LOCUS       sequence-372951-       12607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PspCas13b in mammalian cells for knockdown of target RNAs 
            in combination with compatible guide RNAs. Localized to the 
            cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12607)
  AUTHORS   Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung 
            J, Zhang F
  TITLE     RNA editing with CRISPR-Cas13.
  JOURNAL   Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
  PUBMED    29070703
REFERENCE   2  (bases 1 to 12607)
  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: "Science. 
            2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180."
COMMENT     Sequence Label: pC0046-EF1a-PspCas13b-NES-HIV
FEATURES             Location/Qualifiers
     source          1..12607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1574..1600
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1610..1690
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     misc_feature    1718..2306
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1771..1791)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2189..2200)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2309..2325)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2310..2326)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2831..3011
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3037..3054)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3043..3267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3313..3741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3400..3419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3610..3631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3750..3770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3755..4084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3935..4070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3997..4016
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4132..4179
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4198..4572
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4702..4823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4739..4758)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4793..4812
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4872..4888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4872..4888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4885..4907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4896..4912
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4920..4926,4927..4944,4945..4950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4965..4986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5103..5120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5274..5862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5354..5373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6033..6824,6825..6893))
                     /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     6656..6675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6894..6998)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7073..7092)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7264..7643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7645..7843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7798..7818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7861..8041
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8088..8213
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8706..8939
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9124..9168
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    9466..9583
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        9708..10886
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          9939..10877
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     10834..10854
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gagaatcaag agattcaagg acgaccggaa gtccattcgg agcgccgaca acaagatggg
       61 aaagagaggc ttcaagcaga tctccacagg caagctggcc gacttcctgg ccaaggacat
      121 cgtgctgttt cagcccagcg tgaacgatgg cgagaacaag atcaccggcc tgaactaccg
      181 gatcatgcag agcgccattg ccgtgtacga tagcggcgac gattacgagg ccaagcagca
      241 gttcaagctg atgttcgaga aggcccggct gatcggcaag ggcacaacag agcctcatcc
      301 atttctgtac aaggtgttcg cccgcagcat ccccgccaat gccgtcgagt tctacgagcg
      361 ctacctgatc gagcggaagt tctacctgac cggcctgtcc aacgagatca agaaaggcaa
      421 cagagtggat gtgcccttca tccggcggga ccagaacaag tggaaaacac ccgccatgaa
      481 gaccctgggc agaatctaca gcgaggatct gcccgtggaa ctgcccagac agatgttcga
      541 caatgagatc aagtcccacc tgaagtccct gccacagatg gaaggcatcg acttcaacaa
      601 tgccaacgtg acctatctga tcgccgagta catgaagaga gtgctggacg acgacttcca
      661 gaccttctac cagtggaacc gcaactaccg gtacatggac atgcttaagg gcgagtacga
      721 cagaaagggc tccctgcagc actgcttcac cagcgtggaa gagagagaag gcctctggaa
      781 agagcgggcc tccagaacag agcggtacag aaagcaggcc agcaacaaga tccgcagcaa
      841 ccggcagatg agaaacgcca gcagcgaaga gatcgagaca atcctggata agcggctgag
      901 caacagccgg aacgagtacc agaaaagcga gaaagtgatc cggcgctaca gagtgcagga
      961 tgccctgctg tttctgctgg ccaaaaagac cctgaccgaa ctggccgatt tcgacggcga
     1021 gaggttcaaa ctgaaagaaa tcatgcccga cgccgagaag ggaatcctga gcgagatcat
     1081 gcccatgagc ttcaccttcg agaaaggcgg caagaagtac accatcacca gcgagggcat
     1141 gaagctgaag aactacggcg acttctttgt gctggctagc gacaagagga tcggcaacct
     1201 gctggaactc gtgggcagcg acatcgtgtc caaagaggat atcatggaag agttcaacaa
     1261 atacgaccag tgcaggcccg agatcagctc catcgtgttc aacctggaaa agtgggcctt
     1321 cgacacatac cccgagctgt ctgccagagt ggaccgggaa gagaaggtgg acttcaagag
     1381 catcctgaaa atcctgctga acaacaagaa catcaacaaa gagcagagcg acatcctgcg
     1441 gaagatccgg aacgccttcg atcacaacaa ttaccccgac aaaggcgtgg tggaaatcaa
     1501 ggccctgcct gagatcgcca tgagcatcaa gaaggccttt ggggagtacg ccatcatgaa
     1561 gggatccctt caactgcctc cacttgaaag actgacactg ggtagttcct acccatacga
     1621 tgttccagat tacgcttatc cctacgacgt gcctgattat gcatacccat atgatgtccc
     1681 cgactatgcc taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     1741 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1801 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     1861 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     1921 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     1981 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2041 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2101 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     2161 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2221 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     2281 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca
     2341 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     2401 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     2461 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     2521 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     2581 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     2641 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     2701 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     2761 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     2821 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     2881 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     2941 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     3001 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     3061 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3121 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3181 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3241 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3301 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3421 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3601 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3721 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3781 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3901 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3961 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4021 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4081 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     4141 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     4201 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     4261 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     4321 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     4381 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     4441 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     4501 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     