LOCUS       sequence-230448-       12239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To clone sgRNA for SaCas9 (SauCas9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12239)
  AUTHORS   Lu B, Klingbeil O, Tarumoto Y, Somerville TDD, Huang YH, Wei Y, Wai 
            DC, Low JKK, Milazzo JP, Wu XS, Cao Z, Yan X, Demerdash OE, Huang G,
            Mackay JP, Kinney JB, Shi J, Vakoc CR
  TITLE     A Transcription Factor Addiction in Leukemia Imposed by the MLL 
            Promoter Sequence.
  JOURNAL   Cancer Cell. 2018 Dec 10;34(6):970-981.e8. doi: 
            10.1016/j.ccell.2018.10.015. Epub 2018 Nov 29.
  PUBMED    30503706
REFERENCE   2  (bases 1 to 12239)
  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"; doi: 
            "10.1016/j.ccell.2018.10.015"; journalName: "Cancer Cell"; date: 
            "2018-12-10- 10"; volume: "34"; issue: "6"; pages: "970-981.e8"
COMMENT     Sequence Label: Lenti_SaCRISPR_GFP
FEATURES             Location/Qualifiers
     source          1..12239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      232..241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             241..264
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             271..291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             316..3471
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             3472..3519
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3535..3591
                     /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             join(3592..3594,3595..3597,3598..4311)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3592..3594,3595..3597,3598..4308)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3637..3658)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3898..3917)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4245..4266
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4327..4915
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4380..4400)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4798..4809)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4987..5220
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5298..5419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5335..5354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5389..5408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5459..5594
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5614..5633)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5615..5633)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5643..5660)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5643..5659)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5652..5674)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5801..6256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5888..5907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6098..6119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6282..6386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6387..6455,6456..7247)
                     /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     complement(6605..6624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7418..8006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7907..7926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8160..8177
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8294..8315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8330..8335,8336..8353,8354..8360)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8368..8384
                     /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     8373..8395
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8392..8408
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8392..8408
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8427..8447
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8429..8447
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8475..8701
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8702..8882
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8929..9054
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9547..9780
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_RNA        complement(9842..9917)
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     promoter        complement(11799..12047)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(11857..11876)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(12027..12047)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    12054..12171
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
       61 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      181 tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg
      241 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
      301 ggagtcccag cagccaagcg gaactacatc ctgggcctgg acatcggcat caccagcgtg
      361 ggctacggca tcatcgacta cgagacacgg gacgtgatcg atgccggcgt gcggctgttc
      421 aaagaggcca acgtggaaaa caacgagggc aggcggagca agagaggcgc cagaaggctg
      481 aagcggcgga ggcggcatag aatccagaga gtgaagaagc tgctgttcga ctacaacctg
      541 ctgaccgacc acagcgagct gagcggcatc aacccctacg aggccagagt gaagggcctg
      601 agccagaagc tgagcgagga agagttctct gccgccctgc tgcacctggc caagagaaga
      661 ggcgtgcaca acgtgaacga ggtggaagag gacaccggca acgagctgtc caccaaagag
      721 cagatcagcc ggaacagcaa ggccctggaa gagaaatacg tggccgaact gcagctggaa
      781 cggctgaaga aagacggcga agtgcggggc agcatcaaca gattcaagac cagcgactac
      841 gtgaaagaag ccaaacagct gctgaaggtg cagaaggcct accaccagct ggaccagagc
      901 ttcatcgaca cctacatcga cctgctggaa acccggcgga cctactatga gggacctggc
      961 gagggcagcc ccttcggctg gaaggacatc aaagaatggt acgagatgct gatgggccac
     1021 tgcacctact tccccgagga actgcggagc gtgaagtacg cctacaacgc cgacctgtac
     1081 aacgccctga acgacctgaa caatctcgtg atcaccaggg acgagaacga gaagctggaa
     1141 tattacgaga agttccagat catcgagaac gtgttcaagc agaagaagaa gcccaccctg
     1201 aagcagatcg ccaaagaaat cctcgtgaac gaagaggata ttaagggcta cagagtgacc
     1261 agcaccggca agcccgagtt caccaacctg aaggtgtacc acgacatcaa ggacattacc
     1321 gcccggaaag agattattga gaacgccgag ctgctggatc agattgccaa gatcctgacc
