LOCUS       sequence-405309-       11279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of FnCas9ABEmax8.17d base editor with 
            T2A-EGFP and cloning backbone for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11279)
  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 11279)
  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-FnCas9ABEmax8.17d
FEATURES             Location/Qualifiers
     source          1..11279
                     /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          974..1201
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1213..1222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1261..1281
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2470..7353
                     /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
                     KKGNYKHGLAYELGVLLFGEPNEASKPEFDRKIKKFNSIYSFAQIQQIAFAERKGNANT
                     CAVCSADNAHRMQQIKITEPVEDNKDKIILSAKAQRLPAIPTRIVDGAVKKMATILAKN
                     IVDDNWQNIKQVLSAKHQLHIPIITESNAFEFEPALADVKGKSLKDRRKKALERISPEN
                     IFKDKNNRIKEFAKGISAYSGANLTDGDFDGAKEELDHIIPRSHKKYGTLNDEANLICV
                     TRGDNKNKGNRIFCLRDLADNYKLKQFETTDDLEIEKKIADTIWDANKKDFKFGNYRSF
                     INLTPQEQKAFRHALFLADENPIKQAVIRAINNRNRTFVNGTQRYFAEVLANNIYLRAK
                     KENLNTDKISFDYFGIPTIGNGRGIAEIRQLYEKVDSDIQAYAKGDKPQASYSHLIDAM
                     LAFCIAADEHRNDGSIGLEIDKNYSLYPLDKNTGEVFTKDIFSQIKITDNEFSDKKLVR
                     KKAIEGFNTHRQMTRDGIYAENYLPILIHKELNEVRKGYTWKNSEEIKIFKGKKYDIQQ
                     LNNLVYCLKFVDKPISIDIQISTLEELRNILTTNNIAATAEYYYINLKTQKLHEYYIEN
                     YNTALGYKKYSKEMEFLRSLAYRSERVKIKSIDDVKQVLDKDSNFIIGKITLPFKKEWQ
                     RLYREWQNTTIKDDYEFLKSFFNVKSITKLHKKVRKDFSLPISTNEGKFLVKRKTWDNN
                     FIYQILNDSDSRADGTKPFIPAFDISKNEIVEAIIDSFTSKNIFWLPKNIELQKVDNKN
                     IFAIDTSKWFEVETPSDLRDIGIATIQYKIDNNSRPKVRVKLDYVIDDDSKINYFMNHS
                     LLKSRYPDKVLEILKQSTIIEFESSGFNKTIKEMLGMKLAGIYNETSNN"
     CDS             7396..7416
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7429..7509
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7525..7578
                     /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(7579..7581,7582..8292)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(7579..7581,7582..8292)
                     /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(7621..7642)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7882..7901)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8229..8250
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(8320..8337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8326..8533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8542..8682
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8542..8671
                     /label=AAV2 ITR
     rep_origin      8757..9212
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8844..8863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9054..9075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9229..9248)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9348..9370
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9408..9426)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9494..9598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9599..9667,9668..10459)
                     /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(9817..9836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10630..11218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11119..11138
                     /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 tgacgtcaat
      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 gggcggggcg
      841 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      901 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      961 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
     1021 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     1081 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
