LOCUS       sequence-339457-       13124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA against mouse Mr1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13124)
  AUTHORS   Ruf B, Catania VV, Wabitsch S, Ma C, Diggs LP, Zhang Q, Heinrich B, 
            Subramanyam V, Cui LL, Pouzolles M, Evans CN, Chari R, Sakai S, Oh 
            S, Barry CE III, Barber DL, Greten TF
  TITLE     Activating Mucosal-Associated Invariant T Cells Induces a Broad 
            Antitumor Response.
  JOURNAL   Cancer Immunol Res. 2021 Jun 30. pii: 2326-6066.CIR-20-0925. doi: 
            10.1158/2326-6066.CIR-20-0925.
  PUBMED    34193462
REFERENCE   2  (bases 1 to 13124)
  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: "Cancer 
            Immunol Res. 2021 Jun 30. pii: 2326-6066.CIR-20-0925. doi: 
            10.1158/2326-6066.CIR-20-0925."
COMMENT     Sequence Label: pGMC00013
FEATURES             Location/Qualifiers
     source          1..13124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..53)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        225..604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..804
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     759..779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             822..1002
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1049..1174
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1667..1900
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2085..2129
                     /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    2427..2544
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2595..2843
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2595..2615
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2766..2785
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2865..2940
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3002..3213
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3232..3241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3238..7341
                     /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             3238..7341
                     /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             7342..7389
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7390..7413
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7423..7479
                     /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             7480..8187
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(7622..7640)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7890..7909
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    8203..8791
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8256..8276)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8674..8685)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8863..9096
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9122..9139)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9128..9352
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9398..9826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9485..9504)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9695..9716
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9835..9855)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9840..10169
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10020..10155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10082..10101
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10217..10264
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10283..10657
                     /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    10787..10908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10824..10843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10878..10897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10957..10973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10957..10973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10970..10992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10981..10997
                     /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(11005..11011,11012..11029,11030..11035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11050..11071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11188..11205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11359..11947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11439..11458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12118..12909,12910..12978))
                     /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     12741..12760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12979..13083)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
       61 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
      121 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      181 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      241 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      601 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      781 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
      841 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      901 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      961 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     1021 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     1081 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     1141 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1201 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1261 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1321 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1381 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1441 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1501 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1561 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1621 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1681 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1741 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1801 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1861 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1921 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1981 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2041 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2101 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2161 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2221 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2281 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2341 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2401 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2461 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2521 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2581 ttaattaagg taccgagggc ctatttccca tgattccttc atatttgcat atacgataca
     2641 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
     2701 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     2761 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     2821 tcttgtggaa aggacgaaac accgacactg tcgtatgtag tgatgtttta gagctagaaa
     2881 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     2941 ttttttgaat tcgctagcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
     3001 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3061 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3121 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3181 ctttttcgca acgggtttgc cgccagaaca caggaccggt tctagagcgc tgccaccatg
     3241 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     3301 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     3361 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     3421 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     3481 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     3541 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     3601 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     3661 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     3721 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     3781 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     3841 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     3901 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     3961 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     4021 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     4081 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     4141 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     4201 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     4261 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     4321 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     4381 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     4441 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     4501 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     4561 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     4621 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     4681 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     4741 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     4801 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     4861 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     4921 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     4981 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     5041 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     5101 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     5161 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     5221 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     5281 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     5341 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     5401 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     5461 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     5521 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     5581 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     5641 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     5701 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     5761 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     5821 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     5881 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     5941 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     6001 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     6061 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     6121 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     6181 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     6241 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     6301 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     6361 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     6421 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     6481 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     6541 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     6601 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     6661 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     6721 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     6781 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     6841 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     6901 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     6961 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     7021 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     7081 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     7141 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     7201 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     7261 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     7321 ctgtctcagc tgggaggcga caagcgacct gccgccacaa agaaggctgg acaggctaag
     7381 aagaagaaag attacaaaga cgatgacgat aagggatccg gcgcaacaaa cttctctctg
     7441 ctgaaacaag ccggagatgt cgaagagaat cctggaccgg tgagcaaggg cgaggaggat
     7501 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     7561 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     7621 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     7681 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     7741 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     7801 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     7861 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     7921 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     7981 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     8041 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     8101 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     8161 ggcggcatgg acgagctgta caagtaaacg cgttaagtcg acaatcaacc tctggattac
     8221 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     8281 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     8341 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     8401 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     8461 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     8521 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     8581 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     8641 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     8701 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     8761 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     8821 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     8881 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     8941 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     9001 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     9061 tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca
     9121 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     9181 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     9241 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     9301 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     9361 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     9421 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     9481 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     9541 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     9601 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     9661 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     9721 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     9781 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     9841 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     9901 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     9961 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
    10021 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
    10081 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
    10141 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
    10201 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
    10261 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
    10321 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
    10381 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
    10441 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
    10501 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
    10561 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
    10621 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
    10681 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
    10741 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
    10801 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
    10861 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
    10921 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
    10981 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
    11041 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
    11101 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
    11161 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
    11221 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
    11281 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
    11341 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
    11401 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
    11461 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
    11521 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
    11581 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
    11641 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
    11701 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
    11761 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
    11821 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
    11881 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
    11941 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
    12001 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
    12061 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
    12121 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
    12181 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
    12241 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
    12301 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
    12361 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
    12421 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
    12481 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
    12541 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
    12601 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
    12661 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
    12721 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
    12781 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
    12841 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
    12901 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
    12961 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
    13021 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
    13081 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgg
//
