LOCUS       sequence-405308-       10046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of dead FnCas9 with T2A-EGFP and 
            cloning backbone for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10046)
  AUTHORS   Acharya S, Ansari AH, Kumar Das P, Hirano S, Aich M, Rauthan R, 
            Mahato S, Maddileti S, Sarkar S, Kumar M, Phutela R, Gulati S, 
            Rahman A, Goel A, Afzal C, Paul D, Agrawal T, Pulimamidi VK, Jalali 
            S, Nishimasu H, Mariappan I, Nureki O, Maiti S, Chakraborty D
  TITLE     PAM-flexible Engineered FnCas9 variants for robust and ultra-precise
            genome editing and diagnostics.
  JOURNAL   Nat Commun. 2024 Jun 28;15(1):5471. doi: 10.1038/s41467-024-49233-w.
  PUBMED    38942756
REFERENCE   2  (bases 1 to 10046)
  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: "Nat 
            Commun."; date: "2024-06-28"; volume: "15(1):5471. doi"; pages: " 
            10.1038/s41467-024-49233-w"
COMMENT     Sequence Label: PX458-3xHA-dFnCas9
FEATURES             Location/Qualifiers
     source          1..10046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        268..340
                     /label=Fn gRNA scaffold
                     /note="guide RNA scaffold for the Francisella novicida 
                     CRISPR/Cas9 system (Hirano et al., 2016)"
     enhancer        437..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          959..1186
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1198..1207
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1213..1293
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             1300..1320
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1345..6228
                     /codon_start=1
                     /product="Cas9 endonuclease from the Francisella novicida 
                     Type II CRISPR/Cas system (Hirano et al., 2016)"
                     /label=FnCas9
                     /translation="NFKILPIAIALGVKNTGVFSAFYQKGTSLERLDNKNGKVYELSKD
                     SYTLLMNNRTARRHQRRGIDRKQLVKRLFKLIWTEQLNLEWDKDTQQAISFLFNRRGFS
                     FITDGYSPEYLNIVPEQVKAILMDIFDDYNGEDDLDSYLKLATEQESKISEIYNKLMQK
                     ILEFKLMKLCTDIKDDKVSTKTLKEITSYEFELLADYLANYSESLKTQKFSYTDKQGNL
                     KELSYYHHDKYNIQEFLKRHATINDRILDTLLTDDLDIWNFNFEKFDFDKNEEKLQNQE
                     DKDHIQAHLHHFVFAVNKIKSEMASGGRHRSQYFQEITNVLDENNHQEGYLKNFCENLH
                     NKKYSNLSVKNLVNLIGNLSNLELKPLRKYFNDKIHAKADHWDEQKFTETYCHWILGEW
                     RVGVKDQDKKDGAKYSYKDLCNELKQKVTKAGLVDFLLELDPCRTIPPYLDNNNRKPPK
                     CQSLILNPKFLDNQYPNWQQYLQELKKLQSIQNYLDSFETDLKVLKSSKDQPYFVEYKS
                     SNQQIASGQRDYKDLDARILQFIFDRVKASDELLLNEIYFQAKKLKQKASSELEKLESS
                     KKLDEVIANSQLSQILKSQHTNGIFEQGTFLHLVCKYYKQRQRARDSRLYIMPEYRYDK
                     KLHKYNNTGRFDDDNQLLTYCNHKPRQKRYQLLNDLAGVLQVSPNFLKDKIGSDDDLFI
                     SKWLVEHIRGFKKACEDSLKIQKDNRGLLNHKINIARNTKGKCEKEIFNLICKIEGSED
                     KKGNYKHGMAYELGVLLFGEPNEASKPEFDRKIKKFNSIYSFAQIQQIAFAERKGNANT
                     CAVCSADNAHRMQQIKITEPVEDNKDKIILSAKAQRLPAIPTRIVDGAVKKMATILAKN
                     IVDDNWQNIKQVLSAKHQLHIPIITESNAFEFEPALADVKGKSLKDRRKKALERISPEN
                     IFKDKNNRIKEFAKGISAYSGANLTDGDFDGAKEELDAIIPRSHKKYGTLNDEANLICV
                     TRGDNKNKGNRIFCLRDLADNYKLKQFETTDDLEIEKKIADTIWDANKKDFKFGNYRSF
                     INLTPQEQKAFRHALFLADENPIKQAVIRAINNRNRTFVNGTQRYFAEVLANNIYLRAK
                     KENLNTDKISFDYFGIPTIGNGRGIAEIRQLYEKVDSDIQAYAKGDKPQASYSHLIDAM
                     LAFCIAADEHRNDGSIGLEIDKNYSLYPLDKNTGEVFTKDIFSQIKITDNEFSDKKLVR
                     KKAIEGFNTHRQMTRDGIYAENYLPILIHKELNEVRKGYTWKNSEEIKIFKGKKYDIQQ
                     LNNLVYCLKFVDKPISIDIQISTLEELRNILTTNNIAATAEYYYINLKTQKLHEYYIEN
                     YNTALGYKKYSKEMEFLRSLAYRSERVKIKSIDDVKQVLDKDSNFIIGKITLPFKKEWQ
                     RLYREWQNTTIKDDYEFLKSFFNVKSITKLHKKVRKDFSLPISTNEGKFLVKRKTWDNN
                     FIYQILNDSDSRADGTKPFIPAFDISKNEIVEAIIDSFTSKNIFWLPKNIELQKVDNKN
                     IFAIDTSKWFEVETPSDLRDIGIATIQYKIDNNSRPKVRVKLDYVIDDDSKINYFMNHS
                     LLKSRYPDKVLEILKQSTIIEFESSGFNKTIKEMLGMKLAGIYNETSNN"
     CDS             6229..6276
