LOCUS       sequence-405304-       11029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of en1dFnCas9-GFP in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11029)
  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 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"; journalName: "Nat 
            Commun."; date: "2024-06-28"; volume: "15(1):5471. doi"; pages: " 
            10.1038/s41467-024-49233-w"
COMMENT     Sequence Label: pET-His6-en1dFnCas9GFP
FEATURES             Location/Qualifiers
     source          1..11029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(560..1375)
                     /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(560..1375)
                     /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     1283..1302
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1367..1386
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1497..2085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1986..2005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2239..2256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2271..2410
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2336..2355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2496..2518
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2512..2703)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3512..4594)
                     /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     4556..4575
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4595..4672)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4878..4897
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4981..5000
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4981..4999
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5000..5024
                     /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             5039..5061
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5080..5097
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(5107..5109,5110..5820)
                     /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(5149..5170)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     5757..5778
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5830..5850
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5860..5886
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5890..5910
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5920..10803
                     /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
                     IFKDKNNRIKEFAKGISAYSGANLTDGDFDGAKEELDAIIPRSHKKYGTLNDEANLICV
                     TRGDNKNKGNRIFCLRDLADNYKLKQFETTDDLEIEKKIADTIWDANKKDFKFGNYRSF
                     INLTPQEQKAFRHALFLADENPIKQAVIRAINNRNRTFVNGTQRYFAEVLANNIYLRAK
                     KENLNTDKISFDYFGIPTIGNGRGIAEIRQLYEKVDSDIQAYAKGDKPQASYSHLIDAM
                     LAFCIAADEHRNDGSIGLEIDKNYSLYPLDKNTGEVFTKDIFSQIKITDNEFSDKKLVR
                     KKAIEGFNTHRQMTRDGIYAENYLPILIHKELNEVRKGYTWKNSEEIKIFKGKKYDIQQ
                     LNNLVYCLKFVDKPISIDIQISTLEELRNILTTNNIAATAEYYYINLKTQKLHEYYIEN
                     YNTALGYKKYSKEMEFLRSLAYRSRRVKIKSIDDVKQVLDKDSNFIIGKITLPFKKEWQ
                     RLYREWQNTTIKDDYEFLKSFFNVKSITKLHKKVRKDFSLPISTNEGKFLVKRKTWDNN
                     FIYQILNDSDSRADGTKPFIPAFDISKNEIVEAIIDSFTSKNIFWLPKNIELQKVDNKN
                     IFAIDTSKWFEVETPSDLRDIGIATIQYKIDNNSRPKVRVKLDYVIDDDSKINYFMNHS
                     LLKSRYPDKVLEILKQSTIIEFESSGFNKTIKEMLGMKLAGIYNETSNN"
     CDS             10873..10890
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(10943..10961)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      10957..11004
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaact agtaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
      841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatgt
     1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     2341 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     2401 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2461 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2521 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     2581 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     2641 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     2701 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     2761 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     2821 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     2881 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     2941 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     3001 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     3061 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     3121 