     1141 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
     1201 ggttggaccg gtgccaccat gggatccaaa cggacagccg acggaagcga gttcgagtca
     1261 ccaaagaaga agcggaaagt ctctgaagtc gagtttagcc acgagtattg gatgaggcac
     1321 gcactgaccc tggcaaagcg agcatgggat gaaagagaag tccccgtggg cgccgtgctg
     1381 gtgcacaaca atagagtgat cggagaggga tggaacaggc caatcggccg ccacgaccct
     1441 accgcacacg cagagatcat ggcactgagg cagggaggcc tggtcatgca gaattaccgc
     1501 ctgatcgatg ccaccctgta tgtgacactg gagccatgcg tgatgtgcgc aggagcaatg
     1561 atccacagca ggatcggaag agtggtgttc ggagcacggg acgccaagac cggcgcagca
     1621 ggctccctga tggatgtgct gcaccacccc ggcatgaacc accgggtgga gatcacagag
     1681 ggaatcctgg cagacgagtg cgccgccctg ctgagcgatt tctttagaat gcggagacag
     1741 gagatcaagg cccagaagaa ggcacagagc tccaccgact ctggaggatc tagcggagga
     1801 tcctctggaa gcgagacacc aggcacaagc gagtccgcca caccagagag ctccggcggc
     1861 tcctccggag gatcctctga ggtggagttt tcccacgagt actggatgag acatgccctg
     1921 accctggcca agagggcacg cgatgagagg gaggtgcctg tgggagccgt gctggtgctg
     1981 aacaatagag tgatcggcga gggctggaac agagccatcg gcctgcacga cccaacagcc
     2041 catgccgaaa ttatggccct gagacagggc ggcctggtca tgcagaacta cagactgatt
     2101 gacgccaccc tgtacagcac attcgagcct tgcgtgatgt gcgccggcgc catgatccac
     2161 tctaggatcg gccgcgtggt gtttggctgg aggaacgcaa aaaccggcgc cgcaggctcc
     2221 ctgatggacg tgctgcacta ccccggcatg aatcaccgcg tcgaaattac cgagggaatc
     2281 ctggcagatg aatgtgccgc cctgctgtgc tatttctttc ggatgcctag aagggtgttc
     2341 aatgctcaga agaaggccca gagctccacc gactccggag gatctagcgg aggctcctct
     2401 ggctctgaga cacctggcac aagcgagagc gcaacacctg aaagcagcgg gggcagcagc
     2461 ggggggtcaa attttaagat cctgcctatc gcaatcgccc tgggcgtcaa gaatactggc
     2521 gtgtttagtg ctttttatca gaaggggacc tcactggaga gactggacaa taagaacgga
     2581 aaagtgtatg aactgtccaa ggattcttac actctgctga tgaacaatag gaccgcacgg
     2641 agacaccaga ggcgaggaat tgacaggaaa cagctggtga agcgcctgtt caaactgatc
     2701 tggacagagc agctgaacct ggaatgggat aaggacactc agcaggccat cagcttcctg
     2761 tttaatcgac ggggattctc ttttattact gacggctata gtcctgagta cctgaacatc
     2821 gtgccagaac aggtcaaggc aatcctgatg gacattttcg acgattataa tggcgaggac
     2881 gatctggatt cctacctgaa actggccaca gagcaagaga gtaagatcag cgaaatctac
     2941 aacaagctga tgcagaagat cctggagttc aagctgatga aactgtgcac cgacatcaag
     3001 gacgataaag tgagtaccaa gacactgaaa gagatcacaa gctacgagtt cgaactgctg
     3061 gccgattatc tggctaacta cagcgaatcc ctgaaaaccc agaaattttc ctacacagac
     3121 aagcagggca atctgaaaga gctgtcttac taccaccatg ataagtacaa catccaggag
     3181 ttcctgaaga gacacgccac catcaatgac aggattctgg atacactgct gactgacgat
     3241 ctggacatct ggaacttcaa cttcgagaag ttcgatttcg acaagaacga ggaaaaactg
     3301 cagaatcagg aagataagga ccacattcag gctcatctgc accatttcgt gtttgcagtc
     3361 aataagatca aaagcgagat ggcatccggc gggcgccatc gaagccagta cttccaggaa
     3421 atcaccaacg tgctggacga gaacaatcac caggaaggct acctgaaaaa cttctgtgag
     3481 aatctgcata acaagaagta cagcaatctg tccgtgaaga atctggtcaa cctgattgga
     3541 aatctgtcca acctggaact gaagcccctg cgcaaatact tcaacgacaa gatccacgct
     3601 aaagcagacc attgggatga gcagaagttt actgaaacct attgccactg gattctgggc
     3661 gagtggcggg tgggggtcaa ggatcaggac aagaaagacg gcgcaaagta ttcttacaag
     3721 gacctgtgta acgagctgaa gcagaaagtg actaaggccg ggctggtgga cttcctgctg
     3781 gagctggacc cctgccgaac cattccacct tacctggaca acaataacag aaagccaccc
     3841 aaatgtcaga gcctgatcct gaatcccaag tttctggata atcagtatcc taactggcag
     3901 cagtacctgc aggagctgaa gaaactgcag tcaatccaga actacctgga cagcttcgaa
     3961 accgatctga aggtgctgaa aagctccaag gaccagcctt acttcgtcga gtacaagtct
     4021 agtaaccagc agatcgcttc cggccagcgg gattacaagg atctggacgc aagaatcctg
     4081 cagttcattt ttgacagggt gaaggcctct gatgagctgc tgctgaacga aatctatttc
     4141 caggcaaaga aactgaagca gaaagcctca agcgagctgg aaaagctgga gtcctctaag
     4201 aaactggacg aagtgatcgc taactctcag ctgagtcaga ttctgaagtc tcagcacaca
     4261 aatggaatct tcgagcaggg cacttttctg catctggtgt gcaaatacta taagcagcga
