LOCUS       sequence-420349-        8956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9 and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8956)
  AUTHORS   Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T
  TITLE     Production of knockout mice by DNA microinjection of various 
            CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes.
  JOURNAL   BMC Biotechnol. 2015 May 22;15(1):33.
  PUBMED    25997509
REFERENCE   2  (bases 1 to 8956)
  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: "BMC 
            Biotechnol."; date: "2015-05-22"; volume: "15(1)"; pages: "33"
COMMENT     Sequence Label: pX330A_dCas9-1x6
FEATURES             Location/Qualifiers
     source          1..8956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..293
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        295..569
                     /label=chicken beta-actin promoter
     intron          570..797
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      809..818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             818..883
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             890..910
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             935..5035
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIYLSQLGGD"
     CDS             5036..5083
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5111..5128)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5117..5324
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5333..5473
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5333..5462
                     /label=AAV2 ITR
     rep_origin      5548..6003
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5635..5654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5845..5866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6020..6039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6139..6161
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6199..6217)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6285..6389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6390..6458,6459..7250)
                     /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(6608..6627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7421..8009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7910..7929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8071..8319
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8071..8091
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8242..8261
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8338..8413
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             complement(8499..8804)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     8654..8676
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8668..8685
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8669..8685
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8695..8714
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8695..8713
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8800..8816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8800..8816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8813..8835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8824..8840
                     /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(8848..8854,8855..8872,8873..8878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8893..8914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      121 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      181 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
      241 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
      301 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
      361 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg gggcgcgcgc
      421 caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag
      481 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
      541 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
      601 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
      661 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag
      721 ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg
      781 aaatcacttt ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga
      841 ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa
      901 gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctggc
      961 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     1021 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     1081 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     1141 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     1201 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     1261 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     1321 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     1381 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     1441 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     1501 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     1561 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     1621 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     1681 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     1741 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     1801 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     1861 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     1921 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     1981 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     2041 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     2101 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     2161 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     2221 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     2281 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     2341 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     2401 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     2461 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     2521 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     2581 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     2641 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     2701 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     2761 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     2821 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     2881 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     2941 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     3001 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     3061 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     3121 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     3181 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     3241 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     3301 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     3361 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     3421 caaccggctg tccgactacg atgtggacgc catcgtgcct cagagctttc tgaaggacga
     3481 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     3541 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     3601 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     3661 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     3721 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     3781 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     3841 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     3901 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     3961 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     4021 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     4081 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     4141 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     4201 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     4261 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     4321 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     4381 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     4441 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     4501 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     4561 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     4621 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     4681 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     4741 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     4801 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     4861 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     4921 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     4981 gagcatcacc ggcctgtacg agacacggat ctacctgtct cagctgggag gcgacaaaag
     5041 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aagtaagaat tcctagagct
     5101 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     5161 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     5221 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     5281 agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc
     5341 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     5401 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     5461 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5521 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     5581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     5641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5761 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     5881 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     5941 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     6001 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     6061 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     6121 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     6181 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     6241 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6361 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6421 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6481 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6541 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6601 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6661 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6721 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6781 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6841 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6901 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6961 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7021 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7081 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg
     7141 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7201 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7261 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7321 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7381 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7441 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7501 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7561 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7621 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7681 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7741 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7801 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7861 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7921 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7981 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8041 tggccttttg ctggcctttt gctcacatgt gagggcctat ttcccatgat tccttcatat
     8101 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
     8161 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     8221 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     8281 ttcttggctt tatatatctt gtggaaagga cgaaacaccg ggtcttcgag aagacctgtt
     8341 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     8401 accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc
     8461 gtttttagcg cgtgcgccaa ttctgcagac aaatggctct agagaccggc gccgctacag
     8521 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8581 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8641 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     8701 gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc
     8761 cccttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     8821 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     8881 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     8941 ccaccggtgg tctctg
//