     1381 atctaccaga gcagcgagga catccaggaa gaactgacca atctgaactc cgagctgacc
     1441 caggaagaga tcgagcagat ctctaatctg aagggctata ccggcaccca caacctgagc
     1501 ctgaaggcca tcaacctgat cctggacgag ctgtggcaca ccaacgacaa ccagatcgct
     1561 atcttcaacc ggctgaagct ggtgcccaag aaggtggacc tgtcccagca gaaagagatc
     1621 cccaccaccc tggtggacga cttcatcctg agccccgtcg tgaagagaag cttcatccag
     1681 agcatcaaag tgatcaacgc catcatcaag aagtacggcc tgcccaacga catcattatc
     1741 gagctggccc gcgagaagaa ctccaaggac gcccagaaaa tgatcaacga gatgcagaag
     1801 cggaaccggc agaccaacga gcggatcgag gaaatcatcc ggaccaccgg caaagagaac
     1861 gccaagtacc tgatcgagaa gatcaagctg cacgacatgc aggaaggcaa gtgcctgtac
     1921 agcctggaag ccatccctct ggaagatctg ctgaacaacc ccttcaacta tgaggtggac
     1981 cacatcatcc ccagaagcgt gtccttcgac aacagcttca acaacaaggt gctcgtgaag
     2041 caggaagaaa acagcaagaa gggcaaccgg accccattcc agtacctgag cagcagcgac
     2101 agcaagatca gctacgaaac cttcaagaag cacatcctga atctggccaa gggcaagggc
     2161 agaatcagca agaccaagaa agagtatctg ctggaagaac gggacatcaa caggttctcc
     2221 gtgcagaaag acttcatcaa ccggaacctg gtggatacca gatacgccac cagaggcctg
     2281 atgaacctgc tgcggagcta cttcagagtg aacaacctgg acgtgaaagt gaagtccatc
     2341 aatggcggct tcaccagctt tctgcggcgg aagtggaagt ttaagaaaga gcggaacaag
     2401 gggtacaagc accacgccga ggacgccctg atcattgcca acgccgattt catcttcaaa
     2461 gagtggaaga aactggacaa ggccaaaaaa gtgatggaaa accagatgtt cgaggaaaag
     2521 caggccgaga gcatgcccga gatcgaaacc gagcaggagt acaaagagat cttcatcacc
     2581 ccccaccaga tcaagcacat taaggacttc aaggactaca agtacagcca ccgggtggac
     2641 aagaagccta atagagagct gattaacgac accctgtact ccacccggaa ggacgacaag
     2701 ggcaacaccc tgatcgtgaa caatctgaac ggcctgtacg acaaggacaa tgacaagctg
     2761 aaaaagctga tcaacaagag ccccgaaaag ctgctgatgt accaccacga cccccagacc
     2821 taccagaaac tgaagctgat tatggaacag tacggcgacg agaagaatcc cctgtacaag
     2881 tactacgagg aaaccgggaa ctacctgacc aagtactcca aaaaggacaa cggccccgtg
     2941 atcaagaaga ttaagtatta cggcaacaaa ctgaacgccc atctggacat caccgacgac
     3001 taccccaaca gcagaaacaa ggtcgtgaag ctgtccctga agccctacag attcgacgtg
     3061 tacctggaca atggcgtgta caagttcgtg accgtgaaga atctggatgt gatcaaaaaa
     3121 gaaaactact acgaagtgaa tagcaagtgc tatgaggaag ctaagaagct gaagaagatc
     3181 agcaaccagg ccgagtttat cgcctccttc tacaacaacg atctgatcaa gatcaacggc
     3241 gagctgtata gagtgatcgg cgtgaacaac gacctgctga accggatcga agtgaacatg
     3301 atcgacatca cctaccgcga gtacctggaa aacatgaacg acaagaggcc ccccaggatc
     3361 attaagacaa tcgcctccaa gacccagagc attaagaagt acagcacaga cattctgggc
     3421 aacctgtatg aagtgaaatc taagaagcac cctcagatca tcaaaaaggg caagcgtcct
     3481 gctgctacta agaaagctgg tcaagctaag aaaaagaaag ctagcggcag cggcgccacc
     3541 aacttcagcc tgctgaagca ggccggcgac gtggaggaga accccggccc catggtgagc
     3601 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3661 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     3721 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3781 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3841 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3901 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3961 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     4021 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     4081 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     4141 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     4201 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     4261 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta gacgcgttaa
     4321 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4381 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4441 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4501 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4561 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4621 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4681 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4741 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4801 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4861 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
     4921 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     4981 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     5041 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     5101 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     5161 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     5221 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     5281 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5341 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5401 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     5461 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5521 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5581 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     5641 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5701 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5761 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     5821 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5881 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5941 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6001 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6061 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6121 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6181 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6241 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6301 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6361 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6421 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6481 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6541 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6601 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6661 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6721 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6781 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6841 