LOCUS       sequence-423143-        9111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for transient Cas9 and EGFP expression. Small tRNA promoter 
            for sgRNA cloning by GoldenGate..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9111)
  TITLE     Chromosome 8p-engineering reveals increased metastasis targetable by
            patient-specific synthetic lethality
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9111)
  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            "Unpublished"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: pX458-tRNA-SpCas9
FEATURES             Location/Qualifiers
     source          1..9111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        106..181
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        278..563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        565..842
                     /label=chicken beta-actin promoter
     intron          843..1070
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1082..1091
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1091..1156
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1163..1183
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1208..3745,3746..3748,3749..4210,4211..4213,4214..4381,
                     4382..4384,4385..5308)
                     /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             5309..5356
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5372..5425
                     /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(5426..5428,5429..6139)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5426..5428,5429..6139)
                     /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(5729..5748)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6076..6097
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6167..6184)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6173..6380
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6389..6529
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6389..6518
                     /label=AAV2 ITR
     rep_origin      6604..7059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6691..6710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6901..6922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7076..7095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7195..7217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7255..7273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7341..7445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7446..7514,7515..8306)
                     /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(7664..7683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8477..9065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8966..8985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgtggttc catggtgtaa tggttagcac tctggactct gaatccagcg
       61 atccgagttc aaatctcggt ggaacctggg tcttcgagaa gacctgtttt agagctagaa
      121 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      181 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
      241 tgcgccaatt ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
      361 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      421 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      481 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      541 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      601 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      661 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      721 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      781 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      841 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      901 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      961 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
     1021 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
     1081 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
     1141 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
     1201 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     1261 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     1321 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     1381 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     1441 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     1501 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     1561 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     1621 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     1681 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     1741 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     1801 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     1861 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     1921 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     1981 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     2041 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     2101 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     2161 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     2221 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     2281 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     2341 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     2401 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     2461 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     2521 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     2581 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     2641 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     2701 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     2761 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     2821 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     2881 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     2941 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     3001 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     3061 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     3121 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     3181 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     3241 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     3301 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     3361 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     3421 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     3481 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     3541 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     3601 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     3661 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     3721 ccatatcgtg cctcagagct ttctggcgga cgactccatc gacaacaagg tgctgaccag
     3781 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     3841 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     3901 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     3961 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     4021 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     4081 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     4141 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     4201 aaagtaccct gcgctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     4261 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     4321 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     4381 ggcgcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     4441 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     4501 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     4561 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     4621 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     4681 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     4741 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     4801 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     4861 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     4921 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     4981 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     5041 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     5101 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     5161 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     5221 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     5281 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     5341 ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga ggaagtctgc taacatgcgg
     5401 tgacgtcgag gagaatcctg gcccagtgag caagggcgag gagctgttca ccggggtggt
     5461 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     5521 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     5581 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     5641 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5701 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5761 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5821 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     5881 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     5941 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     6001 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     6061 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     6121 catggacgag ctgtacaagg aattctaact agagctcgct gatcagcctc gactgtgcct
     6181 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6241 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6301 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag
     6361 aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc
     6421 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     6481 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     6541 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     6601 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6661 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6721 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6781 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     6841 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6901 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     6961 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     7021 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     7081 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     7141 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7201 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     7261 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     7321 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7381 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7441 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7501 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7561 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7621 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7681 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7741 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7801 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7861 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7921 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7981 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     8041 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     8101 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8161 gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8221 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8281 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8341 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     8401 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8461 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8521 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8581 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     9001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     9061 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg c
//
