LOCUS       sequence-68867-e        9287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one plasmid targeting tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9287)
  AUTHORS   Hou Z, Zhang Y, Propson NE, Howden SE, Chu LF, Sontheimer EJ, 
            Thomson JA
  TITLE     Efficient genome engineering in human pluripotent stem cells using 
            Cas9 from Neisseria meningitidis.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Aug 12.
  PUBMED    23940360
REFERENCE   2  (bases 1 to 9287)
  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. 2013 Aug 12."
COMMENT     Sequence Label: 
            pSimpleII-U6-tracr-U6-crRNA(tdTomato)-NLS-NmCas9-HA-NLS(s)
FEATURES             Location/Qualifiers
     source          1..9287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(65..313)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     351..369
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(388..406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        complement(448..540)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     promoter        complement(540..788)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(853..872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1022..2195
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1253..2186
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2143..2163
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2201..2210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2213..2233
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2234..5476
                     /codon_start=1
                     /product="Cas9 endonuclease from the Neisseria meningitidis
                     Type II CRISPR/Cas system (Esvelt et al., 2013)"
                     /label=NmCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="AAFKPNSINYILGLDIGIASVGWAMVEIDEEENPIRLIDLGVRVF
                     ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRTRRLLKREGVLQAANFDENGLIKS
                     LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAG
                     NAHALQTGDFRTPAELALNKFEKESGHIRNQRSDYSHTFSRKDLQAELILLFEKQKEFG
                     NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
                     LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
                     AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
                     DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
                     EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
                     IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
                     LNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEFK
                     ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
                     FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
                     GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTLEKLRTLLAEK
                     LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
                     EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
                     GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
                     IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
                     IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     CDS             5477..5503
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5510..5536
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     primer_bind     complement(5552..5569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5558..5782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5971..7084
                     /label=mini-oriP
                     /note="internally truncated variant of the Epstein-Barr 
                     virus oriP replication origin (Tanaka et al., 1999)"
     primer_bind     complement(7130..7146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7130..7146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7143..7165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7154..7170
                     /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(7178..7184,7185..7202,7203..7208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7223..7244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7361..7378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7532..8120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7612..7631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8291..9082,9083..9151))
                     /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     8914..8933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9152..9256)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtttaaacaa aaaaacgaaa tgagaaaggg agctacaaca tgtcggcggg gtgcttcacg
       61 taccggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag
      121 ttacggtaag catatgatag tccattttaa aacataattt taaaactgca aactacccaa
      181 gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat
      241 taattctaat tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg
      301 gaaataggcc ctcttcctgc ccgaccttag atccgaatcc atatcgatat gctagttatt
      361 gctcagcggt ggcagcagcc aactcagcct atagtgagtc gtattaattt cgcgggatgt