4561 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4621 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4681 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4741 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4801 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4861 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4921 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4981 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5041 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5101 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5161 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5221 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5281 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5341 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5401 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5461 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5521 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5581 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5641 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5701 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5761 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5821 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5881 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5941 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6001 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6061 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6121 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6181 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6241 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6301 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6361 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6421 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6481 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6541 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6601 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6661 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6721 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6781 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6841 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6901 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6961 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7021 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     7081 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7141 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7201 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     7261 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     7321 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     7381 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     7441 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     7501 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     7561 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     7621 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     7681 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     7741 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     7801 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
     7861 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7921 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7981 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     8041 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     8101 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     8161 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     8221 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     8281 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     8341 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     8401 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     8461 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     8521 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     8581 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     8641 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     8701 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     8761 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     8821 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     8881 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     8941 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     9001 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     9061 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     9121 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     9181 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     9241 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     9301 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     9361 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     9421 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     9481 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     9541 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     9601 cagcagagat ccagtttggt taattaatgc aaagatggat aaagttttaa acagagagga
     9661 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
     9721 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     9781 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     9841 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     9901 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     9961 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
    10021 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
    10081 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
    10141 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
    10201 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
    10261 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
    10321 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
    10381 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
    10441 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
    10501 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
    10561 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
    10621 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
    10681 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
    10741 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
    10801 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
    10861 tttttcttcc atttcaggtg tcgtgacgta cggccaccat gaacatcccc gctctggtgg
    10921 aaaaccagaa gaagtacttt ggcacctaca gcgtgatggc catgctgaac gctcagaccg
    10981 tgctggacca catccagaag gtggccgata ttgagggcga gcagaacgag aacaacgaga
    11041 atctgtggtt tcaccccgtg atgagccacc tgtacaacgc caagaacggc tacgacaagc
    11101 agcccgagaa aaccatgttc atcatcgagc ggctgcagag ctacttccca ttcctgaaga
    11161 tcatggccga gaaccagaga gagtacagca acggcaagta caagcagaac cgcgtggaag
    11221 tgaacagcaa cgacatcttc gaggtgctga agcgcgcctt cggcgtgctg aagatgtaca
    11281 gggacctgac caaccactac aagacctacg aggaaaagct gaacgacggc tgcgagttcc
    11341 tgaccagcac agagcaacct ctgagcggca tgatcaacaa ctactacaca gtggccctgc
    11401 ggaacatgaa cgagagatac ggctacaaga cagaggacct ggccttcatc caggacaagc
    11461 ggttcaagtt cgtgaaggac gcctacggca agaaaaagtc ccaagtgaat accggattct
    11521 tcctgagcct gcaggactac aacggcgaca cacagaagaa gctgcacctg agcggagtgg
    11581 gaatcgccct gctgatctgc ctgttcctgg acaagcagta catcaacatc tttctgagca
    11641 ggctgcccat cttctccagc tacaatgccc agagcgagga acggcggatc atcatcagat
    11701 ccttcggcat caacagcatc aagctgccca aggaccggat ccacagcgag aagtccaaca
    11761 agagcgtggc catggatatg ctcaacgaag tgaagcggtg ccccgacgag ctgttcacaa
    11821 cactgtctgc cgagaagcag tcccggttca gaatcatcag cgacgaccac aatgaagtgc
    11881 tgatgaagcg gagcagcgac agattcgtgc ctctgctgct gcagtatatc gattacggca
    11941 agctgttcga ccacatcagg ttccacgtga acatgggcaa gctgagatac ctgctgaagg
    12001 ccgacaagac ctgcatcgac ggccagacca gagtcagagt gatcgagcag cccctgaacg
    12061 gcttcggcag actggaagag gccgagacaa tgcggaagca agagaacggc accttcggca
    12121 acagcggcat ccggatcaga gacttcgaga acatgaagcg ggacgacgcc aatcctgcca
    12181 actatcccta catcgtggac acctacacac actacatcct ggaaaacaac aaggtcgaga
    12241 tgtttatcaa cgacaaagag gacagcgccc cactgctgcc cgtgatcgag gatgatagat
    12301 acgtggtcaa gacaatcccc agctgccgga tgagcaccct ggaaattcca gccatggcct
    12361 tccacatgtt tctgttcggc agcaagaaaa ccgagaagct gatcgtggac gtgcacaacc
    12421 ggtacaagag actgttccag gccatgcaga aagaagaagt gaccgccgag aatatcgcca
    12481 gcttcggaat cgccgagagc gacctgcctc agaagatcct ggatctgatc agcggcaatg
    12541 cccacggcaa ggatgtggac gccttcatca gactgaccgt ggacgacatg ctgaccgaca
    12601 ccgagcg
//