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     3181 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     3241 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     3301 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     3361 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     3421 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     3481 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3541 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3601 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     3661 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     3721 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     3781 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     3841 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgataccc tcattcagca
     3901 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     3961 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     4021 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     4081 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     4141 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     4201 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     4261 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     4321 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     4381 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     4441 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     4501 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     4561 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     4621 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     4681 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     4741 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     4801 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     4861 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     4921 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
     4981 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
     5041 tgtttaactt taagaaggag atataccatg ggttcttctc accatcacca tcaccatggt
     5101 tcttctgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     5161 gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc
     5221 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     5281 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     5341 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     5401 tccttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5461 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     5521 cacaagctgg agtacaactt caacagccac aacgtctata tcacggccga caagcagaag
     5581 aacggcatca aggcgaactt caagatccgc cacaacgtcg aggacggcag cgtgcagctc
     5641 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     5701 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     5761 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5821 gggatcgagg aaaacctgta cttccaatcc aatgcaatgt acccatacga tgttccagat
     5881 tacgcttcgc cgaagaaaaa gcgcaaggtc gaagcgtcca attttaagat cctgcctatc
     5941 gcaatcgccc tgggcgtcaa gaatactggc gtgtttagtg ctttttatca gaaggggacc
     6001 tcactggaga gactggacaa taagaacgga aaagtgtatg aactgtccaa ggattcttac
     6061 actctgctga tgaacaatag gaccgcacgg agacaccaga ggcgaggaat tgacaggaaa
     6121 cagctggtga agcgcctgtt caaactgatc tggacagagc agctgaacct ggaatgggat
     6181 aaggacactc agcaggccat cagcttcctg tttaatcgac ggggattctc ttttattact
     6241 gacggctata gtcctgagta cctgaacatc gtgccagaac aggtcaaggc aatcctgatg
     6301 gacattttcg acgattataa tggcgaggac gatctggatt cctacctgaa actggccaca
     6361 gagcaagaga gtaagatcag cgaaatctac aacaagctga tgcagaagat cctggagttc
     6421 aagctgatga aactgtgcac cgacatcaag gacgataaag tgagtaccaa gacactgaaa
     6481 gagatcacaa gctacgagtt cgaactgctg gccgattatc tggctaacta cagcgaatcc
     6541 ctgaagaccc agaaattttc ctacacagac aagcagggca atctgaaaga gctgtcttac
     6601 taccaccatg ataagtacaa catccaggag ttcctgaaga gacacgccac catcaatgac
     6661 aggattctgg atacactgct gactgacgat ctggacatct ggaacttcaa cttcgagaag
     6721 ttcgatttcg acaagaacga ggaaaaactg cagaatcagg aagataagga ccacattcag
     6781 gctcatctgc accatttcgt gtttgcagtc aataagatca aaagcgagat ggcatccggc
     6841 gggcgccatc gaagccagta cttccaggaa atcaccaacg tgctggacga gaacaatcac
     6901 caggaaggct acctgaaaaa cttctgtgag aatctgcata acaagaagta cagcaatctg
     6961 tccgtgaaga atctggtcaa cctgattgga aatctgtcca acctggaact gaagcccctg
     7021 cgcaaatact tcaacgacaa gatccacgct aaagcagacc attgggatga gcagaagttt
     7081 actgaaacct attgccactg gattctgggc gagtggcggg tgggggtcaa ggatcaggac
     7141 aagaaagacg gcgcaaagta ttcttacaag gacctgtgta acgagctgaa gcagaaagtg
     7201 actaaggccg ggctggtgga cttcctgctg gagctggacc cctgccgaac cattccacct
     7261 tacctggaca acaataacag aaagccaccc aaatgtcaga gcctgatcct gaatcccaag