      421 ttaaacgcgg ccgcaaaaaa accgaaataa acgatgcccc ttaaagcaga agctttaagg
      481 ggcagagcgt tgcggcacat cttttcagac ggccttattg tagcaacggt tctcatttcg
      541 gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg
      601 gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat
      661 tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt
      721 ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat catgggaaat
      781 aggccctctt cctgcccgac cttgacgtcg acggatcggg agatctcccg atcccctatg
      841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
      901 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
      961 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcgg
     1021 aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     1081 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     1141 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     1201 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     1261 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     1321 tacttccacc tggctccagt acgtgattct tgatcccgag ctggagccag gggcgggcct
     1381 tgcgctttag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
     1441 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     1501 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     1561 aatgcgggcc aggatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     1621 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     1681 atcggacggg ggtcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
     1741 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
     1801 cgtgagcgga aagatggccg cttcccggcc ctgctccagg gggctcaaaa tggaggacgc
     1861 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaggggcc tttccgtcct
     1921 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     1981 tctggagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     2041 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     2101 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     2161 ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacc gccaccatgg tgcctaagaa
     2221 gaagagaaag gtggctgcct tcaaacctaa ttcaatcaac tacatcctcg gcctcgatat
     2281 cggcatcgca tccgtcggct gggcgatggt agaaattgac gaagaagaaa accccatccg
     2341 cctgattgat ttgggcgtgc gcgtatttga gcgtgccgaa gtaccgaaaa caggcgactc
     2401 ccttgccatg gcaaggcgtt tggcgcgcag tgttcgccgc ctgacccgcc gtcgcgccca
     2461 ccgcctgctt cggacccgcc gcctattgaa acgcgaaggc gtattacaag ccgccaattt
     2521 tgacgaaaac ggcttgatta aatccttacc gaatacacca tggcaacttc gcgcagccgc
     2581 attagaccgc aaactgacgc ctttagagtg gtcggcagtc ttgttgcatt taatcaaaca
     2641 tcgcggctat ttatcgcaac ggaaaaacga gggcgaaact gccgataagg agcttggcgc
     2701 tttgcttaaa ggcgtagccg gcaatgccca tgccttacag acaggcgatt tccgcacacc
     2761 ggccgaattg gctttaaata aatttgagaa agaaagcggc catatccgca atcagcgcag
     2821 cgattattcg catacgttca gccgcaaaga tttacaggcg gagctgattt tgctgtttga
     2881 aaaacaaaaa gaatttggca atccgcatgt ttcaggcggc cttaaagaag gtattgaaac
     2941 cctactgatg acgcaacgcc ctgccctgtc cggcgatgcc gttcaaaaaa tgttggggca
     3001 ttgcaccttc gaaccggcag agccgaaagc cgctaaaaac acctacacag ccgaacgttt
     3061 catctggctg accaagctga acaacctgcg tattttagag caaggcagcg agcggccatt
     3121 gaccgatacc gaacgcgcca cgcttatgga cgagccatac agaaaatcca aactgactta
     3181 cgcacaagcc cgtaagctgc tgggtttaga agataccgcc tttttcaaag gcttgcgcta
     3241 tggtaaagac aatgccgaag cctcaacatt gatggaaatg aaggcctacc atgccatcag
     3301 ccgtgcactg gaaaaagaag gattgaaaga caaaaaatcc ccattaaacc tttctcccga
     3361 attacaagac gaaatcggca cggcattctc cctgttcaaa accgatgaag acattacagg
     3421 ccgtctgaaa gaccgtatac agcccgaaat cttagaagcg ctgttgaaac acatcagctt
     3481 cgataagttc gtccaaattt ccttgaaagc attgcgccga attgtgcctc taatggaaca
     3541 aggcaaacgt tacgatgaag cctgcgccga aatctacgga gaccattacg gcaagaagaa
     3601 tacggaagaa aagatttatc tgccgccgat tcccgccgac gaaatccgca accccgtcgt
     3661 cttgcgcgcc ttatctcaag cacgtaaggt cattaacggc gtggtacgcc gttacggctc
     3721 cccagctcgt atccatattg aaactgcaag ggaagtaggt aaatcgttta aagaccgcaa
     3781 agaaattgag aaacgccaag aagaaaaccg caaagaccgg gaaaaagccg ccgccaaatt
     3841 ccgagagtat ttccccaatt ttgtcggaga acccaaatcc aaagatattc tgaaactgcg
     3901 cctgtacgag caacaacacg gcaaatgcct gtattcgggc aaagaaatca acttaggccg
     3961 tctgaacgaa aaaggctatg tcgaaatcga ccatgccctg ccgttctcgc gcacatggga
     4021 cgacagtttc aacaataaag tactggtatt gggcagcgaa aaccaaaaca aaggcaatca
     4081 aaccccttac gaatacttca acggcaaaga caacagccgc gaatggcagg aatttaaagc
     4141 gcgtgtcgaa accagccgtt tcccgcgcag taaaaaacaa cggattctgc tgcaaaaatt
     4201 cgatgaagac ggctttaaag aacgcaatct gaacgacacg cgctacgtca accgtttcct
     4261 gtgtcaattt gttgccgacc gtatgcggct gacaggtaaa ggcaagaaac gtgtctttgc
     4321 atccaacgga caaattacca atctgttgcg cggcttttgg ggattgcgca aagtgcgtgc
     4381 ggaaaacgac cgccatcacg ccttggacgc cgtcgtcgtt gcctgctcga ccgttgccat
     4441 gcagcagaaa attacccgtt ttgtacgcta taaagagatg aacgcgtttg acggtaaaac
     4501 catagacaaa gaaacaggag aagtgctgca tcaaaaaaca cacttcccac aaccttggga
     4561 atttttcgca caagaagtca tgattcgcgt cttcggcaaa ccggacggca aacccgaatt
     4621 cgaagaagcc gataccctag aaaaactgcg cacgttgctt gccgaaaaat tatcatctcg
