Puro-Cas9 donor

LOCUS       Puro-Cas9 donor               11573 bp DNA     circular SYN 12-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11573)
  AUTHORS   Biofeng
  TITLE     Direct Submission
  JOURNAL   Exported Jul 12, 2023 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    899..920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        935..965
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    973..989
                     /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     997..1013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1034..1052
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    1087..1890
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    1897..1922
                     /label=SA
                     /note="splice acceptor site"
     CDS             1946..1999
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2009..2608
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    2646..2870
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2877..2958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3142..3166
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3192..3239)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(3240..7340)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7365..7385)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7392..7457)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      7457..7466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(7491..7515)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7532..7846)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    7606..7624
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7642..7660
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7677..7695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7713..7731
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7749..7767
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7784..7802
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7820..7838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    7863..8699
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(8734..8752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8759..8775)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8913..9215
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(10612..11472)
                     /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"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      901 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
      961 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     1021 acgccaagct cagaattaac cctcactaaa gggactagtc ctgcaggttt aaacgaattc
     1081 gccctttgct ttctctgacc agcattctct cccctgggcc tgtgccgctt tctgtctgca
     1141 gcttgtggcc tgggtcacct ctacggctgg cccagatcct tccctgccgc ctccttcagg
     1201 ttccgtcttc ctccactccc tcttcccctt gctctctgct gtgttgctgc ccaaggatgc
     1261 tctttccgga gcacttcctt ctcggcgctg caccacgtga tgtcctctga gcggatcctc
     1321 cccgtgtctg ggtcctctcc gggcatctct cctccctcac ccaaccccat gccgtcttca
     1381 ctcgctgggt tcccttttcc ttctccttct ggggcctgtg ccatctctcg tttcttagga
     1441 tggccttctc cgacggatgt ctcccttgcg tcccgcctcc ccttcttgta ggcctgcatc
     1501 atcaccgttt ttctggacaa ccccaaagta ccccgtctcc ctggctttag ccacctctcc
     1561 atcctcttgc tttctttgcc tggacacccc gttctcctgt ggattcgggt cacctctcac
     1621 tcctttcatt tgggcagctc ccctaccccc cttacctctc tagtctgtgc tagctcttcc
     1681 agccccctgt catggcatct tccaggggtc cgagagctca gctagtcttc ttcctccaac
     1741 ccgggcccct atgtccactt caggacagca tgtttgctgc ctccagggat cctgtgtccc
     1801 cgagctggga ccaccttata ttcccagggc cggttaatgt ggctctggtt ctgggtactt
     1861 ttatctgtcc cctccacccc acagtggggc aagcttctga cctcttctct tcctcccaca
     1921 gggcctcgag agatctggca gcggagaggg cagaggaagt cttctaacat gcggtgacgt
     1981 ggaggagaat cccggcccta ggctcgagat gaccgagtac aagcccacgg tgcgcctcgc
     2041 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     2101 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     2161 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     2221 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     2281 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     2341 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     2401 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     2461 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     2521 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     2581 gtgcatgacc cgcaagcccg gtgcctgatc tagagggccc gtttaaaccc gctgatcagc
     2641 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2701 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2761 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2821 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg gtcgaggcag
     2881 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2941 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3001 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     3061 gatcctctag agatatcgtc gacaagctta tcgatgcggc cgcgctagca cgcgtcagct
     3121 gactagagga tccccggatc aaccactttg tacaagaaag ctgggtcggc gcgcccaccc
