LOCUS       sequence-170102-       12560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dNmeCas9-3XsfGFP and telomere-targeting Nme sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12560)
  AUTHORS   Pawluk A, Amrani N, Zhang Y, Garcia B, Hidalgo-Reyes Y, Lee J, 
            Edraki A, Shah M, Sontheimer EJ, Maxwell KL, Davidson AR
  TITLE     Naturally Occurring Off-Switches for CRISPR-Cas9.
  JOURNAL   Cell. 2016 Dec 15;167(7):1829-1838.e9. doi: 
            10.1016/j.cell.2016.11.017. Epub 2016 Dec 8.
  PUBMED    27984730
REFERENCE   2  (bases 1 to 12560)
  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.cell.2016.11.017"; journalName: "Cell"; date: 
            "2016-12-15- 15"; volume: "167"; issue: "7"; pages: "1829-1838.e9"
COMMENT     Sequence Label: pEJS476-pHAGE-TO-Nme dCas9
            3XGFP-SgRNA/Telomer...
FEATURES             Location/Qualifiers
     source          1..12560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2299..2602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2603..2806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2757..2777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2808..2826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2857..2881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2974..2983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2986..3006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3019..3060,3061..3063,3064..4773,4774..4779,4780..4845,
                     4846..4848,4849..6261)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Neisseria meningitidis Type II 
                     CRISPR/Cas system"
                     /label=NmdCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D16A, D587A, 
                     H588A, and N611A"
                     /translation="AAFKPNPINYILGLAIGIASVGWAMVEIDEDENPICLIDLGVRVF
                     ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRARRLLKREGVLQAADFDENGLIKS
                     LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAD
                     NAHALQTGDFRTPAELALNKFEKESGHIRNQRGDYSHTFSRKDLQAELILLFEKQKEFG
                     NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
                     LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
                     AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
                     DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
                     EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
                     IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
                     LNEKGYVEIAAALPFSRTWDDSFNNKVLVLGSEAQNKGNQTPYEYFNGKDNSREWQEFK
                     ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
                     FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
                     GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTPEKLRTLLAEK
                     LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
                     EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
                     GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
                     IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
                     IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     regulatory      5848..5857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6280..6300
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6313..7026
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             7045..7758
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             7777..8490
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    8512..9100
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8565..8585)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8983..8994)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9175..9408
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        9504..9752
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9504..9524
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9675..9694
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9806..9898
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     primer_bind     complement(9965..9983)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10051..10155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10156..10224,10225..11016)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10374..10393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11187..11775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11676..11695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11929..11946
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12186..12207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12222..12227,12228..12245,12246..12252)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12260..12276
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     12265..12287
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12284..12300
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12284..12300
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12350..12369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag
     2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
     2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
     2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2941 cctaccggtt aaggtgaacg cgtgcaggct ggcgccacca tggcccctaa aaagaaacgc
     3001 aaggtcgaat tcggaagcgc cgccttcaag cccaacccca tcaactacat cctgggcctg
     3061 gccatcggca tcgccagcgt gggctgggcc atggtggaga tcgacgagga cgagaacccc
