LOCUS       sequence-262672-       11029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of FnCas9-GFP in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11029)
  AUTHORS   Acharya S, Mishra A, Paul D, Ansari AH, Azhar M, Kumar M, Rauthan R,
            Sharma N, Aich M, Sinha D, Sharma S, Jain S, Ray A, Jain S, 
            Ramalingam S, Maiti S, Chakraborty D
  TITLE     Francisella novicida Cas9 interrogates genomic DNA with very high 
            specificity and can be used for mammalian genome editing.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Oct 15;116(42):20959-20968. doi: 
            10.1073/pnas.1818461116. Epub 2019 Sep 30.
  PUBMED    31570623
REFERENCE   2  (bases 1 to 11029)
  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: 
            "10.1073/pnas.1818461116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-10-15- 15"; volume: "116"; issue: "42"; pages: 
            "20959-20968"
COMMENT     Sequence Label: pET-His6-FnCas9GFP
FEATURES             Location/Qualifiers
     source          1..11029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(549..1364)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(549..1364)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1272..1291
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1356..1375
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1486..2074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1975..1994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2228..2245
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2260..2399
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2325..2344)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2485..2507
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2501..2692)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3501..4583)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4545..4564
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4584..4661)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4867..4886
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4970..4989
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4970..4988
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4989..5013
                     /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)."
     RBS             5028..5050
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5069..5086
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(5096..5098,5099..5809)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5138..5159)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     5746..5767
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5819..5839
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5849..5875
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5879..5899
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5909..10792
                     /codon_start=1
                     /product="Cas9 endonuclease from the Francisella novicida 
                     Type II CRISPR/Cas system (Hirano et al., 2016)"
                     /label=FnCas9
                     /translation="NFKILPIAIDLGVKNTGVFSAFYQKGTSLERLDNKNGKVYELSKD
                     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             10862..10879
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(10932..10950)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      10946..10993
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaacta gtaacgttta caatttcagg tggcactttt cggggaaatg
      481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      541 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
      601 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
      661 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
      721 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
      781 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
      841 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
      901 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
      961 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatgtt ttcacctgaa
     1021 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     1081 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     1141 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     1201 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     1261 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     1321 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     1381 ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc
     1441 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1501 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1561 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1621 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1681 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1741 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1801 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1861 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1921 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1981 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2041 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2101 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2161 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2221 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2281 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     2341 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     2401 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2461 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2521 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     2581 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     2641 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     2701 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     2761 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     2821 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     2881 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     2941 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     3001 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     3061 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     3121 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     3181 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     3241 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     3301 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     3361 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     3421 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     3481 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3541 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     3601 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     3661 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     3721 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     3781 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     3841 tgatcgttgg caaccagcat cgcagtggga acgataccct cattcagcat ttgcatggtt
     3901 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     3961 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4021 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4081 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     4141 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     4201 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     4261 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     4321 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     4381 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     4441 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     4501 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     4561 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     4621 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     4681 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     4741 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     4801 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     4861 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     4921 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     4981 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     5041 aagaaggaga tataccatgg gttcttctca ccatcaccat caccatggtt cttctgtgag
     5101 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5161 aaacggccac aagttcagcg tgcgcggcga gggcgagggc gatgccacca acggcaagct
     5221 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5281 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     5341 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct ccttcaagga
     5401 cgacggcacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5461 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5521 gtacaacttc aacagccaca acgtctatat cacggccgac aagcagaaga acggcatcaa
     5581 ggcgaacttc aagatccgcc acaacgtcga ggacggcagc gtgcagctcg ccgaccacta
     5641 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5701 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5761 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg ggatcgagga
     5821 aaacctgtac ttccaatcca atgcaatgta cccatacgat gttccagatt acgcttcgcc
     5881 gaagaaaaag cgcaaggtcg aagcgtccaa ttttaagatc ctgcctatcg caatcgacct
     5941 gggcgtcaag aatactggcg tgtttagtgc tttttatcag aaggggacct cactggagag
     6001 actggacaat aagaacggaa aagtgtatga actgtccaag gattcttaca ctctgctgat
     6061 gaacaatagg accgcacgga gacaccagag gcgaggaatt gacaggaaac agctggtgaa
     6121 gcgcctgttc aaactgatct ggacagagca gctgaacctg gaatgggata aggacactca
     6181 gcaggccatc agcttcctgt ttaatcgacg gggattctct tttattactg acggctatag
     6241 tcctgagtac ctgaacatcg tgccagaaca ggtcaaggca atcctgatgg acattttcga
     6301 cgattataat ggcgaggacg atctggattc ctacctgaaa ctggccacag agcaagagag
     6361 taagatcagc gaaatctaca acaagctgat gcagaagatc ctggagttca agctgatgaa
     6421 actgtgcacc gacatcaagg acgataaagt gagtaccaag acactgaaag agatcacaag
     6481 ctacgagttc gaactgctgg ccgattatct ggctaactac agcgaatccc tgaagaccca
     6541 gaaattttcc tacacagaca agcagggcaa tctgaaagag ctgtcttact accaccatga
     6601 taagtacaac atccaggagt tcctgaagag acacgccacc atcaatgaca ggattctgga
     6661 tacactgctg actgacgatc tggacatctg gaacttcaac ttcgagaagt tcgatttcga
     6721 caagaacgag gaaaaactgc agaatcagga agataaggac cacattcagg ctcatctgca
     6781 ccatttcgtg tttgcagtca ataagatcaa aagcgagatg gcatccggcg ggcgccatcg
     6841 aagccagtac ttccaggaaa tcaccaacgt gctggacgag aacaatcacc aggaaggcta