     3181 ttgaattctt actttttctt ttttgcctgg ccggcctttt tcgtggccgc cggccttttg
     3241 tcgcctccca gctgagacag gtcgatccgt gtctcgtaca ggccggtgat gctctggtgg
     3301 atcagggtgg cgtccagcac ctctttggtg ctggtgtacc tcttccggtc gatggtggtg
     3361 tcaaagtact tgaaggcggc aggggctccc agattggtca gggtaaacag gtggatgata
     3421 ttctcggcct gctctctgat gggcttatcc cggtgcttgt tgtaggcgga cagcactttg
     3481 tccagattag cgtcggccag gatcactctc ttggagaact cgctgatctg ctcgatgatc
     3541 tcgtccaggt agtgcttgtg ctgttccaca aacagctgtt tctgctcatt atcctcgggg
     3601 gagcccttca gcttctcata gtggctggcc aggtacagga agttcacata tttggagggc
     3661 agggccagtt cgtttccctt ctgcagttcg ccggcagagg ccagcattct cttccggccg
     3721 ttttccagct cgaacaggga gtacttaggc agcttgatga tcaggtcctt tttcacttct
     3781 ttgtagccct tggcttccag aaagtcgatg ggattcttct cgaagctgct tctttccatg
     3841 atggtgatcc ccagcagctc tttcacactc ttcagtttct tggacttgcc cttttccact
     3901 ttggccacca ccagcacaga ataggccacg gtggggctgt cgaagccgcc gtacttctta
     3961 gggtcccagt ccttctttct ggcgatcagc ttatcgctgt tcctcttggg caggatagac
     4021 tctttgctga agccgcctgt ctgcacctcg gtctttttca cgatattcac ttggggcatg
     4081 ctcagcactt tccgcacggt ggcaaaatcc cggcccttat cccacacgat ctccccggtt
     4141 tcgccgtttg tctcgatcag aggccgcttc cggatctcgc cgttggccag ggtaatctcg
     4201 gtcttgaaaa agttcatgat gttgctgtag aagaagtact tggcggtagc cttgccgatt
     4261 tcctgctcgc tcttggcgat catcttccgc acgtcgtaca ccttgtagtc gccgtacacg
     4321 aactcgcttt ccagcttagg gtactttttg atcagggcgg ttcccacgac ggcgttcagg
     4381 taggcgtcgt gggcgtggtg gtagttgttg atctcgcgca ctttgtaaaa ctggaaatcc
     4441 ttccggaaat cggacaccag cttggacttc agggtgatca ctttcacttc ccggatcagc
     4501 ttgtcattct cgtcgtactt agtgttcatc cgggagtcca ggatctgtgc cacgtgcttt
     4561 gtgatctgcc gggtttccac cagctgtctc ttgatgaagc cggccttatc cagttcgctc
     4621 aggccgcctc tctcggcctt ggtcagattg tcgaactttc tctgggtaat cagcttggcg
     4681 ttcagcagct gccgccagta gttcttcatc ttcttcacga cctcttcgga gggcacgttg
     4741 tcgctcttgc cccggttctt gtcgcttctg gtcagcacct tgttgtcgat ggagtcgtcc
     4801 ttcagaaagc tctgaggcac gatatggtcc acatcgtagt cggacagccg gttgatgtcc
     4861 agttcctggt ccacgtacat atcccgccca ttctgcaggt agtacaggta cagcttctcg
     4921 ttctgcagct gggtgttttc cacggggtgt tctttcagga tctggctgcc cagctctttg
     4981 atgccctctt cgatccgctt cattctctcg cggctgttct tctgtccctt ctgggtggtc
     5041 tggttctctc tggccatttc gatcacgatg ttctcgggct tgtgccggcc catcactttc
     5101 acgagctcgt ccaccacctt cactgtctgc aggatgccct tcttaatggc ggggctgccg
     5161 gccagattgg caatgtgctc gtgcaggcta tcgccctggc cggacacctg ggctttctgg
     5221 atgtcctctt taaaggtcag gctgtcgtcg tggatcagct gcatgaagtt tctgttggcg
     5281 aagccgtcgg acttcaggaa atccaggatt gtcttgccgg actgcttgtc ccggatgccg
     5341 ttgatcagct tccggctcag cctgccccag ccggtgtatc tccgccgctt cagctgcttc
     5401 atcactttgt cgtcgaacag gtgggcatag gttttcagcc gttcctcgat catctctctg
     5461 tcctcaaaca gtgtcagggt cagcacgata tcttccagaa tgtcctcgtt ttcctcattg
     5521 tccaggaagt ccttgtcctt gataattttc agcagatcgt ggtatgtgcc cagggaggcg
     5581 ttgaaccgat cttccacgcc ggagatttcc acggagtcga agcactcgat tttcttgaag
     5641 tagtcctctt tcagctgctt cacggtcact ttccggttgg tcttgaacag caggtccacg
     5701 atggcctttt tctgctcgcc gctcaggaag gcgggctttc tcattccctc ggtcacgtat
     5761 ttcactttgg tcagctcgtt atacacggtg aagtactcgt acagcaggct gtgcttgggc
     5821 agcaccttct cgttgggcag gttcttatcg aagttggtca tccgctcgat gaagctctgg
     5881 gcggaagcgc ccttgtccac cacttcctcg aagttccagg gggtgatggt ttcctcgctc
     5941 tttctggtca tccaggcgaa tctgctgttt cccctggcca gagggcccac gtagtagggg
     6001 atgcggaagg tcaggatctt ctcgatcttt tcccggttgt ccttcaggaa tgggtaaaaa
     6061 tcttcctgcc gccgcagaat ggcgtgcagc tctcccaggt ggatctggtg ggggatgctg
     6121 ccgttgtcga aggtccgctg cttccgcagc aggtcctctc tgttcagctt cacgagcagt
     6181 tcctcggtgc cgtccatctt ttccaggatg ggcttgatga acttgtagaa ctcttcctgg
     6241 ctggctccgc cgtcaatgta gccggcgtag ccgttcttgc tctggtcgaa gaaaatctct
     6301 ttgtacttct caggcagctg ctgccgcacg agagctttca gcagggtcag gtcctggtgg
     6361 tgctcgtcgt atctcttgat catagaggcg ctcagggggg ccttggtgat ctcggtgttc
     6421 actctcagga tgtcgctcag caggatggcg tcggacaggt tcttggcggc cagaaacagg
     6481 tcggcgtact ggtcgccgat ctgggccagc aggttgtcca ggtcgtcgtc gtaggtgtcc
     6541 ttgctcagct gcagtttggc atcctcggcc aggtcgaagt tgctcttgaa gttgggggtc
     6601 aggcccaggc tcagggcaat caggtttccg aacaggccat tcttcttctc gccgggcagc
     6661 tgggcgatca gattttccag ccgtctgctc ttgctcagtc tggcagacag gatggccttg
     6721 gcgtccacgc cgctggcgtt gatggggttt tcctcgaaca gctggttgta ggtctgcacc
     6781 agctggatga acagcttgtc cacgtcgctg ttgtcggggt tcaggtcgcc ctcgatcagg
     6841 aagtggcccc ggaacttgat catgtgggcc agggccagat agatcagccg caggtcggcc
     6901 ttgtcggtgc tgtccaccag tttctttctc aggtggtaga tggtggggta cttctcgtgg
     6961 taggccacct cgtccacgat gttgccgaag atggggtgcc gctcgtgctt cttatcctct
     7021 tccaccagga aggactcttc cagtctgtgg aagaagctgt cgtccacctt ggccatctcg
     7081 ttgctgaaga tctcttgcag atagcagatc cggttcttcc gtctggtgta tcttcttctg
     7141 gcggttctct tcagccgggt ggcctcggct gtttcgccgc tgtcgaacag cagggctccg
