LOCUS       sequence-105980-        8443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian gRNA expression vector also expressing SID4X-dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8443)
  TITLE     Dong Kong CRISPR plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8443)
  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: 
            "Unpublished"
COMMENT     Sequence Label: px330-SID4X-dCas9
FEATURES             Location/Qualifiers
     source          1..8443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             412..477
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             484..504
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(964..987,988..990,991..3477,3478..3480,3481..5067)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(964..4362,4363..4365,4366..4962,4963,4964..4966,
                     4967..4968,4969..4971,4972..5064)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5073..5090
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5113..5130)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5119..5343
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5414..5430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5414..5430)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5427..5449)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5438..5454
                     /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)."
     promoter        complement(join(5462..5468,5469..5486,5487..5492))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5507..5528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5645..5662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5816..6404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5896..5915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6575..7366,7367..7435))
                     /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     7198..7217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7436..7540)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7619..7638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7809..8057
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7809..7829
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7980..7999
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8076..8151
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        join(8271..8443,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggactacaaa
      421 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagatg
      481 gcccccaaga agaagaggaa ggtgggcatt caccgcgggg tacctggagg ttctggatcc
      541 atgaacatcc agatgctgct ggaggccgct gactacctgg aacggagaga gcgcgaagcc
      601 gagcacggat atgcttcaat gctgcccgga agcggcatga atattcagat gctgctggag
      661 gctgctgatt acctggaaag gcgcgaacgg gaggccgaac atggctatgc ttccatgctg
      721 cctgggtctg gaatgaatat ccaaatgctg ctggaggcag ccgattacct ggaacggaga
      781 gaaagagaag ccgagcacgg atacgccagc atgctgccag gcagcgggat gaacatacaa
      841 atgctgctgg aggctgccga ttacctggag aggcgcgaga gagaagctga acatggctat
      901 gcctctatgc tgcccagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa
      961 agtgataaaa agtattctat tggtttagct atcggcacta attccgttgg atgggctgtc
     1021 ataaccgatg aatacaaagt accttcaaag aaatttaagg tgttggggaa cacagaccgt
     1081 cattcgatta aaaagaatct tatcggtgcc ctcctattcg atagtggcga aacggcagag
     1141 gcgactcgcc tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa ccgaatatgt
     1201 tacttacaag aaatttttag caatgagatg gccaaagttg acgattcttt ctttcaccgt
     1261 ttggaagagt ccttccttgt cgaagaggac aagaaacatg aacggcaccc catctttgga
     1321 aacatagtag atgaggtggc atatcatgaa aagtacccaa cgatttatca cctcagaaaa
     1381 aagctagttg actcaactga taaagcggac ctgaggttaa tctacttggc tcttgcccat
     1441 atgataaagt tccgtgggca ctttctcatt gagggtgatc taaatccgga caactcggat
     1501 gtcgacaaac tgttcatcca gttagtacaa acctataatc agttgtttga agagaaccct
     1561 ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg cccgcctctc taaatcccga
     1621 cggctagaaa acctgatcgc acaattaccc ggagagaaga aaaatgggtt gttcggtaac
     1681 cttatagcgc tctcactagg cctgacacca aattttaagt cgaacttcga cttagctgaa
     1741 gatgccaaat tgcagcttag taaggacacg tacgatgacg atctcgacaa tctactggca
     1801 caaattggag atcagtatgc ggacttattt ttggctgcca aaaaccttag cgatgcaatc
     1861 ctcctatctg acatactgag agttaatact gagattacca aggcgccgtt atccgcttca
     1921 atgatcaaaa ggtacgatga acatcaccaa gacttgacac ttctcaaggc cctagtccgt
     1981 cagcaactgc ctgagaaata taaggaaata ttctttgatc agtcgaaaaa cgggtacgca
     2041 ggttatattg acggcggagc gagtcaagag gaattctaca agtttatcaa acccatatta
     2101 gagaagatgg atgggacgga agagttgctt gtaaaactca atcgcgaaga tctactgcga
     2161 aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa tccacttagg cgaattgcat
     2221 gctatactta gaaggcagga ggatttttat ccgttcctca aagacaatcg tgaaaagatt
     2281 gagaaaatcc taacctttcg cataccttac tatgtgggac ccctggcccg agggaactct
     2341 cggttcgcat ggatgacaag aaagtccgaa gaaacgatta ctccatggaa ttttgaggaa
     2401 gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccaa ctttgacaag
     2461 aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta tttcacagtg
     2521 tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc cgcctttcta
     2581 agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg caaagtgaca
     2641 gttaagcaat tgaaagagga ctactttaag aaaattgaat gcttcgattc tgtcgagatc
     2701 tccggggtag aagatcgatt taatgcgtca cttggtacgt atcatgacct cctaaagata
     2761 attaaagata aggacttcct ggataacgaa gagaatgaag atatcttaga agatatagtg
     2821 ttgactctta ccctctttga agatcgggaa atgattgagg aaagactaaa aacatacgct
     2881 cacctgttcg acgataaggt tatgaaacag ttaaagaggc gtcgctatac gggctgggga
     2941 cgattgtcgc ggaaacttat caacgggata agagacaagc aaagtggtaa aactattctc
     3001 gattttctaa agagcgacgg cttcgccaat aggaacttta tgcagctgat ccatgatgac
     3061 tctttaacct tcaaagagga tatacaaaag gcacaggttt ccggacaagg ggactcattg
     3121 cacgaacata ttgcgaatct tgctggttcg ccagccatca aaaagggcat actccagaca