     6901 cctgaaaaac ttctgtgaga atctgcataa caagaagtac agcaatctgt ccgtgaagaa
     6961 tctggtcaac ctgattggaa atctgtccaa cctggaactg aagcccctgc gcaaatactt
     7021 caacgacaag atccacgcta aagcagacca ttgggatgag cagaagttta ctgaaaccta
     7081 ttgccactgg attctgggcg agtggcgggt gggggtcaag gatcaggaca agaaagacgg
     7141 cgcaaagtat tcttacaagg acctgtgtaa cgagctgaag cagaaagtga ctaaggccgg
     7201 gctggtggac ttcctgctgg agctggaccc ctgccgaacc attccacctt acctggacaa
     7261 caataacaga aagccaccca aatgtcagag cctgatcctg aatcccaagt ttctggataa
     7321 tcagtatcct aactggcagc agtacctgca ggagctgaag aaactgcagt caatccagaa
     7381 ctacctggac agcttcgaaa ccgatctgaa ggtgctgaaa agctccaagg accagcctta
     7441 cttcgtcgag tacaagtcta gtaaccagca gatcgcttcc ggccagcggg attacaagga
     7501 tctggacgca agaatcctgc agttcatttt tgacagggtg aaggcctctg atgagctgct
     7561 gctgaacgaa atctatttcc aggcaaagaa actgaagcag aaagcctcaa gcgagctgga
     7621 aaagctggag tcctctaaga aactggacga agtgatcgct aactctcagc tgagtcagat
     7681 tctgaagtct cagcacacaa atggaatctt cgagcagggc acttttctgc atctggtgtg
     7741 caaatactat aagcagcgac agagagccag ggacagccgc ctgtacatca tgcctgaata
     7801 tcgatacgat aagaaactgc acaagtacaa caacaccggc cgctttgacg atgacaacca
     7861 gctgctgaca tattgtaatc ataagccccg gcagaaaaga taccagctgc tgaacgacct
     7921 ggcaggagtg ctgcaggtct ctcctaattt tctgaaggat aaaatcgggt ccgatgacga
     7981 tctgttcatt tctaagtggc tggtggagca catccggggc tttaagaagg cctgcgaaga
     8041 cagcctgaaa atccagaagg ataacagggg actgctgaat cataagatca acattgcacg
     8101 caataccaag ggcaaatgcg agaaagaaat cttcaacctg atctgtaaga ttgaggggag
     8161 cgaagacaag aaagggaatt ataagcacgg actggcctac gagctgggag tgctgctgtt
     8221 cggagagcca aacgaggcca gcaagcccga atttgatagg aaaatcaaga aattcaattc
     8281 aatctacagc tttgcccaga tccagcagat tgcctttgct gagaggaagg ggaatgcaaa
     8341 cacatgcgcc gtgtgtagtg cagacaacgc ccatcgcatg cagcagatca aaattactga
     8401 gccagtcgaa gacaataagg ataaaatcat tctgtcagca aaggcacagc gactgcctgc
     8461 aatcccaacc cgaattgtgg atggagctgt caagaaaatg gctacaattc tggcaaagaa
     8521 tatcgtggac gataattggc agaacattaa gcaggtcctg agcgcaaaac accagctgca
     8581 tatcccaatc attaccgagt ccaacgcctt cgagtttgaa cccgctctgg cagacgtgaa
     8641 gggcaaatct ctgaaggata gaaggaagaa agccctggag cgaattagtc ccgaaaacat
     8701 cttcaaggat aagaacaaca gaatcaagga gtttgctaag gggatttccg cctactctgg
     8761 agctaacctg acagatgggg acttcgatgg agcaaaggag gaactggatc acatcattcc
     8821 tcgcagccat aagaaatatg gcactctgaa cgacgaggct aatctgattt gcgtgacccg
     8881 gggcgataat aagaacaaag ggaaccggat cttctgtctg agagacctgg ccgataatta
     8941 caagctgaaa cagtttgaga ccacagacga tctggagatc gaaaagaaaa ttgccgacac
     9001 catctgggat gctaataaga aggacttcaa gttcggaaac tatcggagct tcatcaatct
     9061 gacacctcag gagcagaaag cattcagaca cgccctgttt ctggctgatg aaaacccaat
     9121 caagcaggca gtgatcagag ccattaataa ccgcaaccga accttcgtga atggcacaca
     9181 gaggtatttt gctgaggtcc tggcaaataa catctacctg cgcgccaaga aagaaaatct
     9241 gaacactgac aagatcagct tcgattactt tggaatccct accattggaa acggccgagg
     9301 gatcgctgag attcggcagc tgtatgaaaa ggtggacagt gatatccagg cctacgctaa
     9361 aggcgacaag ccacaggcct cttatagtca cctgattgat gctatgctgg cattctgcat
     9421 cgccgctgac gagcatcgga acgatggatc tattggcctg gaaatcgaca aaaactatag
     9481 tctgtaccct ctggataaga atactggcga ggtgttcacc aaagacatct tttcacagat
     9541 caagattacc gacaacgagt tcagcgataa gaaactggtc agaaagaaag ctattgaagg
     9601 gtttaacaca cacagacaga tgactaggga tggaatctat gcagagaatt acctgcctat
     9661 cctgattcat aaggagctga acgaagtgag gaaggggtac acatggaaaa attccgagga
     9721 aatcaaaatt ttcaagggaa agaaatacga catccagcag ctgaataacc tggtgtattg
     9781 tctgaagttt gtggacaaac caatcagtat tgatatccag atttcaaccc tggaggaact
     9841 gagaaacatc ctgactacca ataacattgc agccactgcc gagtactatt acattaatct
     9901 gaaaacccag aagctgcacg agtattacat cgaaaattac aacacagccc tggggtataa
     9961 gaaatacagc aaggagatgg agttcctgag gtccctggct tataggtctg agcgcgtgaa
    10021 gatcaaaagt attgacgatg tcaagcaggt cctggacaag gattcaaact tcatcatcgg
    10081 aaagatcaca ctgcccttca agaaagagtg gcagcgactg taccgggaat ggcagaacac
    10141 aactatcaaa gacgattatg agtttctgaa gagcttcttt aatgtgaagt ccattactaa
    10201 actgcacaag aaagtccgga aagacttctc tctgcccatc agtacaaacg agggcaagtt
    10261 tctggtgaag agaaaaactt gggataataa cttcatctac cagattctga atgactcaga
    10321 tagcagggca gacgggacta aacccttcat tcctgccttt gatatcagca agaacgagat
    10381 tgtggaagcc atcattgaca gtttcacctc aaaaaacatc ttttggctgc caaagaatat
    10441 tgagctgcag aaggtggaca acaagaacat cttcgccatt gataccagca agtggtttga
    10501 ggtcgaaaca ccatccgacc tgcgcgatat cggcattgct accattcagt acaagatcga
    10561 caataactca cgccccaagg tgcgagtcaa actggattac gtgatcgacg atgacagcaa
    10621 gattaactat ttcatgaatc actcactgct gaagagccgg tatcccgaca aagtcctgga
    10681 gatcctgaag cagagcacaa tcattgagtt cgaaagttca gggtttaaca aaactattaa
    10741 ggagatgctg ggaatgaagc tggccggcat ctacaatgaa acctccaata actaataaca
    10801 ttggaagtgg ataacggatc cgaattcgag cgccgtcgac aagcttgcgg ccgcactcga
    10861 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
    10921 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
    10981 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggg
//