     7321 tttctggata atcagtatcc taactggcag cagtacctgc aggagctgaa gaaactgcag
     7381 tcaatccaga actacctgga cagcttcgaa accgatctga aggtgctgaa aagctccaag
     7441 gaccagcctt acttcgtcga gtacaagtct agtaaccagc agatcgcttc cggccagcgg
     7501 gattacaagg atctggacgc aagaatcctg cagttcattt ttgacagggt gaaggcctct
     7561 gatgagctgc tgctgaacga aatctatttc caggcaaaga aactgaagca gaaagcctca
     7621 agcgagctgg aaaagctgga gtcctctaag aaactggacg aagtgatcgc taactctcag
     7681 ctgagtcaga ttctgaagtc tcagcacaca aatggaatct tcgagcaggg cacttttctg
     7741 catctggtgt gcaaatacta taagcagcga cagagagcca gggacagccg cctgtacatc
     7801 atgcctgaat atcgatacga taagaaactg cacaagtaca acaacaccgg ccgctttgac
     7861 gatgacaacc agctgctgac atattgtaat cataagcccc ggcagaaaag ataccagctg
     7921 ctgaacgacc tggcaggagt gctgcaggtc tctcctaatt ttctgaagga taaaatcggg
     7981 tccgatgacg atctgttcat ttctaagtgg ctggtggagc acatccgggg ctttaagaag
     8041 gcctgcgaag acagcctgaa aatccagaag gataacaggg gactgctgaa tcataagatc
     8101 aacattgcac gcaataccaa gggcaaatgc gagaaagaaa tcttcaacct gatctgtaag
     8161 attgagggga gcgaagacaa gaaagggaat tataagcacg gactggccta cgagctggga
     8221 gtgctgctgt tcggagagcc aaacgaggcc agcaagcccg aatttgatag gaaaatcaag
     8281 aaattcaatt caatctacag ctttgcccag atccagcaga ttgcctttgc tgagaggaag
     8341 gggaatgcaa acacatgcgc cgtgtgtagt gcagacaacg cccatcgcat gcagcagatc
     8401 aaaattactg agccagtcga agacaataag gataaaatca ttctgtcagc aaaggcacag
     8461 cgactgcctg caatcccaac ccgaattgtg gatggagctg tcaagaaaat ggctacaatt
     8521 ctggcaaaga atatcgtgga cgataattgg cagaacatta agcaggtcct gagcgcaaaa
     8581 caccagctgc atatcccaat cattaccgag tccaacgcct tcgagtttga acccgctctg
     8641 gcagacgtga agggcaaatc tctgaaggat agaaggaaga aagccctgga gcgaattagt
     8701 cccgaaaaca tcttcaagga taagaacaac agaatcaagg agtttgctaa ggggatttcc
     8761 gcctactctg gagctaacct gacagatggg gacttcgatg gagcaaagga ggaactggat
     8821 gccatcattc ctcgcagcca taagaaatat ggcactctga acgacgaggc taatctgatt
     8881 tgcgtgaccc ggggcgataa taagaacaaa gggaaccgga tcttctgtct gagagacctg
     8941 gccgataatt acaagctgaa acagtttgag accacagacg atctggagat cgaaaagaaa
     9001 attgccgaca ccatctggga tgctaataag aaggacttca agttcggaaa ctatcggagc
     9061 ttcatcaatc tgacacctca ggagcagaaa gcattcagac acgccctgtt tctggctgat
     9121 gaaaacccaa tcaagcaggc agtgatcaga gccattaata accgcaaccg aaccttcgtg
     9181 aatggcacac agaggtattt tgctgaggtc ctggcaaata acatctacct gcgcgccaag
     9241 aaagaaaatc tgaacactga caagatcagc ttcgattact ttggaatccc taccattgga
     9301 aacggccgag ggatcgctga gattcggcag ctgtatgaaa aggtggacag tgatatccag
     9361 gcctacgcta aaggcgacaa gccacaggcc tcttatagtc acctgattga tgctatgctg
     9421 gcattctgca tcgccgctga cgagcatcgg aacgatggat ctattggcct ggaaatcgac
     9481 aaaaactata gtctgtaccc tctggataag aatactggcg aggtgttcac caaagacatc
     9541 ttttcacaga tcaagattac cgacaacgag ttcagcgata agaaactggt cagaaagaaa
     9601 gctattgaag ggtttaacac acacagacag atgactaggg atggaatcta tgcagagaat
     9661 tacctgccta tcctgattca taaggagctg aacgaagtga ggaaggggta cacatggaaa
     9721 aattccgagg aaatcaaaat tttcaaggga aagaaatacg acatccagca gctgaataac
     9781 ctggtgtatt gtctgaagtt tgtggacaaa ccaatcagta ttgatatcca gatttcaacc
     9841 ctggaggaac tgagaaacat cctgactacc aataacattg cagccactgc cgagtactat
     9901 tacattaatc tgaaaaccca gaagctgcac gagtattaca tcgaaaatta caacacagcc
     9961 ctggggtata agaaatacag caaggagatg gagttcctga ggtccctggc ttataggtct
    10021 aggcgcgtga agatcaaaag tattgacgat gtcaagcagg tcctggacaa ggattcaaac
    10081 ttcatcatcg gaaagatcac actgcccttc aagaaagagt ggcagcgact gtaccgggaa
    10141 tggcagaaca caactatcaa agacgattat gagtttctga agagcttctt taatgtgaag
    10201 tccattacta aactgcacaa gaaagtccgg aaagacttct ctctgcccat cagtacaaac
    10261 gagggcaagt ttctggtgaa gagaaaaact tgggataata acttcatcta ccagattctg
    10321 aatgactcag atagcagggc agacgggact aaacccttca ttcctgcctt tgatatcagc
    10381 aagaacgaga ttgtggaagc catcattgac agtttcacct caaaaaacat cttttggctg
    10441 ccaaagaata ttgagctgca gaaggtggac aacaagaaca tcttcgccat tgataccagc
    10501 aagtggtttg aggtcgaaac accatccgac ctgcgcgata tcggcattgc taccattcag
    10561 tacaagatcg acaataactc acgccccaag gtgcgagtca aactggatta cgtgatcgac
    10621 gatgacagca agattaacta tttcatgaat cactcactgc tgaagagccg gtatcccgac
    10681 aaagtcctgg agatcctgaa gcagagcaca atcattgagt tcgaaagttc agggtttaac
    10741 aaaactatta aggagatgct gggaatgaag ctggccggca tctacaatga aacctccaat
    10801 aactaataac attggaagtg gataacggat ccgaattcga gcgccgtcga caagcttgcg
    10861 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
    10921 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
    10981 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggat
//
