LOCUS       sequence-421527-       12208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EFS-TiCas9-P2A-Puro. Tamoxifen-inducible SpyCas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12208)
  AUTHORS   Tan IL, Perez AR, Lew RJ, Sun X, Baldwin A, Zhu YK, Shah MM, Berger 
            MS, Doudna JA, Fellmann C
  TITLE     Targeting the non-coding genome and temozolomide signature enables 
            CRISPR-mediated glioma oncolysis.
  JOURNAL   Cell Rep. 2023 Nov 28;42(11):113339. doi: 
            10.1016/j.celrep.2023.113339. Epub 2023 Nov 2.
  PUBMED    37917583
REFERENCE   2  (bases 1 to 12208)
  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: "Cell Rep.";
            date: "2023-11-28"; volume: "42(11):113339. doi"; pages: " 
            10.1016/j.celrep.2023.113339. Epub 2023 Nov 2"
COMMENT     Sequence Label: pCF287_EFS-TiCas9-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..12208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        232..611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     766..786
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             829..1009
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1056..1181
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1674..1907
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2092..2136
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2434..2551
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3238..3449
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3468..3477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5853..8351
                     /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             8361..8417
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8418..9014
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8911..8931
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9030..9618
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9083..9103)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9501..9512)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9690..9923
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9949..9966)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9955..10179
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(10285..10302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10456..11044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10536..10555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11215..12006,12007..12075))
                     /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     11838..11857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12076..12180)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat tgacattgat
      241 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      301 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      361 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      481 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      541 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      601 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      661 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      721 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      781 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
      841 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      901 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      961 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     1021 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     1081 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1141 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1201 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1261 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1321 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1381 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1441 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1501 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1561 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1621 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1681 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1741 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1801 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1861 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1921 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1981 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     2041 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     2101 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     2161 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     2221 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     2281 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     2341 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     2401 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     2461 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     2521 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     2581 agtttggtta attaaggtac cgggaggtgg tccctgcagt tacgccaatg ataacccccg
     2641 ccagaaaaat cttagtagcc ttcccttttt gttttccgtg ccccaactcg gcggattgac
     2701 tcggcccctt ccggaaacac ccgaatcaac ttctagtcaa attattgttc acgccgcaat
     2761 gacccacccc tggcccgcgt ctgtggaact gacccctggt gtacaggaga gttcgctgct
     2821 gaaagtggtc ccaaaggggt actagttttt aagctcccaa ctccccctcc cccagcgtct
     2881 ggaggattcc acaccctcgc accgcagggg cgaggaagtg ggcggagtcc ggttttggcg
     2941 ccagccgctg aggctgccaa gcagaaaagc caccgctgag gagactccgg tcactgtcct
     3001 cgccccgcct cccccttccc tccccttggg gaccaccggg cgccacgccg cgaacggtaa
     3061 gtgccgcggt cgtcggcgcc tccgccctcc ccctagggcc ccaattccca gcgggcgcgg
     3121 cgcgcggccc cgcgcgcagc tcccggctcc ctcccccttc ggatgtggct tgagctgtag
     3181 gcgcggaggc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     3241 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     3301 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     3361 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     3421 ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc accatggaca
     3481 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
     3541 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     3601 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     3661 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     3721 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     3781 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     3841 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     3901 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     3961 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     4021 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     4081 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     4141 aaaatctgat cgcccagctg cccggcgcat ctccagcaaa cagcctggcc ttgtccctga
     4201 ccgccgacca gatggtcagt gccttgttgg atgctgagcc ccccatactc tattccgagt
     4261 atgatcctac cagacccttc agtgaggctt cgatgatggg cttactgacc aacctggcag
     4321 acagggagct ggttcacatg atcaactggg cgaagagggt gccaggcttt gtggatttga
     4381 ccctccatga tcaggcccac cttctggaat gtgcctggct agagatcctg atgattggac
     4441 tcgtctggcg cagcatggag catccagtga agctactgtt tgctcctaac ttgctcttgg
     4501 acaggaacca gggaaaatgt gtagagggca tggtggaaat cttcgacatg ctgctggcta
     4561 catcatctcg gttccgcatg atgaatctgc agggagagga gtttgtgtgc ctcaaatcta
     4621 ttattttgct taattctgga gtgtacacat ttctgtccag caccctgaag tctctggaag
     4681 agaaggacca tatccaccga gtcctggaca agatcacaga cactttgatc cacctgatgg
     4741 ccaaggcagg actgaccctg cagcagcagc accagcggct ggcccagctc ctcctcatcc
     4801 tctcccacat caggcacatg agtaacaaag gcatggagca tctgtacagc atgaagtgca
     4861 agaacgtggt gcccctctat gacctgctgc tggaggccgc cgacgcccac cgcctacatg
     4921 cgccacctgc atctccaggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     4981 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     5041 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     5101 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     5161 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     5221 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     5281 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     5341 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     5401 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     5461 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     5521 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     5581 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     5641 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     5701 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     5761 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     5821 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     5881 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     5941 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     6001 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     6061 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     6121 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     6181 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     6241 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     6301 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     6361 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     6421 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     6481 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     6541 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     6601 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     6661 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     6721 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
     6781 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     6841 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     6901 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     6961 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     7021 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     7081 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     7141 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     7201 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     7261 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     7321 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     7381 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
     7441 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     7501 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     7561 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     7621 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     7681 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
     7741 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     7801 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     7861 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
     7921 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
     7981 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
     8041 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
     8101 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
     8161 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
     8221 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
     8281 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
     8341 tgggaggcga cggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg
     8401 aagagaatcc tggaccgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     8461 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     8521 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     8581 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     8641 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     8701 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     8761 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg
     8821 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     8881 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     8941 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     9001 agcccggtgc ctgaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa
     9061 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     9121 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     9181 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     9241 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     9301 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     9361 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     9421 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     9481 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     9541 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     9601 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct
     9661 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     9721 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     9781 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     9841 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     9901 gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     9961 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
    10021 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
    10081 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
    10141 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
    10201 gccattaatg aaaagaacca gccattaatg aatcggccaa cgcgcgggga gaggcggttt
    10261 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    10321 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    10381 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    10441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    10501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    10561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    10621 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    10681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    10741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    10801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    10861 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    10921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    10981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    11041 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    11101 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
    11161 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
    11221 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
    11281 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
    11341 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
    11401 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
    11461 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    11521 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    11581 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
    11641 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
    11701 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    11761 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    11821 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    11881 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    11941 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    12001 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    12061 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    12121 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    12181 cacatttccc cgaaaagtgc cacctgac
//
