LOCUS       sequence-366178-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting murine Msh2 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Povedano JM, Liou J, Wei D, Srivatsav A, Kim J, Xie Y, Nijhawan D, 
            McFadden DG
  TITLE     Engineering Forward Genetics into Cultured Cancer Cells for Chemical
            Target Identification.
  JOURNAL   Cell Chem Biol. 2019 Sep 19;26(9):1315-1321.e3. doi: 
            10.1016/j.chembiol.2019.06.006. Epub 2019 Jul 11.
  PUBMED    31303577
REFERENCE   2  (bases 1 to 13012)
  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.chembiol.2019.06.006"; journalName: "Cell Chem Biol"; 
            date: "2019-09-19- 19"; volume: "26"; issue: "9"; pages: 
            "1315-1321.e3"
COMMENT     Sequence Label: LentiCRISPR-V2_mMSH2
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..50)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        222..601
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..801
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     756..776
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             819..999
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1046..1171
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1664..1897
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2082..2126
                     /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    2424..2541
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2592..2840
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2592..2612
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2763..2782
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2861..2936
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2998..3209
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3228..3237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3234..7337
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3234..7337
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7338..7385
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7386..7409
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7419..7475
                     /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             7476..8072
                     /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     7969..7989
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8088..8676
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8141..8161)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8559..8570)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8748..8981
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9007..9024)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9013..9237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9283..9711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9370..9389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9580..9601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9720..9740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9725..10054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9905..10040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9967..9986
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10102..10149
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10168..10542
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10672..10793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10709..10728)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10763..10782
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10842..10858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10842..10858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10855..10877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10866..10882
                     /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(10890..10896,10897..10914,10915..10920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10935..10956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11073..11090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11244..11832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11324..11343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12003..12794,12795..12863))
                     /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     12626..12645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12864..12968)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
       61 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
      121 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      181 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      241 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      301 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      361 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      421 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      481 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      541 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      601 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      661 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      721 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      781 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
      841 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      901 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      961 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     1021 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     1081 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     1141 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     1201 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     1261 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     1321 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1381 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1441 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1501 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     1561 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     1621 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     1681 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     1741 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     1801 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     1861 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     1921 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     1981 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     2041 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     2101 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     2161 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     2221 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     2281 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     2341 acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga
     2401 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     2461 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     2521 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta
     2581 attaaggtac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     2641 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
     2701 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     2761 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     2821 tgtggaaagg acgaaacacc ggttaatacc ctgatacagt gttttagagc tagaaatagc
     2881 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     2941 ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     3001 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     3061 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     3121 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     3181 ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc accatggaca
     3241 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
     3301 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     3361 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     3421 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     3481 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     3541 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     3601 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     3661 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     3721 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     3781 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     3841 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     3901 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     3961 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     4021 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     4081 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     4141 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     4201 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     4261 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     4321 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     4381 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     4441 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     4501 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     4561 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     4621 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     4681 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     4741 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     4801 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     4861 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     4921 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     4981 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     5041 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     5101 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     5161 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     5221 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     5281 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     5341 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     5401 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     5461 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     5521 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     5581 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     5641 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     5701 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     5761 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     5821 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     5881 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     5941 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     6001 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     6061 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     6121 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     6181 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     6241 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     6301 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     6361 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     6421 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     6481 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     6541 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     6601 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     6661 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     6721 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     6781 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     6841 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     6901 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     6961 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     7021 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     7081 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     7141 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     7201 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     7261 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     7321 ctcagctggg aggcgacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga
     7381 agaaagatta caaagacgat gacgataagg gatccggcgc aacaaacttc tctctgctga
     7441 aacaagccgg agatgtcgaa gagaatcctg gaccgaccga gtacaagccc acggtgcgcc
     7501 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     7561 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     7621 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     7681 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     7741 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     7801 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag
     7861 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     7921 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     7981 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     8041 cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg
     8101 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     8161 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     8221 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     8281 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     8341 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     8401 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     8461 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     8521 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     8581 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     8641 agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta
     8701 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     8761 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     8821 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     8881 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     8941 atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct
     9001 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     9061 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     9121 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     9181 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     9241 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     9301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     9361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     9421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     9481 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     9541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     9601 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     9661 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     9721 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     9781 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     9841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     9901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     9961 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
    10021 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
    10081 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
    10141 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
    10201 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
    10261 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
    10321 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
    10381 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
    10441 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
    10501 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
    10561 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
    10621 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
    10681 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    10741 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    10801 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
    10861 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
    10921 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
    10981 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
    11041 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
    11101 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    11161 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    11221 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    11281 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    11341 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    11401 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    11461 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    11521 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    11581 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    11641 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    11701 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    11761 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    11821 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    11881 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
    11941 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
    12001 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
    12061 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
    12121 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
    12181 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    12241 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    12301 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    12361 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    12421 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    12481 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    12541 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    12601 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    12661 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    12721 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    12781 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    12841 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    12901 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    12961 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg ag
//
