LOCUS       sequence-450810-        8511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CBh-SaCas9-2A-Puro, and 2X hU6-sgRNA (Sa) with BbsI golden gate 
            cloning backbone for dual gRNAs. sgRNA uses 3TC scaffold for 
            increased sgRNA expression. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8511)
  AUTHORS   Chey YCJ, Gierus L, Lushington C, Arudkumar JC, B Geiger A, Staker 
            LG, Robertson LJ, Pfitzner C, Kennedy JG, Lee RHB, Godahewa GI, 
            Adikusuma F, Thomas PQ
  TITLE     Optimal SpCas9- and SaCas9-mediated gene editing by enhancing gRNA 
            transcript levels through scaffold poly-T tract reduction.
  JOURNAL   BMC Genomics. 2025 Feb 12;26(1):138. doi: 
            10.1186/s12864-025-11317-2.
  PUBMED    39939860
REFERENCE   2  (bases 1 to 8511)
  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: "BMC 
            Genomics."; date: "2025-02-12"; volume: "26(1):138. doi"; pages: " 
            10.1186/s12864-025-11317-2"
COMMENT     Sequence Label: DG SaCas9 puro V3
FEATURES             Location/Qualifiers
     source          1..8511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          139..366
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      378..387
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             390..410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             435..3590
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             3591..3638
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3654..3707
                     /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             3708..4307
                     /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(3708..3727)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4204..4224
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4335..4352)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4341..4548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_RNA        complement(4654..4729)
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     promoter        complement(4747..4995)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     rep_origin      5078..5533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5165..5184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5375..5396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5550..5569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5669..5691
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5729..5747)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5815..5919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5920..5988,5989..6780)
                     /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(6138..6157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6951..7539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7440..7459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7601..7849
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7868..7943
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     enhancer        7964..8249
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
       61 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      121 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      181 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      241 cgggacggcc cttctcctcc gggctgtaat tagctgagca agaggtaagg gtttaaggga
      301 tggttggttg gtggggtatt aatgtttaat tacctggagc acctgcctga aatcactttt
      361 tttcaggttg gaccggtgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg
      421 gagtcccagc agccaagcgg aactacatcc tgggcctgga catcggcatc accagcgtgg
      481 gctacggcat catcgactac gagacacggg acgtgatcga tgccggcgtg cggctgttca
      541 aagaggccaa cgtggaaaac aacgagggca ggcggagcaa gagaggcgcc agaaggctga
      601 agcggcggag gcggcataga atccagagag tgaagaagct gctgttcgac tacaacctgc
      661 tgaccgacca cagcgagctg agcggcatca acccctacga ggccagagtg aagggcctga
      721 gccagaagct gagcgaggaa gagttctctg ccgccctgct gcacctggcc aagagaagag
      781 gcgtgcacaa cgtgaacgag gtggaagagg acaccggcaa cgagctgtcc accaaagagc
      841 agatcagccg gaacagcaag gccctggaag agaaatacgt ggccgaactg cagctggaac
      901 ggctgaagaa agacggcgaa gtgcggggca gcatcaacag attcaagacc agcgactacg
      961 tgaaagaagc caaacagctg ctgaaggtgc agaaggccta ccaccagctg gaccagagct
     1021 tcatcgacac ctacatcgac ctgctggaaa cccggcggac ctactatgag ggacctggcg
     1081 agggcagccc cttcggctgg aaggacatca aagaatggta cgagatgctg atgggccact
     1141 gcacctactt ccccgaggaa ctgcggagcg tgaagtacgc ctacaacgcc gacctgtaca
     1201 acgccctgaa cgacctgaac aatctcgtga tcaccaggga cgagaacgag aagctggaat
     1261 attacgagaa gttccagatc atcgagaacg tgttcaagca gaagaagaag cccaccctga
     1321 agcagatcgc caaagaaatc ctcgtgaacg aagaggatat taagggctac agagtgacca
     1381 gcaccggcaa gcccgagttc accaacctga aggtgtacca cgacatcaag gacattaccg
     1441 cccggaaaga gattattgag aacgccgagc tgctggatca gattgccaag atcctgacca
     1501 tctaccagag cagcgaggac atccaggaag aactgaccaa tctgaactcc gagctgaccc
     1561 aggaagagat cgagcagatc tctaatctga agggctatac cggcacccac aacctgagcc
     1621 tgaaggccat caacctgatc ctggacgagc tgtggcacac caacgacaac cagatcgcta
     1681 tcttcaaccg gctgaagctg gtgcccaaga aggtggacct gtcccagcag aaagagatcc
     1741 ccaccaccct ggtggacgac ttcatcctga gccccgtcgt gaagagaagc ttcatccaga
     1801 gcatcaaagt gatcaacgcc atcatcaaga agtacggcct gcccaacgac atcattatcg
     1861 agctggcccg cgagaagaac tccaaggacg cccagaaaat gatcaacgag atgcagaagc
     1921 ggaaccggca gaccaacgag cggatcgagg aaatcatccg gaccaccggc aaagagaacg
     1981 ccaagtacct gatcgagaag atcaagctgc acgacatgca ggaaggcaag tgcctgtaca
     2041 gcctggaagc catccctctg gaagatctgc tgaacaaccc cttcaactat gaggtggacc
     2101 acatcatccc cagaagcgtg tccttcgaca acagcttcaa caacaaggtg ctcgtgaagc
     2161 aggaagaaaa cagcaagaag ggcaaccgga ccccattcca gtacctgagc agcagcgaca
     2221 gcaagatcag ctacgaaacc ttcaagaagc acatcctgaa tctggccaag ggcaagggca
     2281 gaatcagcaa gaccaagaaa gagtatctgc tggaagaacg ggacatcaac aggttctccg
     2341 tgcagaaaga cttcatcaac cggaacctgg tggataccag atacgccacc agaggcctga
     2401 tgaacctgct gcggagctac ttcagagtga acaacctgga cgtgaaagtg aagtccatca
     2461 atggcggctt caccagcttt ctgcggcgga agtggaagtt taagaaagag cggaacaagg
     2521 ggtacaagca ccacgccgag gacgccctga tcattgccaa cgccgatttc atcttcaaag
     2581 agtggaagaa actggacaag gccaaaaaag tgatggaaaa ccagatgttc gaggaaaagc
