LOCUS       Exported                9416 bp ds-DNA     circular SYN 13-JUN-2023
DEFINITION  All-in-one CRISPR/Cas9 vector with high-fidelity eSpCas9 expression 
            in human pluripotent stem cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-eCas9-GFP-U6-gRNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9416)
  TITLE     pCAG-SpCas9-U6-gRNA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 18, 2025 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9416
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     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"
     primer_bind     246..270
                     /label=F
     primer_bind     complement(250..274)
                     /label=Primer 1
     misc_feature    251..270
                     /label=gRNA
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        526..829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        831..1108
                     /label=chicken beta-actin promoter
     intron          1110..2127
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2051..2073
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2135..2154
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2202..2211
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2211..2276
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2283..2303
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2328..6428
                     /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"
     primer_bind     complement(5931..5951)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             6429..6476
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6492..6545
                     /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             6546..7259
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             6546..7259
                     /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(6588..6609)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6849..6868)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7196..7217
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7287..7304)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7293..7500
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    7503..7532
                     /label=Feature 21
     primer_bind     7503..7528
                     /label=insert_F
     primer_bind     complement(7507..7532)
                     /label=insert_R
     promoter        7631..7735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7736..8596
                     /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(7954..7973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8767..9355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9256..9275
                     /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 gtatggtgtg gagcccaacg gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagtaacg gccgccagtg tgctggaatt caggctagtt attaatagta
      481 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      541 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      601 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      661 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      721 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      781 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      841 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      901 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc
      961 aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc
     1021 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     1081 ctataaaaag cgaagcgcgc ggcgggcggg gagtcgctgc gacgctgcct tcgccccgtg
     1141 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     1201 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     1261 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
     1321 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
     1381 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1441 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc
     1501 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc
     1561 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg
     1621 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg
     1681 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg
     1741 gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat
     1801 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg
     1861 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc
     1921 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
     1981 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
     2041 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     2101 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     2161 catcattttg gcaaagaatt cctgaattct gcagatccgg tgccaccatg gactataagg
     2221 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg
     2281 ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
     2341 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     2401 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     2461 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     2521 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     2581 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     2641 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     2701 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     2761 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     2821 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     2881 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     2941 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     3001 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     3061 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     3121 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     3181 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     3241 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     3301 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     3361 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     3421 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     3481 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     3541 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     3601 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     3661 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     3721 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     3781 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     3841 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     3901 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     3961 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     4021 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     4081 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     4141 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     4201 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     4261 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     4321 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     4381 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     4441 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     4501 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     4561 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     4621 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     4681 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     4741 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     4801 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     4861 ttctggccga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     4921 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     4981 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     5041 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     5101 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     5161 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     5221 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     5281 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct gcgctggaaa
     5341 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     5401 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     5461 tcaagaccga gattaccctg gccaacggcg agatccggaa ggcgcctctg atcgagacaa
     5521 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     5581 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     5641 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     5701 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     5761 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     5821 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     5881 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     5941 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     6001 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     6061 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     6121 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     6181 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     6241 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     6301 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     6361 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     6421 gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat
     6481 tcggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     6541 gcccagtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     6601 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     6661 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     6721 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     6781 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     6841 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     6901 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     6961 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     7021 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     7081 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     7141 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     7201 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     7261 aattctaact agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     7321 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     7381 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     7441 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
     7501 gcggccgcat gcctgaccaa tgcagagtcg cgcgagacga aagggcctcg tgatacgcct
     7561 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     7621 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     7681 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     7741 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     7801 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     7861 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     7921 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     7981 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     8041 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     8101 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     8161 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     8221 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     8281 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     8341 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     8401 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg
     8461 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     8521 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     8581 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     8641 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     8701 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8761 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8821 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8881 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     8941 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     9001 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     9061 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     9121 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     9181 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     9241 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     9301 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     9361 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