     7201 atcaggttct tcttgatgct gtgccggtcg gtgttgccca gcaccttgaa tttcttgctg
     7261 ggcaccttgt actcgtcggt gatcacggcc cagcccacag agttggtgcc gatgtccagg
     7321 ccgatgctgt acttcttgtc ggctgctggg actccgtgga taccgacctt ccgcttcttc
     7381 tttggggcca tcttatcgtc atcgtctttg taatcaatat catgatcctt gtagtctccg
     7441 tcgtggtcct tatagtccat ggtggcaccg gtccaacctg aaaaccgcgg agcctgcttt
     7501 tttgtacaaa cttgttgatc ggaattctcc aggcgatctg acggttcact aaacgagctc
     7561 tgcttatata ggcctcccac cgtacacgcc tacctcgaca tacgttctct atcactgata
     7621 gggagtaaac tcgacatacg ttctctatca ctgataggga taaactcgac atacgttctc
     7681 tatcactgat agggagtaaa ctcgacatac gttctctatc actgataggg agtaaactcg
     7741 acatacgttc tctatcactg atagggagta aactcgacat cgttctctat cactgatagg
     7801 gagtaaactc gacatacgtt ctctatcact gatagggagt aaactcgaca gtactaagct
     7861 ttactaggga caggattggt gacagaaaag ccccatcctt aggcctcctc cttcctagtc
     7921 tcctgatatt gggtctaacc cccacctcct gttaggcaga ttccttatct ggtgacacac
     7981 ccccatttcc tggagccatc tctctccttg ccagaacctc taaggtttgc ttacgatgga
     8041 gccagagagg atcctgggag ggagagcttg gcagggggtg ggagggaagg gggggatgcg
     8101 tgacctgccc ggttctcagt ggccaccctg cgctaccctc tcccagaacc tgagctgctc
     8161 tgacgcggct gtctggtgcg tttcactgat cctggtgctg cagcttcctt acacttccca
     8221 agaggagaag cagtttggaa aaacaaaatc agaataagtt ggtcctgagt tctaactttg
     8281 gctcttcacc tttctagtcc ccaatttata ttgttcctcc gtgcgtcagt tttacctgtg
     8341 agataaggcc agtagccagc cccgtcctgg cagggctgtg gtgaggaggg gggtgtccgt
     8401 gtggaaaact ccctttgtga gaatggtgcg tcctaggtgt tcaccaggtc gtggccgcct
     8461 ctactccctt tctctttctc catccttctt tccttaaaga gtccccagtg ctatctggga
     8521 catattcctc cgcccagagc agggtcccgc ttccctaagg ccctgctctg ggcttctggg
     8581 tttgagtcct tggcaagccc aggagaggcg ctcaggcttc cctgtccccc ttcctcgtcc
     8641 accatctcat gcccctggct ctcctgcccc ttccctacag gggttcctgg ctctgctcta
     8701 agggcgaatt cgcggccgct aaattcaatt cgccctatag tgagtcgtat tacaattcac
     8761 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     8821 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     8881 cttcccaaca gttgcgcagc ctatacgtac ggcagtttaa ggtttacacc tataaaagag
     8941 agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg ccggggcgac
     9001 ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt
     9061 acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg
     9121 tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca
     9181 aaaacgccat taacctgatg ttctggggaa tataaatgtc aggcatgaga ttatcaaaaa
     9241 ggatcttcac ctagatcctt ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc
     9301 ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc
     9361 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
     9421 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     9481 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaagc tctgatcaag
     9541 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     9601 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     9661 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     9721 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     9781 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     9841 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     9901 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     9961 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
    10021 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
    10081 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatgca tggcgatgcc
    10141 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
    10201 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
    10261 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
    10321 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattattaa cgcttacaat
    10381 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catcaggtgg
    10441 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
    10501 tatgtatccg ctcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    10561 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    10621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    10681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    10741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    10801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    10861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    10921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    10981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    11041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    11101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    11161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    11221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    11281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    11341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    11401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    11461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    11521 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccg
//