LOCUS       Exported                9413 bp ds-DNA     circular SYN 03-APR-2024
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 9413)
  TITLE     pCAG-SpCas9-U6-gRNA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 18, 2025 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9413
                     /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"
     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..1105
                     /label=chicken beta-actin promoter
     intron          1107..2124
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2048..2070
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2132..2151
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2199..2208
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2208..2273
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2280..2300
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2325..6425
                     /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     3961..3981
                     /label=Cas9_F1
     primer_bind     complement(4210..4230)
                     /label=Cas9_R2
     primer_bind     complement(4345..4365)
                     /label=Cas9_R1
     primer_bind     complement(5928..5948)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             6426..6473
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6489..6542
                     /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             6543..7256
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             6543..7256
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6585..6606)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6846..6865)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7193..7214
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7284..7301)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7290..7497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    7500..7529
                     /label=Feature 21
     primer_bind     7500..7525
                     /label=insert_F
     primer_bind     complement(7504..7529)
                     /label=insert_R
     promoter        7628..7732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7733..8593
                     /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(7951..7970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8764..9352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9253..9272
                     /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 gggggggggc gcgcgccagg
      961 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
     1021 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
     1081 taaaaagcga agcgcgcggc gggcggggag tcgctgcgac gctgccttcg ccccgtgccc
     1141 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     1201 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     1261 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
     1321 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
     1381 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     1441 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
     1501 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     1561 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
     1621 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     1681 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
     1741 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
     1801 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
     1861 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
     1921 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
     1981 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     2041 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     2101 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     2161 cattttggca aagaattcct gaattctgca gatccggtgc caccatggac tataaggacc
     2221 acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc
     2281 caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca
     2341 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     2401 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     2461 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     2521 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     2581 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     2641 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     2701 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     2761 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     2821 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     2881 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     2941 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     3001 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     3061 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     3121 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     3181 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     3241 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     3301 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     3361 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     3421 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     3481 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     3541 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     3601 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     3661 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     3721 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     3781 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     3841 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     3901 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     3961 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     4021 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     4081 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     4141 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     4201 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     4261 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     4321 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     4381 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     4441 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     4501 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     4561 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     4621 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     4681 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     4741 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     4801 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     4861 tggccgacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     4921 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     4981 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     5041 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     5101 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     5161 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     5221 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     5281 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctgcg ctggaaagcg
     5341 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     5401 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     5461 agaccgagat taccctggcc aacggcgaga tccggaaggc gcctctgatc gagacaaacg
     5521 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     5581 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     5641 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     5701 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     5761 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     5821 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     5881 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     5941 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     6001 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     6061 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     6121 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     6181 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     6241 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     6301 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     6361 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     6421 gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg
     6481 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     6541 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     6601 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     6661 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     6721 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     6781 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     6841 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     6901 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     6961 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     7021 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     7081 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     7141 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     7201 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggaat
     7261 tctaactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     7321 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     7381 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     7441 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
     7501 gccgcatgcc tgaccaatgc agagtcgcgc gagacgaaag ggcctcgtga tacgcctatt
     7561 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     7621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     8221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     8281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     8341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     8401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat
     8461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     9241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     9301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     9361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
