LOCUS       sequence-466839-       12823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knockout Pnpt1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12823)
  AUTHORS   Miao R, Jiang C, Chang WY, Zhang H, An J, Ho F, Chen P, Zhang H, 
            Junqueira C, Amgalan D, Liang FG, Zhang J, Evavold CL, 
            Hafner-Bratkovic I, Zhang Z, Fontana P, Xia S, Waldeck-Weiermair M, 
            Pan Y, Michel T, Bar-Peled L, Wu H, Kagan JC, Kitsis RN, Zhang P, 
            Liu X, Lieberman J
  TITLE     Gasdermin D permeabilization of mitochondrial inner and outer 
            membranes accelerates and enhances pyroptosis.
  JOURNAL   Immunity. 2023 Nov 14;56(11):2523-2541.e8. doi: 
            10.1016/j.immuni.2023.10.004. Epub 2023 Nov 3.
  PUBMED    37924812
REFERENCE   2  (bases 1 to 12823)
  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.immuni.2023.10.004"; journalName: "Immunity"; date: 
            "2023-11-14- 14"; volume: "56"; issue: "11"; pages: "2523-2541.e8"
COMMENT     Sequence Label: lentiCRISPR v2-sgPnpt1-2#
FEATURES             Location/Qualifiers
     source          1..12823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..64
                     /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             65..463
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    488..1076
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(541..561)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(959..970)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1148..1381
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1407..1424)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1413..1637
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1683..2111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1770..1789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1980..2001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2120..2140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2125..2454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2305..2440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2367..2386
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2502..2549
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2568..2942
                     /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    3072..3193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3109..3128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3163..3182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3242..3258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3242..3258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3255..3277)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3266..3282
                     /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(3290..3296,3297..3314,3315..3320))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3335..3356
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3473..3490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3644..4232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3724..3743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4403..5194,5195..5263))
                     /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     5026..5045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5264..5368)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5443..5462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5634..6013
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6015..6213
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6168..6188
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6231..6411
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6458..6583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7076..7309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7494..7538
                     /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    7836..7953
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8004..8252
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8004..8024
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8175..8194
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8273..8348
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8410..8621
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8640..8649
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8646..12749
                     /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             8646..12749
                     /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             12750..12797
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12798..12821
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 atccggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
       61 accgatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
      121 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
      181 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
      241 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
      301 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
      361 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
      421 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagaattcg atatcaagct
      481 tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
      541 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
      601 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
      661 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
      721 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
      781 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
      841 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
      901 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
      961 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     1021 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     1081 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     1141 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     1201 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     1261 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     1321 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     1381 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1441 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1501 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1561 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     1621 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     1681 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1741 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1801 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1861 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1921 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1981 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2041 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2101 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2161 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2221 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2281 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2341 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2401 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2461 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     2521 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     2581 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     2641 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     2701 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     2761 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     2821 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     2881 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     2941 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3001 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3061 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3121 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3181 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3241 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3301 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3361 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3421 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3481 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3541 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3601 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3661 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3721 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3781 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3841 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3901 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3961 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4021 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4081 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4141 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4201 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4261 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4321 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4381 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4441 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4501 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4561 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4621 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4681 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4741 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4801 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4861 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4921 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4981 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5041 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5101 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5161 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5221 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5281 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5341 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     5401 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     5461 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     5521 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     5581 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     5641 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5701 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     5761 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     5821 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     5881 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     5941 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     6001 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     6061 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     6121 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     6181 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
     6241 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6301 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6361 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     6421 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     6481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     6541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     6601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     6661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     6721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     6781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     6841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     6901 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     6961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     7021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     7081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     7141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     7201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     7261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     7321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     7381 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     7441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     7501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     7561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     7621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     7681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     7741 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
     7801 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     7861 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     7921 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     7981 ccagtttggt taattaaggt accgagggcc tatttcccat gattccttca tatttgcata
     8041 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
     8101 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     8161 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     8221 ctttatatat cttgtggaaa ggacgaaaca ccgccttccc aattcatgcc gtgttttaga
     8281 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     8341 gtcggtgctt ttttgaattc gctagctagg tcttgaaagg agtgggaatt ggctccggtg
     8401 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     8461 gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     8521 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     8581 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggttc tagagcgctg
     8641 ccaccatgga caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg
     8701 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     8761 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     8821 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     8881 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     8941 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     9001 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     9061 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     9121 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     9181 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     9241 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     9301 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     9361 gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     9421 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     9481 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     9541 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     9601 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     9661 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     9721 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     9781 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     9841 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     9901 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     9961 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
    10021 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
    10081 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
    10141 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
    10201 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
    10261 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
    10321 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
    10381 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
    10441 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
    10501 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
    10561 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
    10621 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
    10681 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
    10741 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
    10801 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
    10861 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
    10921 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
    10981 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
    11041 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
    11101 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
    11161 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
    11221 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
    11281 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
    11341 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
    11401 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
    11461 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
    11521 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
    11581 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
    11641 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
    11701 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
    11761 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
    11821 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
    11881 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
    11941 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
    12001 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
    12061 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
    12121 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
    12181 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
    12241 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
    12301 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
    12361 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
    12421 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
    12481 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
    12541 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
    12601 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
    12661 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
    12721 ggatcgacct gtctcagctg ggaggcgaca agcgacctgc cgccacaaag aaggctggac
    12781 aggctaagaa gaagaaagat tacaaagacg atgacgataa ggg
//