     3121 atctgcctga tcgacctggg tgtgcgcgtg ttcgagcgcg ctgaggtgcc caagactggt
     3181 gacagtctgg ctatggctcg ccggcttgct cgctctgttc ggcgccttac tcgccggcgc
     3241 gctcaccgcc ttctgcgcgc tcgccgcctg ctgaagcgcg agggtgtgct gcaggctgcc
     3301 gacttcgacg agaacggcct gatcaagagc ctgcccaaca ctccttggca gctgcgcgct
     3361 gccgctctgg accgcaagct gactcctctg gagtggagcg ccgtgctgct gcacctgatc
     3421 aagcaccgcg gctacctgag ccagcgcaag aacgagggcg agaccgccga caaggagctg
     3481 ggtgctctgc tgaagggcgt ggccgacaac gcccacgccc tgcagactgg tgacttccgc
     3541 actcctgctg agctggccct gaacaagttc gagaaggaga gcggccacat ccgcaaccag
     3601 cgcggcgact acagccacac cttcagccgc aaggacctgc aggccgagct gatcctgctg
     3661 ttcgagaagc agaaggagtt cggcaacccc cacgtgagcg gcggcctgaa ggagggcatc
     3721 gagaccctgc tgatgaccca gcgccccgcc ctgagcggcg acgccgtgca gaagatgctg
     3781 ggccactgca ccttcgagcc agccgagccc aaggccgcca agaacaccta caccgccgag
     3841 cgcttcatct ggctgaccaa gctgaacaac ctgcgcatcc tggagcaggg cagcgagcgc
     3901 cccctgaccg acaccgagcg cgccaccctg atggacgagc cctaccgcaa gagcaagctg
     3961 acctacgccc aggcccgcaa gctgctgggt ctggaggaca ccgccttctt caagggcctg
     4021 cgctacggca aggacaacgc cgaggccagc accctgatgg agatgaaggc ctaccacgcc
     4081 atcagccgcg ccctggagaa ggagggcctg aaggacaaga agagtcctct gaacctgagc
     4141 cccgagctgc aggacgagat cggcaccgcc ttcagcctgt tcaagaccga cgaggacatc
     4201 accggccgcc tgaaggaccg catccagccc gagatcctgg aggccctgct gaagcacatc
     4261 agcttcgaca agttcgtgca gatcagcctg aaggccctgc gccgcatcgt gcccctgatg
     4321 gagcagggca agcgctacga cgaggcctgc gccgagatct acggcgacca ctacggcaag
     4381 aagaacaccg aggagaagat ctacctgcct cctatccccg ccgacgagat ccgcaacccc
     4441 gtggtgctgc gcgccctgag ccaggcccgc aaggtgatca acggcgtggt gcgccgctac
     4501 ggcagccccg cccgcatcca catcgagacc gcccgcgagg tgggcaagag cttcaaggac
     4561 cgcaaggaga tcgagaagcg ccaggaggag aaccgcaagg accgcgagaa ggccgccgcc
     4621 aagttccgcg agtacttccc caacttcgtg ggcgagccca agagcaagga catcctgaag
     4681 ctgcgcctgt acgagcagca gcacggcaag tgcctgtaca gcggcaagga gatcaacctg
     4741 ggccgcctga acgagaaggg ctacgtggag atcgccgctg ccctgccctt cagccgcacc
     4801 tgggacgaca gcttcaacaa caaggtgctg gtgctgggca gcgaggctca gaacaagggc
     4861 aaccagaccc cctacgagta cttcaacggc aaggacaaca gccgcgagtg gcaggagttc
     4921 aaggcccgcg tggagaccag ccgcttcccc cgcagcaaga agcagcgcat cctgctgcag
     4981 aagttcgacg aggacggctt caaggagcgc aacctgaacg acacccgcta cgtgaaccgc
     5041 ttcctgtgcc agttcgtggc cgaccgcatg cgcctgaccg gcaagggcaa gaagcgcgtg
     5101 ttcgccagca acggccagat caccaacctg ctgcgcggct tctggggcct gcgcaaggtg
     5161 cgcgccgaga acgaccgcca ccacgccctg gacgccgtgg tggtggcctg cagcaccgtg
     5221 gccatgcagc agaagatcac ccgcttcgtg cgctacaagg agatgaacgc cttcgacggt
     5281 aaaaccatcg acaaggagac cggcgaggtg ctgcaccaga agacccactt cccccagccc
     5341 tgggagttct tcgcccagga ggtgatgatc cgcgtgttcg gcaagcccga cggcaagccc
     5401 gagttcgagg aggccgacac ccccgagaag ctgcgcaccc tgctggccga gaagctgagc
     5461 agccgccctg aggccgtgca cgagtacgtg actcctctgt tcgtgagccg cgcccccaac
     5521 cgcaagatga gcggtcaggg tcacatggag accgtgaaga gcgccaagcg cctggacgag
     5581 ggcgtgagcg tgctgcgcgt gcccctgacc cagctgaagc tgaaggacct ggagaagatg
     5641 gtgaaccgcg agcgcgagcc caagctgtac gaggccctga aggcccgcct ggaggcccac
     5701 aaggacgacc ccgccaaggc cttcgccgag cccttctaca agtacgacaa ggccggcaac
     5761 cgcacccagc aggtgaaggc cgtgcgcgtg gagcaggtgc agaagaccgg cgtgtgggtg
     5821 cgcaaccaca acggcatcgc cgacaacgcc accatggtgc gcgtggacgt gttcgagaag
     5881 ggcgacaagt actacctggt gcccatctac agctggcagg tggccaaggg catcctgccc
     5941 gaccgcgccg tggtgcaggg caaggacgag gaggactggc agctgatcga cgacagcttc
     6001 aacttcaagt tcagcctgca ccccaacgac ctggtggagg tgatcaccaa gaaggcccgc
     6061 atgttcggct acttcgccag ctgccaccgc ggcaccggca acatcaacat ccgcatccac
     6121 gacctggacc acaagatcgg caagaacggc atcctggagg gcatcggcgt gaagaccgcc
     6181 ctgagcttcc agaagtacca gatcgacgag ctgggcaagg agatccgccc ctgccgcctg
     6241 aagaagcgcc ctcctgtgcg cggctctact agtggctctc ccaagaagaa gaggaaggta
     6301 ggatccggaa gtatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg
     6361 gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac
     6421 gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggtacct
     6481 tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac
     6541 catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc
     6601 acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc
     6661 gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga cggcaatatc
     6721 ctgggccata agctggaata caattttaac agccacaatg tttacatcac cgccgataaa
     6781 caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg
     6841 cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca
     6901 gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat
     6961 catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg
     7021 tacaaatctg gcggccgctc tggcatgcgt aaaggcgaag agctgttcac tggtgtcgtc
     7081 cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag
     7141 ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa
     7201 ctgccggtac cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct
     7261 cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat
     7321 gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg
     7381 aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagaa
     7441 gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc
     7501 accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag
     7561 gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct
     7621 gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac
     7681 gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt
     7741 atggatgaac tgtacaaagg tagcatcgat ggtagcatgc gtaaaggcga agagctgttc
     7801 actggtgtcg tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc
     7861 gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt
     7921 actactggta aactgccggt accttggccg actctggtaa cgacgctgac ttatggtgtt
     7981 cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg
     8041 ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg
     8101 cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt
     8161 gactttaaag aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac
     8221 aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc
     8281 cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc
     8341 ggtgatggtc ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct
     8401 aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc
     8461 atcacgcatg gtatggatga actgtacaaa ctcgagtagg gtaatctaga taatcaacct
     8521 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     8581 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     8641 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     8701 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     8761 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     8821 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     8881 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     8941 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     9001 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     9061 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgagatcct ttaagaccaa
     9121 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     9181 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     9241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     9301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     9361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     9421 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     9481 ccggggcggc cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata
     9541 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     9601 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     9661 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     9721 ctttatatat cttgtggaaa ggacgaaaca ccgttagggt tagggttagg gttaggggtt
     9781 gtagctccct ttctcatttc ggaaacgaaa tgagaaccgt tgctacaata aggccgtctg
     9841 aaaagatgtg ccgcaacgct ctgcccctta aagcttctgc tttaaggggc atcgtttatt
     9901 tttttgaatt ctcgaccccc gggttaatta aggaaagggc tagatcattc ttgaagacga
     9961 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
    10021 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
    10081 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
    10141 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
    10201 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
    10261 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
    10321 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
    10381 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
    10441 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
    10501 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
    10561 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
    10621 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
    10681 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
    10741 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
    10801 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
    10861 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
    10921 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
    10981 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
    11041 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
    11101 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
    11161 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
    11221 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
    11281 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
    11341 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
    11401 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
    11461 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
    11521 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
    11581 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
    11641 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
    11701 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
    11761 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
    11821 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
    11881 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
    11941 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
    12001 cattaatgca gcaagctcat ggctgactaa ttttttttat ttatgcagag gccgaggccg
    12061 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
    12121 gcaaaaagct ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
    12181 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
    12241 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
    12301 atgattacga atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    12361 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
    12421 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
    12481 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
    12541 tgtactacaa acttagtagt
//
