LOCUS       sequence-492909-        8506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 and a sgRNA targeting the human RPA194 loci for 
            knock-in..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8506)
  AUTHORS   Ide S, Imai R, Ochi H, Maeshima K
  TITLE     Transcriptional suppression of ribosomal DNA with phase separation.
  JOURNAL   Sci Adv. 2020 Oct 14;6(42). pii: 6/42/eabb5953. doi: 
            10.1126/sciadv.abb5953. Print 2020 Oct.
  PUBMED    33055158
REFERENCE   2  (bases 1 to 8506)
  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: "Sci Adv."; 
            date: "2020-10-14"; volume: "6(42). pii: 6/42/eabb5953. doi"; pages:
            " 10.1126/sciadv.abb5953. Print 2020 Oct"
COMMENT     Sequence Label: pX330-RPA194
FEATURES             Location/Qualifiers
     source          1..8506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      47..502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(134..153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     344..365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(519..538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     638..660
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(698..716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        784..888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(889..957,958..1749)
                     /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(1107..1126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1920..2508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2409..2428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2570..2818
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2570..2590
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2741..2760
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2840..2915
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        3012..3297
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        3299..3574
                     /label=chicken beta-actin promoter
     intron          3575..3802
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      3814..3823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3823..3888
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3895..3915
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3940..8040
                     /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             3940..8040
                     /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             8041..8088
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8116..8133)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8122..8329
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8338..8478
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8338..8467
                     /label=AAV2 ITR
ORIGIN
        1 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
       61 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      121 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      181 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      241 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
      301 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      361 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
      421 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      481 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
      541 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      601 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      661 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
      721 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
      781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      901 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      961 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1021 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1081 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1141 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1201 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1261 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1321 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1381 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1441 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1501 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1561 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
     1621 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1681 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1741 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2041 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt
     2581 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     2641 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     2701 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     2761 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt
     2821 ggggtgtcgg aattcaaaag ttttagagct agaaatagca agttaaaata aggctagtcc
     2881 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag
     2941 caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct
     3001 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     3061 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
     3121 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     3181 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
     3241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     3301 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     3361 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggggsg
     3421 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
     3481 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     3541 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
     3601 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
     3661 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag
     3721 gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct
     3781 gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac
     3841 ggagactaca aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag
     3901 aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc
     3961 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     4021 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     4081 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     4141 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     4201 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     4261 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     4321 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     4381 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     4441 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     4501 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     4561 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     4621 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     4681 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     4741 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     4801 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     4861 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     4921 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     4981 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     5041 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     5101 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     5161 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     5221 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     5281 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     5341 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     5401 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     5461 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     5521 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     5581 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     5641 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     5701 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     5761 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     5821 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     5881 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     5941 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     6001 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     6061 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     6121 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     6181 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     6241 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     6301 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     6361 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     6421 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     6481 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     6541 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     6601 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     6661 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     6721 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     6781 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     6841 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     6901 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     6961 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     7021 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     7081 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     7141 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     7201 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     7261 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     7321 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     7381 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     7441 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     7501 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     7561 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     7621 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     7681 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     7741 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     7801 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     7861 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     7921 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     7981 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     8041 aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagta agaattccta
     8101 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     8161 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     8221 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     8281 aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag cggccgcagg
     8341 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     8401 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     8461 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttac
//