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6292..6345
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6346..6348,6349..7059)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6346..6348,6349..7059)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6388..6409)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6649..6668)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6996..7017
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7087..7104)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7093..7300
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7309..7449
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7309..7438
                     /label=AAV2 ITR
     rep_origin      7524..7979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7611..7630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7821..7842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7996..8015)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8115..8137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8175..8193)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8261..8365
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8366..8434,8435..9226)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8584..8603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9397..9985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9886..9905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt tcagttgcgc cgaaaggcgc tctgtaatca
      301 tttaaaagta ttttgaacgg acctctgttt gacacgtctg ttttttgttt tagagctaga
      361 aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa
      421 tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      481 acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat tgatgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca
      661 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
      781 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg aggggcgggg
      841 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      901 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      961 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     1021 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     1081 gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt
     1141 aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc
     1201 accatgggat cctacccata cgatgttcca gattacgctt acccatacga tgttccagat
     1261 tacgcttacc catacgatgt tccagattac gctatggccc caaagaagaa gcggaaggtc
     1321 ggtatccacg gagtcccagc agccaatttt aagatcctgc ctatcgcaat cgccctgggc
     1381 gtcaagaata ctggcgtgtt tagtgctttt tatcagaagg ggacctcact ggagagactg
     1441 gacaataaga acggaaaagt gtatgaactg tccaaggatt cttacactct gctgatgaac
     1501 aataggaccg cacggagaca ccagaggcga ggaattgaca ggaaacagct ggtgaagcgc
     1561 ctgttcaaac tgatctggac agagcagctg aacctggaat gggataagga cactcagcag
     1621 gccatcagct tcctgtttaa tcgacgggga ttctctttta ttactgacgg ctatagtcct
     1681 gagtacctga acatcgtgcc agaacaggtc aaggcaatcc tgatggacat tttcgacgat
     1741 tataatggcg aggacgatct ggattcctac ctgaaactgg ccacagagca agagagtaag
     1801 atcagcgaaa tctacaacaa gctgatgcag aagatcctgg agttcaagct gatgaaactg
     1861 tgcaccgaca tcaaggacga taaagtgagt accaagacac tgaaagagat cacaagctac
     1921 gagttcgaac tgctggccga ttatctggct aactacagcg aatccctgaa aacccagaaa
     1981 ttttcctaca cagacaagca gggcaatctg aaagagctgt cttactacca ccatgataag
     2041 tacaacatcc aggagttcct gaagagacac gccaccatca atgacaggat tctggataca
     2101 ctgctgactg acgatctgga catctggaac ttcaacttcg agaagttcga tttcgacaag
     2161 aacgaggaaa aactgcagaa tcaggaagat aaggaccaca ttcaggctca tctgcaccat
     2221 ttcgtgtttg cagtcaataa gatcaaaagc gagatggcat ccggcgggcg ccatcgaagc
     2281 cagtacttcc aggaaatcac caacgtgctg gacgagaaca atcaccagga aggctacctg
     2341 aaaaacttct gtgagaatct gcataacaag aagtacagca atctgtccgt gaagaatctg
     2401 gtcaacctga ttggaaatct gtccaacctg gaactgaagc ccctgcgcaa atacttcaac
     2461 gacaagatcc acgctaaagc agaccattgg gatgagcaga agtttactga aacctattgc
     2521 cactggattc tgggcgagtg gcgggtgggg gtcaaggatc aggacaagaa agacggcgca
     2581 aagtattctt acaaggacct gtgtaacgag ctgaagcaga aagtgactaa ggccgggctg
     2641 gtggacttcc tgctggagct ggacccctgc cgaaccattc caccttacct ggacaacaat
