LOCUS       sequence-466565-        9409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one CRISPR/Cas9 vector with high-fidelity eSpCas9 expression 
            in neurons. The plasmid lacks AAV2 ITR and f1 ori elements, enabling
            more efficient transfection and expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9409)
  AUTHORS   Zhao C, Cao Y, Ibrahim N, Wang Y, Martemyanov KA
  TITLE     Efficient in vivo labeling of endogenous proteins with SMART 
            delineates retina cellular and synaptic organization.
  JOURNAL   Nat Commun. 2025 Apr 22;16(1):3768. doi: 10.1038/s41467-025-58945-6.
  PUBMED    40263339
REFERENCE   2  (bases 1 to 9409)
  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: "Nat 
            Commun."; date: "2025-04-22"; volume: "16(1):3768. doi"; pages: " 
            10.1038/s41467-025-58945-6"
COMMENT     Sequence Label: pCAG-eCas9-T2A-EGFP-U6-gRNA-no ITR and f1 ori
FEATURES             Location/Qualifiers
     source          1..9409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        523..826
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        828..1103
                     /label=chicken beta-actin promoter
     intron          1104..2120
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2044..2066
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2128..2147
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2195..2204
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2204..2269
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2276..2296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2321..4858,4859..4861,4862..5323,5324..5326,5327..5494,
                     5495..5497,5498..6421)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6422..6469
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6485..6538
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6539..6541,6542..7252)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6539..6541,6542..7252)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6842..6861)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7189..7210
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7280..7297)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7286..7493
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7538..7556)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7624..7728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7729..7797,7798..8589)
                     /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(7947..7966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8760..9348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9249..9268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agtaacggcc gccagtgtgc tggaattcag gctagttatt aatagtaatc
      481 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      541 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      601 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      661 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      721 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      781 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      841 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      901 tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggggcg ggccccgggg
      961 ggggggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
     1021 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1081 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
     1141 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
     1201 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
     1261 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
     1321 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
     1381 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     1441 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     1501 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     1561 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     1621 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     1681 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
     1741 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     1801 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     1861 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     1921 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     1981 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     2041 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     2101 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     2161 ttggcaaaga attcctgaat tctgcagatc cggtgccacc atggactata aggaccacga
     2221 cggagactac aaggatcatg atattgatta caaagacgat gacgataaga tggccccaaa
     2281 gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg
     2341 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     2401 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     2461 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
     2521 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     2581 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     2641 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     2701 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     2761 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     2821 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     2881 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     2941 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     3001 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     3061 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     3121 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     3181 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     3241 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     3301 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     3361 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     3421 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     3481 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     3541 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     3601 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     3661 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     3721 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     3781 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     3841 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     3901 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     3961 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     4021 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     4081 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     4141 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     4201 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     4261 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     4321 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     4381 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     4441 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     4501 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     4561 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     4621 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     4681 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     4741 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     4801 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctggc
     4861 cgacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     4921 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     4981 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     5041 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     5101 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     5161 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     5221 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     5281 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctgcgctgg aaagcgagtt
     5341 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     5401 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     5461 cgagattacc ctggccaacg gcgagatccg gaaggcgcct ctgatcgaga caaacggcga
     5521 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     5581 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     5641 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     5701 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     5761 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     5821 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     5881 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     5941 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     6001 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     6061 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     6121 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     6181 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
     6241 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
     6301 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
     6361 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
     6421 caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg aattcggcag
     6481 tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccagt
     6541 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     6601 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     6661 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     6721 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     6781 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     6841 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     6901 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     6961 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     7021 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     7081 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     7141 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     7201 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca aggaattcta
     7261 actagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     7321 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     7381 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     7441 gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagcggccg
     7501 catgcctgac caatgcagag tcgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     7561 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     7621 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7681 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7741 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7801 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7861 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7921 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7981 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     8041 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     8101 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     8161 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     8221 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     8281 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     8341 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     8401 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
     8461 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8521 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8581 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8641 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8701 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8761 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8821 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8881 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8941 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     9001 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     9061 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     9121 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     9181 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     9241 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     9301 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     9361 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