     4681 ccccgaagcc gtacacgaat acgttacgcc actgtttgtt tcacgcgcgc ccaatcggaa
     4741 gatgagcggg caagggcata tggagaccgt caaatccgcc aaacgactgg acgaaggcgt
     4801 cagcgtgttg cgcgtaccgc tgacacagtt aaaactgaaa gacttggaaa aaatggtcaa
     4861 tcgggagcgc gaacctaagc tatacgaagc actgaaagca cggctggaag cacataaaga
     4921 cgatcctgcc aaagcctttg ccgagccgtt ttacaaatac gataaagcag gcaaccgcac
     4981 ccaacaggta aaagccgtac gcgtagagca agtacagaaa accggcgtat gggtgcgcaa
     5041 ccataacggt attgccgaca acgcaaccat ggtgcgcgta gatgtgtttg agaaaggcga
     5101 caagtattat ctggtaccga tttacagttg gcaggtagcg aaagggattt tgccggatag
     5161 ggctgttgta caaggaaaag atgaagaaga ttggcaactt attgatgata gtttcaactt
     5221 taaattctca ttacacccta atgatttagt cgaggttata acaaaaaaag ctagaatgtt
     5281 tggttacttt gccagctgcc atcgaggcac aggtaatatc aatatacgca ttcatgatct
     5341 tgatcataaa attggcaaaa atggaatact ggaaggtatc ggcgtcaaaa ccgccctttc
     5401 attccaaaaa taccaaattg acgaactggg caaagaaatc agaccatgcc gtctgaaaaa
     5461 acgcccgcct gtccgttacc catacgatgt tccagattac gctgcagctc cagcagcgaa
     5521 gaaaaagaag ctggattaac tcgctgatca gcctcgactg tgccttctag ttgccagcca
     5581 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5641 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5701 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     5761 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgaccgat
     5821 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     5881 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc ctggccgggg
     5941 tcccccggaa actcggccgt ggtgaccatg caggaaaagg acaagcagcg aaaattcacg
     6001 cccccttggg aggtggcggc atatgcaaag gatagcactc ccactctact actgggtatc
     6061 atatgctgac tgtatatgca tgaggatagc atatgctacc cggatacaga ttaggatagc
     6121 atatactacc cagatataga ttaggatagc atatgctacc cagatataga ttaggatagc
     6181 ctatgctacc cagatataaa ttaggatagc atatactacc cagatataga ttaggatagc
     6241 atatgctacc cagatataga ttaggatagc ctatgctacc cagatataga ttaggatagc
     6301 atatgctacc cagatataga ttaggatagc atatgctatc cagatatttg ggtagtatat
     6361 gctacccaga tataaattag gatagcatat actaccctaa tctctattag gatagcatat
     6421 gctacccgga tacagattag gatagcatat actacccaga tatagattag gatagcatat
     6481 gctacccaga tatagattag gatagcctat gctacccaga tataaattag gatagcatat
     6541 actacccaga tatagattag gatagcatat gctacccaga tatagattag gatagcctat
     6601 gctacccaga tatagattag gatagcatat gctatccaga tatttgggta gtatatgcta
     6661 cccatggcaa cattagccca ccgtgctctc agcgacctcg tgaatatgag gaccaacaac
     6721 cctgtgcttg gcgctcaggc gcaagtgtgt gtaatttgtc ctccagatcg cagcaatcgc
     6781 gcccctatct tggcccgccc acctacttat gcaggtattc cccggggtgc cattagtggt
     6841 tttgtgggca agtggtttga ccgcagtggt tagcggggtt acaatcagcc aagttattac
     6901 acccttattt tacagtccaa aaccgcaggg cggcgtgtgg gggctgacgc gatcgctgtt
     6961 ccttaggacc cttttactaa ccctaattcg atagcatatg cttcccgttg ggtaacatat
     7021 gctattgaat tagggttagt ctggatagta tatactacta cccgggaagc atatgctacc
     7081 cgtttagggt tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     7141 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     7201 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7261 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7321 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7381 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7441 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7501 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7561 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7621 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7681 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     7741 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7801 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7861 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7921 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7981 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     8041 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     8101 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8161 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8221 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8281 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8341 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8401 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8461 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8521 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8581 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8641 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8701 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8761 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8821 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8881 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8941 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     9001 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     9061 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9121 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     9181 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9241 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