     3181 gtcaaagtag tggatgagct agttaaggtc atgggacgtc acaaaccgga aaacattgta
     3241 atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag tcgagagcgg
     3301 atgaagagaa tagaagaggg tattaaagaa ctgggcagcc agatcttaaa ggagcatcct
     3361 gtggaaaata cccaattgca gaacgagaaa ctttacctct attacctaca aaatggaagg
     3421 gacatgtatg ttgatcagga actggacata aaccgtttat ctgattacga cgtcgatgcc
     3481 attgtacccc aatccttttt gaaggacgat tcaatcgaca ataaagtgct tacacgctcg
     3541 gataagaacc gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa gaaaatgaag
     3601 aactattggc ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt cgataactta
     3661 actaaagctg agaggggtgg cttgtctgaa cttgacaagg ccggatttat taaacgtcag
     3721 ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga tactagattc ccgaatgaat
     3781 acgaaatacg acgagaacga taagctgatt cgggaagtca aagtaatcac tttaaagtca
     3841 aaattggtgt cggacttcag aaaggatttt caattctata aagttaggga gataaataac
     3901 taccaccatg cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact cattaagaaa
     3961 tacccgaagc tagaaagtga gtttgtgtat ggtgattaca aagtttatga cgtccgtaag
     4021 atgatcgcga aaagcgaaca ggagataggc aaggctacag ccaaatactt cttttattct
     4081 aacattatga atttctttaa gacggaaatc actctggcaa acggagagat acgcaaacga
     4141 cctttaattg aaaccaatgg ggagacaggt gaaatcgtat gggataaggg ccgggacttc
     4201 gcgacggtga gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa aactgaggtg
     4261 cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga taagctcatc
     4321 gctcgtaaaa aggactggga cccgaaaaag tacggtggct tcgatagccc tacagttgcc
     4381 tattctgtcc tagtagtggc aaaagttgag aagggaaaat ccaagaaact gaagtcagtc
     4441 aaagaattat tggggataac gattatggag cgctcgtctt ttgaaaagaa ccccatcgac
     4501 ttccttgagg cgaaaggtta caaggaagta aaaaaggatc tcataattaa actaccaaag
     4561 tatagtctgt ttgagttaga aaatggccga aaacggatgt tggctagcgc cggagagctt
     4621 caaaagggga acgaactcgc actaccgtct aaatacgtga atttcctgta tttagcgtcc
     4681 cattacgaga agttgaaagg ttcacctgaa gataacgaac agaagcaact ttttgttgag
     4741 cagcacaaac attatctcga cgaaatcata gagcaaattt cggaattcag taagagagtc
     4801 atcctagctg atgccaatct ggacaaagta ttaagcgcat acaacaagca cagggataaa
     4861 cccatacgtg agcaggcgga aaatattatc catttgttta ctcttaccaa cctcggcgct
     4921 ccagccgcat tcaagtattt tgacacaacg atagatcgca aacgatacac ttctaccaag
     4981 gaggtgctag acgcgacact gattcaccaa tccatcacgg gattatatga aactcggata
     5041 gatttgtcac agcttggggg tgactgaccg gtcatcatca ccatcaccat tgagtttaaa
     5101 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     5161 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     5221 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     5281 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     5341 tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct
     5401 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5461 acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga gtgagctaac
     5521 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5581 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5641 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5701 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5761 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5821 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5881 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5941 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6001 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6061 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6121 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6181 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6241 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6301 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6361 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6421 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6481 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6541 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6601 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6661 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6721 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6781 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6841 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6901 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6961 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7021 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7081 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7141 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7201 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7261 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7321 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7381 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7441 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7501 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7561 cacctgacgt cgacggatcg ggagatcgat ctcccgatcc cctagggtcg actctcagta
     7621 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7681 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     7741 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     7801 atacgcgtga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     7861 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     7921 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     7981 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     8041 ggaaaggacg aaacaccggg tcttcgagaa gacctgtttt agagctagaa atagcaagtt
     8101 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
     8161 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgac tagttattaa
     8221 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     8281 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     8341 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggac
     8401 tatttacggt aaactgccca cttggcagta catcaagtgt atc
//