     2701 aacagaaagc cacccaaatg tcagagcctg atcctgaatc ccaagtttct ggataatcag
     2761 tatcctaact ggcagcagta cctgcaggag ctgaagaaac tgcagtcaat ccagaactac
     2821 ctggacagct tcgaaaccga tctgaaggtg ctgaaaagct ccaaggacca gccttacttc
     2881 gtcgagtaca agtctagtaa ccagcagatc gcttccggcc agcgggatta caaggatctg
     2941 gacgcaagaa tcctgcagtt catttttgac agggtgaagg cctctgatga gctgctgctg
     3001 aacgaaatct atttccaggc aaagaaactg aagcagaaag cctcaagcga gctggaaaag
     3061 ctggagtcct ctaagaaact ggacgaagtg atcgctaact ctcagctgag tcagattctg
     3121 aagtctcagc acacaaatgg aatcttcgag cagggcactt ttctgcatct ggtgtgcaaa
     3181 tactataagc agcgacagag agccagggac agccgcctgt acatcatgcc tgaatatcga
     3241 tacgataaga aactgcacaa gtacaacaac accggccgct ttgacgatga caaccagctg
     3301 ctgacatatt gtaatcataa gccccggcag aaaagatacc agctgctgaa cgacctggca
     3361 ggagtgctgc aggtctctcc taattttctg aaggataaaa tcgggtccga tgacgatctg
     3421 ttcatttcta agtggctggt ggagcacatc cggggcttta agaaggcctg cgaagatagc
     3481 ctgaaaatcc agaaggataa caggggactg ctgaatcata agatcaacat tgcacgcaat
     3541 accaagggca aatgcgagaa agaaatcttc aacctgatct gtaagattga ggggagcgaa
     3601 gataagaaag ggaattataa gcacggaatg gcctacgagc tgggagtgct gctgttcgga
     3661 gagccaaacg aggccagcaa gcccgaattt gataggaaaa tcaagaaatt caattcaatc
     3721 tacagctttg cccagatcca gcagattgcc tttgctgaga ggaaggggaa tgcaaacaca
     3781 tgcgccgtgt gtagtgcaga caacgcccat cgcatgcagc agatcaaaat tactgagcca
     3841 gtcgaagata ataaggataa aatcattctg tcagcaaagg cacagcgact gcctgcaatc
     3901 ccaacccgaa ttgtggatgg agctgtcaag aaaatggcta caattctggc aaagaatatc
     3961 gtggacgata attggcagaa cattaagcag gtcctgagcg caaaacacca gctgcatatc
     4021 ccaatcatta ccgagtccaa cgccttcgag tttgaacccg ctctggcaga cgtgaagggc
     4081 aaatctctga aggatagaag gaagaaagcc ctggagcgaa ttagtcccga aaacatcttc
     4141 aaggataaga acaacagaat caaggagttt gctaagggga tttccgccta ctctggagct
     4201 aacctgacag atggggactt cgatggagca aaggaggaac tggatgccat cattcctcgc
     4261 agccataaga aatatggcac tctgaacgac gaggctaatc tgatttgcgt gacccggggc
     4321 gataataaga acaaagggaa ccggatcttc tgtctgagag acctggccga taattacaag
     4381 ctgaaacagt ttgagaccac agacgatctg gagatcgaaa agaaaattgc cgacaccatc
     4441 tgggatgcta ataagaagga cttcaagttc ggaaactatc ggagcttcat caatctgaca
     4501 cctcaggagc agaaagcatt cagacacgcc ctgtttctgg ctgatgaaaa cccaatcaag
     4561 caggcagtga tcagagccat taataaccgc aaccgaacct tcgtgaatgg cacacagagg
     4621 tattttgctg aggtcctggc aaataacatc tacctgcgcg ccaagaaaga aaatctgaac
     4681 actgacaaga tcagcttcga ttactttgga atccctacca ttggaaacgg ccgagggatc
     4741 gctgagattc ggcagctgta tgaaaaggtg gacagtgata tccaggccta cgctaaaggc
     4801 gacaagccac aggcctctta tagtcacctg attgatgcta tgctggcatt ctgcatcgcc
     4861 gctgacgagc atcggaacga tggatctatt ggcctggaaa tcgacaaaaa ctatagtctg
     4921 taccctctgg ataagaatac tggcgaggtg ttcaccaaag acatcttttc acagatcaag
     4981 attaccgaca acgagttcag cgataagaaa ctggtcagaa agaaagctat tgaagggttt
     5041 aacacacaca gacagatgac tagggatgga atctatgcag agaattacct gcctatcctg
     5101 attcataagg agctgaacga agtgaggaag gggtacacat ggaaaaattc cgaggaaatc
     5161 aaaattttca agggaaagaa atacgacatc cagcagctga ataacctggt gtattgtctg
     5221 aagtttgtgg acaaaccaat cagtattgat atccagattt caaccctgga ggaactgaga
     5281 aacatcctga ctaccaataa cattgcagcc actgccgagt actattacat taatctgaaa
     5341 acccagaagc tgcacgagta ttacatcgaa aattacaaca cagccctggg gtataagaaa
     5401 tacagcaagg agatggagtt cctgaggtcc ctggcttata ggtctgagcg cgtgaagatc
     5461 aaaagtattg acgatgtcaa gcaggtcctg gacaaggatt caaacttcat catcggaaag
     5521 atcacactgc ccttcaagaa agagtggcag cgactgtacc gggaatggca gaacacaact
     5581 atcaaagacg attatgagtt tctgaagagc ttctttaatg tgaagtccat tactaaactg
     5641 cacaagaaag tccggaaaga cttctctctg cccatcagta caaacgaggg caagtttctg
     5701 gtgaagagaa aaacttggga taataacttc atctaccaga ttctgaatga ctcagatagc
     5761 agggcagacg ggactaaacc cttcattcct gcctttgata tcagcaagaa cgagattgtg
     5821 gaagccatca ttgacagttt cacctcaaaa aacatctttt ggctgccaaa gaatattgag
     5881 ctgcagaagg tggacaacaa gaacatcttc gccattgata ccagcaagtg gtttgaggtc
     5941 gaaacaccat ccgacctgcg cgatatcggc attgctacca ttcagtacaa gatcgacaat
     6001 aactcacgcc ccaaggtgcg agtcaaactg gattacgtga tcgacgatga cagcaagatt
