LOCUS       sequence-430983-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pX330 based plasmid for expression of Cas9 and the 
            CGCTATCAAGATTTATACCT sgRNA to target the SHLD3 locus...
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  AUTHORS   Heyza JR, Mikhova M, Bahl A, Broadbent DG, Schmidt JC
  TITLE     Systematic analysis of the molecular and biophysical properties of 
            key DNA damage response factors.
  JOURNAL   Elife. 2023 Jun 21;12:e87086. doi: 10.7554/eLife.87086.
  PUBMED    37341699
REFERENCE   2  (bases 1 to 8508)
  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: "Elife."; 
            date: "2023-06-21"; volume: "12:e87086. doi"; pages: " 
            10.7554/eLife.87086"
COMMENT     Sequence Label: SHLD3 KO sgRNA #1
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..254
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6..26
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     177..196
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        276..351
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        448..733
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        735..1012
                     /label=chicken beta-actin promoter
     intron          1013..1240
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1252..1261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1261..1326
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1333..1353
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1378..5478
                     /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             1378..5478
                     /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             5479..5526
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5554..5571)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5560..5767
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5776..5916
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5776..5905
                     /label=AAV2 ITR
     rep_origin      5991..6446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6078..6097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6288..6309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6463..6482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6582..6604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6642..6660)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6728..6832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6833..6901,6902..7693)
                     /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(7051..7070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7864..8452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8353..8372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
       61 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      241 aaggacgaaa caccgcgcta tcaagattta tacctgtttt agagctagaa atagcaagtt
      301 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
      361 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
      421 ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
      481 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
      541 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      601 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
      661 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      721 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      781 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
      841 gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc
      901 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      961 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     1021 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     1081 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     1141 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
     1201 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
     1261 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
     1321 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
     1381 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
     1441 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     1501 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     1561 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     1621 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     1681 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     1741 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     1801 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     1861 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     1921 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     1981 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     2041 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     2101 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     2161 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     2221 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     2281 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     2341 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     2401 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     2461 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     2521 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     2581 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     2641 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     2701 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     2761 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     2821 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     2881 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     2941 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     3001 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     3061 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     3121 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     3181 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     3241 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     3301 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     3361 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     3421 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     3481 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     3541 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     3601 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     3661 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     3721 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     3781 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     3841 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     3901 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     3961 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     4021 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     4081 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     4141 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     4201 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     4261 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     4321 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     4381 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     4441 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     4501 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     4561 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     4621 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     4681 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     4741 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     4801 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     4861 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     4921 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     4981 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     5041 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     5101 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     5161 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     5221 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     5281 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     5341 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     5401 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     5461 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     5521 aaaaagtaag aattcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     5581 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     5641 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     5701 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg
     5761 ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     5821 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     5881 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     5941 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     6001 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6061 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6121 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6181 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     6241 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6301 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     6361 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6421 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     6481 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6541 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6601 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     6661 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     6721 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6781 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6841 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6901 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6961 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7021 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7081 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7141 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7201 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7261 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7321 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7381 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7441 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7501 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat
     7561 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7621 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7681 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7741 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7801 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7861 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7921 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7981 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8041 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8101 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8161 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8221 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     8281 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8341 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8401 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8461 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctca
//
