LOCUS       sequence-147631-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human codon optimized plasmid to express arC9 t2a mCherry w/2xNLS 
            and a sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10073)
  AUTHORS   Oakes BL, Nadler DC, Flamholz A, Fellmann C, Staahl BT, Doudna JA, 
            Savage DF
  TITLE     Profiling of engineering hotspots identifies an allosteric 
            CRISPR-Cas9 switch.
  JOURNAL   Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528.
  PUBMED    27136077
REFERENCE   2  (bases 1 to 10073)
  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: "Nat 
            Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528."
COMMENT     Sequence Label: pBLO1808_arC9_2xNLS_human
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        651..899
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     651..671
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     822..841
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        1100..1385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1387..1664
                     /label=chicken beta-actin promoter
     intron          1665..1893
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1905..1914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1914..1979
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1986..2006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4407..6905
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
                     SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
                     AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH
                     DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
                     NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
                     QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
                     VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
                     LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
                     GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
                     ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL
                     PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
                     RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP
                     SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
                     KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL
                     IHQSITGLYETRIDLSQLGGD"
     CDS             6906..6953
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6975..7028
                     /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             7029..7739
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7174..7192)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(7764..7781)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7770..7977
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7986..8126
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7986..8115
                     /label=AAV2 ITR
     rep_origin      8201..8656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8288..8307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8498..8519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8673..8692)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8792..8814
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8852..8870)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8938..9042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9043..9111,9112..9903)
                     /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(9261..9280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
      661 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
      721 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
      781 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
      841 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
      901 ggtcttcgag aagacctgtt taagagctat gctggaaaca gcatagcaag tttaaataag
      961 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
     1021 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
     1081 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     1141 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
     1201 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     1261 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
     1321 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     1381 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
     1441 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
     1501 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
     1561 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     1621 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
     1681 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     1741 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1801 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
     1861 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata
     1921 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga
     1981 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt
     2041 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     2101 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     2161 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     2221 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     2281 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     2341 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     2401 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     2461 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     2521 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     2581 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     2641 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     2701 tgatcgccca gctgcccggc gcatctccag caaacagtct tgccttgagt ctcacagcag
     2761 accagatggt gtcagcactg ctcgatgcgg agccacctat actctactct gaatacgacc
     2821 caaccagacc gttcagcgag gcttcaatga tgggactcct gactaatctg gcggacaggg
     2881 aacttgtgca catgatcaat tgggccaaac gggttcccgg ttttgttgac cttaccctgc
     2941 atgatcaggt tcatctgttg gaatgtgcat ggctcgaaat acttatgatt gggctcgtat
     3001 ggaggtccat ggagcaccct ggcaagctcc tgtttgctcc aaatctcttg ttggatcgga
     3061 atcagggtaa atgtgtcgaa ggaatggtag agatttttga tatgctcctg gcaacctcca
     3121 gtcggttccg aatgatgaat ctgcaggggg aggagttcgt gtgtctgaaa tctattatcc
     3181 tgcttaattc cggcgtttat acttttctgt catctacact gaagtccctg gaggagaagg
     3241 accacattca tcgcgtgttg gacaagataa ccgacacact catccatctg atggctaagg
     3301 cagggctgac cctgcaacag caacaccaga ggcttgccca actgttgctc attctgagcc
     3361 acatcaggca tatgtccaat aaaggaatgg aacacctgta cagcatgaag tgtaagaacg
     3421 tagtgccact ctatgatctt cttttggaga tgctggacgc tcacaggctc cacgcaccac
     3481 ctgcatctcc aggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg
     3541 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     3601 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     3661 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     3721 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     3781 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     3841 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     3901 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     3961 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     4021 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     4081 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     4141 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     4201 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     4261 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     4321 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     4381 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     4441 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     4501 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     4561 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     4621 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     4681 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     4741 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     4801 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     4861 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     4921 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     4981 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     5041 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     5101 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     5161 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     5221 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     5281 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     5341 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     5401 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     5461 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     5521 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     5581 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     5641 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     5701 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     5761 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     5821 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     5881 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     5941 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     6001 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     6061 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     6121 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     6181 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     6241 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     6301 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     6361 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     6421 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     6481 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     6541 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     6601 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     6661 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     6721 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     6781 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     6841 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     6901 gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aagggatccg
     6961 aatccggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc
     7021 ctggcccaat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     7081 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     7141 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     7201 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     7261 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     7321 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     7381 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     7441 gccccgtaat gcagaagaaa accatgggct gggaggcctc ctccgagcgg atgtaccccg
     7501 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     7561 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     7621 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     7681 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
     7741 aattcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     7801 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     7861 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     7921 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
     7981 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     8041 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     8101 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     8161 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     8221 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     8281 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     8341 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     8401 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     8461 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     8521 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     8581 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     8641 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     8701 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     8761 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     8821 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     8881 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     8941 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     9001 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     9061 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     9121 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     9181 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     9241 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     9301 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     9361 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     9421 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     9481 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     9541 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     9601 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     9661 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     9721 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
     9781 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     9841 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     9901 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     9961 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    10021 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