     6061 aactatttca tgaatcactc actgctgaag agccggtatc ccgacaaagt cctggagatc
     6121 ctgaagcaga gcacaatcat tgagttcgaa agttcagggt ttaacaaaac tattaaggag
     6181 atgctgggaa tgaagctggc cggcatctac aatgaaacct ccaataacaa aaggccggcg
     6241 gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga
     6301 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag
     6361 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     6421 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     6481 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     6541 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     6601 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     6661 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     6721 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     6781 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     6841 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     6901 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     6961 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     7021 gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact agagctcgct
     7081 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     7141 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     7201 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     7261 agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag
     7321 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     7381 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     7441 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     7501 gcatacgtca aagcaaccat agtacgcgcc ttgtagcggc gcattaagcg cggcgggtgt
     7561 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     7621 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7681 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     7741 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7801 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     7861 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     7921 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     7981 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     8041 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     8101 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     8161 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     8221 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     8281 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     8341 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     8401 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8461 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     8521 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     8581 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     8641 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     8701 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8761 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     8821 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8881 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     8941 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     9001 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     9061 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     9121 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     9181 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     9241 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     9301 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     9361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     9421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     9481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     9541 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     9601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     9661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     9721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     9781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     9841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     9901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     9961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    10021 cttttgctgg ccttttgctc acatgt
//