     2641 aggccgagag catgcccgag atcgaaaccg agcaggagta caaagagatc ttcatcaccc
     2701 cccaccagat caagcacatt aaggacttca aggactacaa gtacagccac cgggtggaca
     2761 agaagcctaa tagagagctg attaacgaca ccctgtactc cacccggaag gacgacaagg
     2821 gcaacaccct gatcgtgaac aatctgaacg gcctgtacga caaggacaat gacaagctga
     2881 aaaagctgat caacaagagc cccgaaaagc tgctgatgta ccaccacgac ccccagacct
     2941 accagaaact gaagctgatt atggaacagt acggcgacga gaagaatccc ctgtacaagt
     3001 actacgagga aaccgggaac tacctgacca agtactccaa aaaggacaac ggccccgtga
     3061 tcaagaagat taagtattac ggcaacaaac tgaacgccca tctggacatc accgacgact
     3121 accccaacag cagaaacaag gtcgtgaagc tgtccctgaa gccctacaga ttcgacgtgt
     3181 acctggacaa tggcgtgtac aagttcgtga ccgtgaagaa tctggatgtg atcaaaaaag
     3241 aaaactacta cgaagtgaat agcaagtgct atgaggaagc taagaagctg aagaagatca
     3301 gcaaccaggc cgagtttatc gcctccttct acaacaacga tctgatcaag atcaacggcg
     3361 agctgtatag agtgatcggc gtgaacaacg acctgctgaa ccggatcgaa gtgaacatga
     3421 tcgacatcac ctaccgcgag tacctggaaa acatgaacga caagaggccc cccaggatca
     3481 ttaagacaat cgcctccaag acccagagca ttaagaagta cagcacagac attctgggca
     3541 acctgtatga agtgaaatct aagaagcacc ctcagatcat caaaaagggc aaaaggccgg
     3601 cggccacgaa aaaggccggc caggcaaaaa agaaaaagga attcggcagt ggagagggca
     3661 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg accgagtaca
     3721 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3781 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3841 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3901 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3961 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4021 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4081 tggccaccgt cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4141 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4201 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4261 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagaa ttctaactag
     4321 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4381 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4441 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4501 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga
     4561 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4621 gcgaccaaag gtcgcccgac gcccgggaaa aaatctcgcc aacaagttga cgagataaac
     4681 acggcatttt gccttgtttc agtagattct gtttccagag tactgaaacg tcttctcgaa
     4741 gaccccggtg tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca
     4801 agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc
     4861 aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa
     4921 attaattcca attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat
     4981 gggaaatagg ccctccctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     5041 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     5101 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     5161 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5221 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5281 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5341 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5401 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     5461 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5521 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5581 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5641 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5701 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5761 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5821 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5881 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5941 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6001 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6061 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6121 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6181 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6241 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6301 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6361 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6421 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6481 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6541 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6601 tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg
     6661 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6721 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6781 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6841 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6901 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6961 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7021 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7081 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7141 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7201 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7261 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7321 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7381 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7441 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7501 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7561 ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat ttcccatgat
     7621 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
     7681 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     7741 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     7801 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg ggtcttcgag
     7861 aagacctgtt tcagtactct ggaaacagaa tctactgaaa caaggcaaaa tgccgtgttt
     7921 atctcgtcaa cttgttggcg agattttttg cggccgcggt acccgttaca taacttacgg
     7981 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc
     8041 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     8101 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     8161 ggcccgcctg gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     8221 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     8281 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     8341 gtgcagcgat gggggcgggg gggggggggg ggcgcggcgc cnnggggggg gggggggggg
     8401 ngggnggggg ggggggggsg gaggkssksg rskkgnnngg ggrskgknsy gdnrrmtnng
     8461 scsssbgcns cgcwmngatw wragmnrsgg rkkkttaggt grksatrkrc a
//
