LOCUS       sequence-170879-        9435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express gRNA and Cas9 in Leishmania with Hygromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9435)
  AUTHORS   Zhang WW, Lypaczewski P, Matlashewski G
  TITLE     Optimized CRISPR-Cas9 Genome Editing for Leishmania and Its Use To 
            Target a Multigene Family, Induce Chromosomal Translocation, and 
            Study DNA Break Repair Mechanisms.
  JOURNAL   mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 
            10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb.
  PUBMED    28124028
REFERENCE   2  (bases 1 to 9435)
  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: "mSphere."; 
            date: "2017-01-18"; volume: "2(1). pii: e00340-16. doi"; pages: " 
            10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb"
COMMENT     Sequence Label: pLdCH
FEATURES             Location/Qualifiers
     source          1..9435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..34
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(72..94)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     194..213
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_RNA        427..502
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           509..576
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     regulatory      744..753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             753..818
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             825..845
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             870..4970
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             870..4970
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4971..5018
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5046..5063)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(7382..7401)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7383..7401)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7488..7505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7659..8247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7739..7758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8418..9209,9210..9278))
                     /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     9041..9060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9279..9383)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
       61 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
      121 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
      181 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gtgagttatg
      241 aggtctgcga ttgacgtagg agttgcaagg ggagggggta tgaacggggt gggtagagct
      301 tttttaggtg gaagtagtga gagggtgggc ttgtgagaat ttgaggtgtg ttcgtgatgt
      361 gtggatctta tcgggggctc ggttgagttt ttggtttggt gattttgtgg gtcttcgaga
      421 agacctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      481 aaaagtggca ccgagtcggt gcttttttgg ccggcatggt cccagcctcc tcgctggcgc
      541 cggctgggca acatgcttcg gcatggcgaa tgggacggat ccccgggtac cgagctctct
      601 cttctccttc ccctctctct cctctctcct ctctccagag ctctctcttc tccttcccct
      661 ctctctcctc tctcctctct ccagagctcg aattcgcaag cttagcacct gcctgaaatc
      721 actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca
      781 aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga
      841 aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg
      901 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
      961 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     1021 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     1081 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     1141 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     1201 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     1261 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     1321 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     1381 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     1441 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     1501 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     1561 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     1621 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     1681 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     1741 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     1801 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     1861 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     1921 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     1981 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     2041 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     2101 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     2161 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     2221 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     2281 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     2341 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     2401 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     2461 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     2521 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     2581 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     2641 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     2701 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     2761 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     2821 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     2881 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     2941 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     3001 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     3061 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     3121 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     3181 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     3241 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     3301 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     3361 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     3421 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     3481 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     3541 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     3601 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     3661 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     3721 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     3781 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     3841 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     3901 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     3961 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     4021 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     4081 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     4141 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     4201 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     4261 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     4321 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     4381 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     4441 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     4501 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     4561 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     4621 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     4681 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     4741 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     4801 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     4861 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     4921 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg
     4981 cggccacgaa aaaggccggc caggcaaaaa agaaaaagta agaattccta gagctcgctg
     5041 atcagcctcg actgtgcctt ctagttgcca gccatctgtt ggatccgcgg ccccgcctcg
     5101 agcccgggcg ccgtcgaccg ctgcttgcgt tcgcatggac gccacgttcg acccgatcgg
     5161 gcagcaggcc gcggagacgc ggcagcgcaa acgtgcccgg cgggtggctc gttccggagg
     5221 aggcgttccg ccgtcgctac gtcagcatgc aagaggatca ttgctttgcg gttgaggtgg
     5281 cggagctcaa ctaccagcgt gccgtggaat acgttcatca agcgtcagcc gccggcgcgg
     5341 tcgcgacacc tgctcggcgg agcggccact gtgctgctct cttgacggcc ggtcgcgacg
     5401 cagggccgct ggtgggtctc cagcgggtac ggtgcgatca ggcagccatg acggctacgc
     5461 tgagcacgct tgcgaaggag ctggctgcac tgtcgccgta gcgcgatgcg agacgtatcg
     5521 agcgcatgcc agtggggtgc tgggcaacac ttggcggcgc gtggctcttt ctcactggtt
     5581 tgggcacgcc cacgctgtgg gtgtgtcggt gtctgccctg ccgctctctc tcctgatctc
     5641 tcgtcctggc aacaacaaca caccgcgaat gagttgccca caagccatct gcagtccgca
     5701 cgcgcgtccg ggccttgcgg tgtggatgcg agcatcgcgc attgccggcc accgccgcgt
     5761 gccaaaagca cacacatcgc tccccctcac gtggtgagcg tctgcgcgtg tgtgtctgtg
     5821 tgcataaatg cgtacgtgtg cgctcggtcc acggtcacca tcgcgccccg cggcatcggt
     5881 gctcgcacct ttccattctc tccgtacggc gttcgccact ctgctaactc ttctcccctt
     5941 tcctctctcc gcatctgcgc cttgtgtgcc gtgtgtgcgc cccacacgta ctctacacgc
     6001 cttctctgca tgcgctcctc gcctgcctga aacgatgacg tgcgccgcca ctcgctcacc
     6061 cgctcccatc cacatgcatt cctgcaccct acagcgactc taagccgtac attccatctc
     6121 ctcaacaccc actcgcttcc ttctgctctc acctattact tcgccagccc acatatctgc
     6181 tataacctgc cctcccccac ccgcttccca cacatccgcc accgctacga agcctcggta
     6241 ccgagctctc tcttctcctt cccctctctc tcctctctcc tctctccaga gctctctctt
     6301 ctccttcccc tctctctcct ctctcctctc tccagagctc gaattgatat gaaaaagcct
     6361 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     6421 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     6481 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     6541 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc
     6601 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     6661 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg
     6721 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     6781 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     6841 gttatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     6901 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     6961 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     7021 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     7081 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     7141 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     7201 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     7261 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     7321 ctcgccgata gtggaaacag acgccccagc actcgtccgg gatcatcaga tctgccggtc
     7381 tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc
     7441 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7501 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7561 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7621 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7681 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7741 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7801 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7861 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7921 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7981 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8041 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     8101 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8161 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8221 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8281 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8341 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8401 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8461 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8521 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8581 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8641 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8701 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8761 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8821 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8881 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8941 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9001 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9061 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9121 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9181 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9241 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9301 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9361 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     9421 accattatta tcatg
//
