LOCUS       sequence-381120-        9204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressing sgRNA-2 targeting eIF3f locus closed to stop coden.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9204)
  AUTHORS   Duan H, Zhang S, Zarai Y, Ollinger R, Wu Y, Sun L, Hu C, He Y, Tian 
            G, Rad R, Kong X, Cheng Y, Tuller T, Wolf DA
  TITLE     eIF3 mRNA selectivity profiling reveals eIF3k as a cancer-relevant 
            regulator of ribosome content.
  JOURNAL   EMBO J. 2023 Jun 15;42(12):e112362. doi: 10.15252/embj.2022112362. 
            Epub 2023 May 8.
  PUBMED    37155573
REFERENCE   2  (bases 1 to 9204)
  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: "EMBO J."; 
            date: "2023-06-15"; volume: "42(12):e112362. doi"; pages: " 
            10.15252/embj.2022112362. Epub 2023 May 8"
COMMENT     Sequence Label: eIF3f sgRNA-2  PX459 plasmid
FEATURES             Location/Qualifiers
     source          1..9204
                     /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        298..373
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        470..755
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1035..1262
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1274..1283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1283..1348
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1355..1375
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1400..4798,4799..4801,4802..5398,5399..5401,5402..5404,
                     5405..5407,5408..5500)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5501..5548
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5564..5617
                     /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             5618..6217
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5618..5637)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6114..6134
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6245..6262)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6251..6458
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6467..6607
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6467..6596
                     /label=AAV2 ITR
     rep_origin      6682..7137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6769..6788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6979..7000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7154..7173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7273..7295
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7333..7351)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7419..7523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7524..7592,7593..8384)
                     /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(7742..7761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8555..9143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9044..9063
                     /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 gccaacctca cacagtcacg tttcaccggc caacctcaca cagtcacgtt
      301 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
      361 accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc
      421 gtttttagcg cgtgcgccaa ttctgcagac aaatggctct agaggtaccc gttacataac
      481 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag
      541 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      601 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      661 gtaaatggcc cgcctggcat tgtgcccagt acatgacctt atgggacttt cctacttggc
      721 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      781 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      841 tattttgtgc agcgatgggg gcgggggggg ggggggggsg sgcgcsgggg gggggggggg
      901 gggggggggg gggggggggg gggggggggg ggggkgsggc ggcagccaat cagagcggcg
      961 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
     1021 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
     1081 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
     1141 acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt
     1201 tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc
     1261 aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga
     1321 tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
     1381 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
     1441 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     1501 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     1561 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     1621 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     1681 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     1741 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     1801 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     1861 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     1921 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     1981 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     2041 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     2101 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     2161 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     2221 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     2281 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     2341 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     2401 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     2461 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     2521 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     2581 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     2641 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     2701 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     2761 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     2821 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     2881 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     2941 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     3001 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     3061 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     3121 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     3181 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     3241 tctggaagat atagtgttga ctcttaccct ctttgaagat cgggaaatga ttgaggaaag
     3301 actaaaaaca tacgctcacc tgttcgacga taaggttatg aaacagttaa agaggcgtcg
     3361 ctatacgggc tggggacgat tgtcgcggaa acttatcaac gggataagag acaagcaaag
     3421 tggtaaaact attctcgatt ttctaaagag cgacggcttc gccaatagga actttatgca