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6901 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6961 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7021 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7081 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7141 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7201 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7261 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7321 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7381 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7441 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7501 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7561 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7621 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7681 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7741 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7801 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7861 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7921 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7981 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8041 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     8101 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     8161 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     8221 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     8281 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     8341 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     8401 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     8461 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     8521 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     8581 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     8641 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     8701 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     8761 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     8821 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     8881 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     8941 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     9001 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     9061 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     9121 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     9181 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     9241 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     9301 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     9361 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     9421 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     9481 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     9541 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     9601 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     9661 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     9721 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     9781 ggggatttgg ggttgctctg gaaaactcgc catttgtctc aagatctaga attcaaaaaa
     9841 atctcgccaa caagttgacg agataaacac ggcattttgc cttgtttaag tagattctgt
     9901 ttccagagta cttaaacaga gacgtacaaa aaagagcaag aagctaaaaa agatttaaaa
     9961 attattttta gcgcagttaa tggaacagga actaaattta ccccaaaaat attacgtgaa
    10021 tcaggatata acgttattga ggttgaagag catgcatttg aagatgaaac atttaaaaat
    10081 gttgtaaatc caaatccaga atttgatcct gcatgaaaaa taccgcttga atatggtatt
    10141 aaacatgatg cagatattat tattatgaat gacccagatg ctgacagatt tggaatggca
    10201 ataaaacatg atggtcattt tgtaagatta gatggaaatc aaacaggacc aattttaatt
    10261 gattgaaaat tatcaaatct aaaacgctta aatagcattc caaaaaatcc ggctctatat
    10321 tcaagttttg taacaagtga tttgggtgat agaatcgctc atgaaaaata tggagttaat
    10381 attgtaaaaa ctttaactgg atttaaatga atgggtagag aaattgctaa agaagaagat
    10441 aacggattaa attttgtttt tgcttatgaa gaaagttatg gatatgtaat tgatgactca
    10501 gctagagata aagatggaat acaagcttct atattaatag cagaggctgc ttgattttat
    10561 aaaaaacaaa ataaaacatt agtagactat ttagaagatt tatttaaaga aatgggtgca
    10621 tattacactt tcactttaaa cttgaatttt aaaccagaag aaaagaaatt aaaaattgaa
    10681 ccattaatga aatcattgag agcaacaccc ttaactcaaa ttgctggact taaagttgtt
    10741 aatgttgaag actacatcga tggaatgtat aatatgccag gacaagactt actaaaattt
    10801 tatttagaag ataagtcatg atttgctgtt cgcccaagtg gaactgaacc taaactaaaa
    10861 atttatttta taggtgttgg tgaatctgtt caaaacgcta aagttaaagt agacgaaatt
    10921 attaaagaat taaaattaaa aatgaatata taggagaaaa aatgaaacta aacaaatata
    10981 tagatcacac attattaaaa caagatgcta cgaaagctga aattaaacaa ttatgtgatg
    11041 aagcaattga atttgatttt gcaacagttt gtgttaattc atattgaaca agctattgta
    11101 aagaattatt aaaaggcaca aatgtaggaa taacaaatgt tgtaggtttt cctctaggtg
    11161 catgcacaac agctacaaaa gcattcgaag tttctgaagc aattaaagat ggtgcaacag
    11221 aaattgatat ggtattaaat attggtgcat taaaagacaa aaattatgaa ttagttttag
    11281 aagacatgaa agctgtaaaa aaagcagctg gatcacatgt tgttaaatgt attatggaaa
    11341 attgtttatt aacaaaagaa gaaatcatga aagcttgtga aatagctgtt gaagctggat
    11401 tagaatttgt taaaacatca acaggatttt caaaatcagg tgcaacattt gaagatgtta
    11461 aactaatgaa gtcagttgtt aaagacaatg ctttagttaa agcagctggt ggagttagaa
    11521 catttgaaga tgctcaaaaa atgattgaag caggagctga ccgcttagga acaagtggtg
    11581 gagtagctat tattaaaggt gaagaaaaca acgcgagtta ctaaaactag cgttttttta
    11641 ttttgctcat ttttattaaa agtttgcaaa aaggaacata aaaattctaa ttattgatac
    11701 taaagttatt aaaaagaaga ttttggttga ttttataaag gtcatagaat ataatatttt
    11761 agcatgtgta ttttgtgtgc tcatttacaa ccgtctccgg tgtttcgtcc tttccacaag
    11821 atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg atagtccatt
    11881 ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct acgtcacgta
    11941 ttttgtacta atatctttgt gtttacagtc aaattaattc taattatctc tctaacagcc
    12001 ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctccca caattttaaa
    12061 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
    12121 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
    12181 tacagggaca gcagagatcc actttggcgc cggctcgagt ggctccggtg cccgtcagt
//