     4321 cagagagcca gggacagccg cctgtacatc atgcctgaat atcgatacga taagaaactg
     4381 cacaagtaca acaacaccgg ccgctttgac gatgacaacc agctgctgac atattgtaat
     4441 cataagcccc ggcagaaaag ataccagctg ctgaacgacc tggcaggagt gctgcaggtc
     4501 tctcctaatt ttctgaagga taaaatcggg tccgatgacg atctgttcat ttctaagtgg
     4561 ctggtggagc acatccgggg ctttaagaag gcctgcgaag atagcctgaa aatccagaag
     4621 gataacaggg gactgctgaa tcataagatc aacattgcac gcaataccaa gggcaaatgc
     4681 gagaaagaaa tcttcaacct gatctgtaag attgagggga gcgaagataa gaaagggaat
     4741 tataagcacg gactggccta cgagctggga gtgctgctgt tcggagagcc aaacgaggcc
     4801 agcaagcccg aatttgatag gaaaatcaag aaattcaatt caatctacag ctttgcccag
     4861 atccagcaga ttgcctttgc tgagaggaag gggaatgcaa acacatgcgc cgtgtgtagt
     4921 gcagacaacg cccatcgcat gcagcagatc aaaattactg agccagtcga agataataag
     4981 gataaaatca ttctgtcagc aaaggcacag cgactgcctg caatcccaac ccgaattgtg
     5041 gatggagctg tcaagaaaat ggctacaatt ctggcaaaga atatcgtgga cgataattgg
     5101 cagaacatta agcaggtcct gagcgcaaaa caccagctgc atatcccaat cattaccgag
     5161 tccaacgcct tcgagtttga acccgctctg gcagacgtga agggcaaatc tctgaaggat
     5221 agaaggaaga aagccctgga gcgaattagt cccgaaaaca tcttcaagga taagaacaac
     5281 agaatcaagg agtttgctaa ggggatttcc gcctactctg gagctaacct gacagatggg
     5341 gacttcgatg gagcaaagga ggaactggat cacatcattc ctcgcagcca taagaaatat
     5401 ggcactctga acgacgaggc taatctgatt tgcgtgaccc ggggcgataa taagaacaaa
     5461 gggaaccgga tcttctgtct gagagacctg gccgataatt acaagctgaa acagtttgag
     5521 accacagacg atctggagat cgaaaagaaa attgccgaca ccatctggga tgctaataag
     5581 aaggacttca agttcggaaa ctatcggagc ttcatcaatc tgacacctca ggagcagaaa
     5641 gcattcagac acgccctgtt tctggctgat gaaaacccaa tcaagcaggc agtgatcaga
     5701 gccattaata accgcaaccg aaccttcgtg aatggcacac agaggtattt tgctgaggtc
     5761 ctggcaaata acatctacct gcgcgccaag aaagaaaatc tgaacactga caagatcagc
     5821 ttcgattact ttggaatccc taccattgga aacggccgag ggatcgctga gattcggcag
     5881 ctgtatgaaa aggtggacag tgatatccag gcctacgcta aaggcgacaa gccacaggcc
     5941 tcttatagtc acctgattga tgctatgctg gcattctgca tcgccgctga cgagcatcgg
     6001 aacgatggat ctattggcct ggaaatcgac aaaaactata gtctgtaccc tctggataag
     6061 aatactggcg aggtgttcac caaagacatc ttttcacaga tcaagattac cgacaacgag
     6121 ttcagcgata agaaactggt cagaaagaaa gctattgaag ggtttaacac acacagacag
     6181 atgactaggg atggaatcta tgcagagaat tacctgccta tcctgattca taaggagctg
     6241 aacgaagtga ggaaggggta cacatggaaa aattccgagg aaatcaaaat tttcaaggga
     6301 aagaaatacg acatccagca gctgaataac ctggtgtatt gtctgaagtt tgtggacaaa
     6361 ccaatcagta ttgatatcca gatttcaacc ctggaggaac tgagaaacat cctgactacc
     6421 aataacattg cagccactgc cgagtactat tacattaatc tgaaaaccca gaagctgcac
     6481 gagtattaca tcgaaaatta caacacagcc ctggggtata agaaatacag caaggagatg
     6541 gagttcctga ggtccctggc ttataggtct gagcgcgtga agatcaaaag tattgacgat
     6601 gtcaagcagg tcctggacaa ggattcaaac ttcatcatcg gaaagatcac actgcccttc
     6661 aagaaagagt ggcagcgact gtaccgggaa tggcagaaca caactatcaa agacgattat
     6721 gagtttctga agagcttctt taatgtgaag tccattacta aactgcacaa gaaagtccgg
     6781 aaagacttct ctctgcccat cagtacaaac gagggcaagt ttctggtgaa gagaaaaact
     6841 tgggataata acttcatcta ccagattctg aatgactcag atagcagggc agacgggact
     6901 aaacccttca ttcctgcctt tgatatcagc aagaacgaga ttgtggaagc catcattgac
     6961 agtttcacct caaaaaacat cttttggctg ccaaagaata ttgagctgca gaaggtggac
     7021 aacaagaaca tcttcgccat tgataccagc aagtggtttg aggtcgaaac accatccgac
     7081 ctgcgcgata tcggcattgc taccattcag tacaagatcg acaataactc acgccccaag
     7141 gtgcgagtca aactggatta cgtgatcgac gatgacagca agattaacta tttcatgaat
     7201 cactcactgc tgaagagccg gtatcccgac aaagtcctgg agatcctgaa gcagagcaca
     7261 atcattgagt tcgaaagttc agggtttaac aaaactatta aggagatgct gggaatgaag
     7321 ctggccggca tctacaatga aacctccaat aactctggcg gctcaaaaag aaccgccgac
     7381 ggcagcgagt tcgagcccaa gaagaagagg aaagtcagcg gcggcagcta cccatacgat
     7441 gttccagatt acgcttaccc atacgatgtt ccagattacg cttacccata cgatgttcca
     7501 gattacgctg aattcggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc
     7561 gaggagaatc ctggcccagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     7621 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     7681 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     7741 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     7801 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     7861 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     7921 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     7981 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     8041 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     8101 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     8161 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     8221 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     8281 gagctgtaca aggagttcta actagagctc gctgatcagc ctcgactgtg ccttctagtt
     8341 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     8401 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     8461 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca
     8521 ggcatgctgg ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     8581 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     8641 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     8701 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     8761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     8821 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     8881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     8941 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     9001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     9061 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     9121 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     9181 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     9241 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     9301 ggcttgtctg ctcccagcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     9361 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     9421 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     9481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     9541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     9601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     9661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     9721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     9781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     9841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     9901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     9961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
    10021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
    10081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
    10141 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
    10201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
    10261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg
    10321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    10381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    10441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    10501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    10561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    10621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    10681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    10741 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    10801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    10861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    10921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    10981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    11041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    11101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    11161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    11221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