     3481 gctgatccat gatgactctt taaccttcaa agaggatata caaaaggcac aggtttccgg
     3541 acaaggggac tcattgcacg aacatattgc gaatcttgct ggttcgccag ccatcaaaaa
     3601 gggcatactc cagacagtca aagtagtgga tgagctagtt aaggtcatgg gacgtcacaa
     3661 accggaaaac attgtaatcg agatggcacg cgaaaatcaa acgactcaga aggggcaaaa
     3721 aaacagtcga gagcggatga agagaataga agagggtatt aaagaactgg gcagccagat
     3781 cttaaaggag catcctgtgg aaaataccca attgcagaac gagaaacttt acctctatta
     3841 cctacaaaat ggaagggaca tgtatgttga tcaggaactg gacataaacc gtttatctga
     3901 ttacgacgtc gatcacattg taccccaatc ctttttgaag gacgattcaa tcgacaataa
     3961 agtgcttaca cgctcggata agaaccgagg gaaaagtgac aatgttccaa gcgaggaagt
     4021 cgtaaagaaa atgaagaact attggcggca gctcctaaat gcgaaactga taacgcaaag
     4081 aaagttcgat aacttaacta aagctgagag gggtggcttg tctgaacttg acaaggccgg
     4141 atttattaaa cgtcagctcg tggaaacccg ccaaatcaca aagcatgttg cacagatact
     4201 agattcccga atgaatacga aatacgacga gaacgataag ctgattcggg aagtcaaagt
     4261 aatcacttta aagtcaaaat tggtgtcgga cttcagaaag gattttcaat tctataaagt
     4321 tagggagata aataactacc accatgcgca cgacgcttat cttaatgccg tcgtagggac
     4381 cgcactcatt aagaaatacc cgaagctaga aagtgagttt gtgtatggtg attacaaagt
     4441 ttatgacgtc cgtaagatga tcgcgaaaag cgaacaggag ataggcaagg ctacagccaa
     4501 atacttcttt tattctaaca ttatgaattt ctttaagacg gaaatcactc tggcaaacgg
     4561 agagatacgc aaacgacctt taattgaaac caatggggag acaggtgaaa tcgtatggga
     4621 taagggccgg gacttcgcga cggtgagaaa agttttgtcc atgccccaag tcaacatagt
     4681 aaagaaaact gaggtgcaga ccggagggtt ttcaaaggaa tcgattcttc caaaaaggaa
     4741 tagtgataag ctcatcgctc gtaaaaagga ctgggacccg aaaaagtacg gtggcttcgt
     4801 gagccctaca gttgcctatt ctgtcctagt agtggcaaaa gttgagaagg gaaaatccaa
     4861 gaaactgaag tcagtcaaag aattattggg gataacgatt atggagcgct cgtcttttga
     4921 aaagaacccc atcgacttcc ttgaggcgaa aggttacaag gaagtaaaaa aggatctcat
     4981 aattaaacta ccaaagtata gtctgtttga gttagaaaat ggccgaaaac ggatgttggc
     5041 tagcgccgga gagcttcaaa aggggaacga actcgcacta ccgtctaaat acgtgaattt
     5101 cctgtattta gcgtcccatt acgagaagtt gaaaggttca cctgaagata acgaacagaa
     5161 gcaacttttt gttgagcagc acaaacatta tctcgacgaa atcatagagc aaatttcgga
     5221 attcagtaag agagtcatcc tagctgatgc caatctggac aaagtattaa gcgcatacaa
     5281 caagcacagg gataaaccca tacgtgagca ggcggaaaat attatccatt tgtttactct
     5341 taccaacctc ggcgctccag ccgcattcaa gtattttgac acaacgatag atcgcaaaca
     5401 gtacagatct accaaggagg tgctagacgc gacactgatt caccaatcca tcacgggatt
     5461 atatgaaact cggatagatt tgtcacagct tgggggtgac aaaaggccgg cggccacgaa
     5521 aaaggccggc caggcaaaaa agaaaaagga attcggcagt ggagagggca gaggaagtct
     5581 gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg accgagtaca agcccacggt
     5641 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     5701 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     5761 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     5821 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     5881 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5941 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     6001 cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     6061 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     6121 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     6181 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagaa ttctaactag agctcgctga
     6241 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     6301 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6361 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     6421 ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg
     6481 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     6541 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     6601 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6661 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6721 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     6781 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6841 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     6901 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6961 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     7021 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     7081 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     7141 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     7201 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     7261 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     7321 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     7381 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7441 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7501 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     7561 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7621 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7681 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     7741 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7801 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7861 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7921 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7981 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     8041 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     8101 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     8161 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     8221 ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc
     8281 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     8341 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     8401 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     8461 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8521 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8581 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8641 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8701 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8761 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8821 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8881 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8941 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     9001 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     9061 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     9121 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     9181 tttgctggcc ttttgctcac atgt
//
