
                     PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
                     RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP
                     SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
                     KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL
                     IHQSITGLYETRIDLSQLGGD"
     CDS             9119..9139
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          9183..9755
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(10083..10671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10163..10182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10842..11633,11634..11702))
                     /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     11465..11484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11703..11807)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11833..12288)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11970..11991)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12182..12201
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12415..12437
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12429..12446
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12430..12446
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12453..12472
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12453..12471
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(12504..12516,1..4)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(12504..12516,1..4)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gatcccccgg gctgcaggaa ttaattcgtc gacctcgaaa ttctaccggg taggggaggc
       61 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
      121 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
      181 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
      241 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
      301 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
      361 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
      421 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
      481 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
      541 cctttcgacc tgcatccatc tagatctcga tcgagcagct gaagcttatg gagaagttac
      601 tattccgaag ttcctattct ctagaaagta taggaacttc gatccccctc gatcccccgg
      661 gctgcagatc tgtagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg
      721 tccctttttt ttccacagct cgcggttgag gacaaactct tcgcggtctt tccagtgggg
      781 atcgacggta tcgataagct tgcagatctg cgactctaga ggatctgcga ctctagagga
      841 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
      901 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     1081 gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1141 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1321 tcatgtctgg atcgagttta ccatgcatga gggcctattt cccatgattc cttcatattt
     1381 gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa
     1441 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     1501 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     1561 cttggcttta tatatcttgt ggaaaggacg aaacaccgag atcgttagca gaaacaaagt
     1621 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     1681 caccgagtcg gtgctttttt gaattcgatg catcttatca tgtctggatc gagtttacca
     1741 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
     1801 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
     1861 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
     1921 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
     1981 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagctc ggtacccggg
     2041 tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca
     2101 gatcgcctgg agacgccatc cacgctcgag acctcttcga gggacctaat aacttcgtat
     2161 agcatacatt atacgaagtt atattaaggg ttccggatcc tcggggacac caaatatggc
     2221 gatctcggcc ttttcgtttc ttggagctgg gacatgtttg ccatcgatcc atctaccacc
     2281 agaacggccg ttagatctgc tgccaccgtt gtttccaccg aagaaaccac cgttgccgta
     2341 accaccacga cggttgttgc taaagaagct gccaccgcca cggccaccgt tgtagccgcc
     2401 gttgttgtta ttgtagttgc tactgttatt tctggcactt cttggttttc ctcttaagtg
     2461 aggaggaaca taaccattct cgttgttgtc gttgatgctt aaattttgca cttgttcgct
     2521 cagttcagcc ataatatgaa atgcttttct tgttgttctt acggaatacc acttgccacc
     2581 tatcaccaca actaactttt tcccgttcct ccatctcttt tatatttttt ttctcgatcg
     2641 agggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac
     2701 agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactact
     2761 gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat gggagcagtg
     2821 gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga
     2881 tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga
     2941 ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag
     3001 aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc tatacaagaa
     3061 aattatggaa aaatattctg taacctttat aagtaggcat aacagttata atcataacat
     3121 actgtttttt cttactccac acaggcatag agtgtctgct attaataact atgctcaaaa
     3181 attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt tgatgtatag
     3241 tgccttgact agagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3301 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3361 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3421 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3481 atcatgtctg gatctgacat ggtaagtaag cttgggctgc aggtcgaggg acctaataac
     3541 ttcgtatagc atacattata cgaagttata ttaagggttc cggatcctaa gcttgatgga
     3601 attctcgagc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3661 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc gagggcgagg
     3721 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     3781 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     3841 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3901 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3961 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     4021 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     4081 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggca
     4141 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     4201 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     4261 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     4321 agctgtacaa gtaatgaatt ttccgccccc cccccctaac gttactggcc gaagccgctt
     4381 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     4441 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     4501 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     4561 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     4621 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     4681 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     4741 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     4801 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     4861 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccctcg
     4921 tcgagtattg atcagtggat ccggtgccac catggactat aagrwssmcs mmrrrraswa
     4981 smrgrakswy grtatysayk rmrwmsmmgm wgmcgayaag awgkmcmgca wsggcctgga
     5041 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     5101 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     5161 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     5221 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     5281 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     5341 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     5401 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     5461 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     5521 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     5581 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     5641 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     5701 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     5761 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     5821 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     5881 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     5941 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     6001 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     6061 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     6121 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     6181 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     6241 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     6301 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     6361 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     6421 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     6481 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     6541 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     6601 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     6661 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     6721 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     6781 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     6841 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     6901 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     6961 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     7021 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     7081 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     7141 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     7201 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     7261 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     7321 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     7381 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     7441 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     7501 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     7561 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     7621 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     7681 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     7741 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     7801 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     7861 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     7921 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     7981 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     8041 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     8101 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     8161 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     8221 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     8281 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     8341 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     8401 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     8461 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     8521 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     8581 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     8641 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     8701 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     8761 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     8821 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     8881 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     8941 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     9001 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     9061 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacagccc
     9121 caagaagaag agaaaggtgg aggccagcta agaattcgca gtcgagcgtt gagaacttca
     9181 gggtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat
     9241 atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc
     9301 atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc
     9361 ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac
     9421 gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt
     9481 tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag agaacaattg
     9541 ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata
     9601 ttcttattgg tagaaacaac tacaccctgg tcatcatcct gcctttctct ttatggttac
     9661 aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg
     9721 ctaaccatgt tcatgccttc ttctctttcc tacagctcct gggcaacgtg ctggttgttg
     9781 tgctgtctca tcattttggc aaaggattca ctcctcaggt gcaggctgcc tatcagaagg
     9841 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatcgt tttccctctg
     9901 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     9961 aatttatttt cattgcaata gtgtgttgga atttctcgat cgctagcatc ctaggcgcgc
    10021 cacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
    10081 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
    10141 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
    10201 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
    10261 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
    10321 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
    10381 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
    10441 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
    10501 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
    10561 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
    10621 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
    10681 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
    10741 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
    10801 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
    10861 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
    10921 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
    10981 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
    11041 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
    11101 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
    11161 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
    11221 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
    11281 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
    11341 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
    11401 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
    11461 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
    11521 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
    11581 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
    11641 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    11701 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
    11761 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
    11821 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
    11881 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
    11941 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
    12001 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
    12061 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
    12121 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
    12181 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
    12241 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
    12301 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
    12361 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
    12421 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
    12481 agctccaccg cggtggcggc cgctctagaa ctagtg
//
