LOCUS       sequence-199157-       12862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PguCas13b 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 12862)
  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 12862)
  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: pC0045-EF1a-PguCas13b-NES-HIV
FEATURES             Location/Qualifiers
     source          1..12862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(145..162)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    151..375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      421..849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(508..527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     718..739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(858..878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        863..1192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1043..1178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1105..1124
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1240..1287
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1306..1680
                     /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    1810..1931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1847..1866)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1901..1920
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1980..1996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1980..1996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1993..2015)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2004..2020
                     /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(2028..2034,2035..2052,2053..2058))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2073..2094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2211..2228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2382..2970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2462..2481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3141..3932,3933..4001))
                     /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     3764..3783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4002..4106)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4181..4200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4372..4751
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4753..4951
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4906..4926
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4969..5149
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5196..5321
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5814..6047
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6232..6276
                     /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    6574..6691
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6816..7994
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7047..7985
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7942..7962
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             11544..11570
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             11580..11660
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     misc_feature    11688..12276
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11741..11761)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12159..12170)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12279..12295)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12280..12296)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             join(12801..12862,1..119)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
       61 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      121 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
      181 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
      241 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
      301 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
      361 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
      421 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      481 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      541 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      601 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      661 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      721 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      781 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      841 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
      901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
      961 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1021 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1081 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     1141 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     1201 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     1261 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     1321 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     1381 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     1441 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     1501 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     1561 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     1621 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     1681 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     1741 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     1801 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1861 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1921 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     1981 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2041 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2101 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2161 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2221 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2281 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2821 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3061 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3121 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3181 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3241 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3301 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3361 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3421 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3481 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3541 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3601 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3661 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3721 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3781 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3841 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3901 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3961 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4021 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4081 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     4141 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     4201 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     4261 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     4321 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     4381 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4441 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4501 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4561 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4621 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4681 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4741 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4801 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4861 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     4921 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
     4981 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5041 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5101 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     5161 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     5221 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     5281 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     5341 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     5401 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     5461 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     5521 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     5581 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     5641 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     5701 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     5761 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     5821 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     5881 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     5941 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     6001 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     6061 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     6121 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     6181 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     6241 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     6301 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     6361 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     6421 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     6481 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     6541 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     6601 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     6661 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     6721 agtttggtta attaatgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     6781 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
     6841 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     6901 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     6961 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     7021 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     7081 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
     7141 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
     7201 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
     7261 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
     7321 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
     7381 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
     7441 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
     7501 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
     7561 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
     7621 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
     7681 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     7741 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     7801 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     7861 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     7921 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     7981 ttcaggtgtc gtgacgtacg gccaccatga cagagcagag cgagaggccc tacaacggca
     8041 cctactacac cctggaagat aagcacttct gggccgcctt tctgaacctg gccagacaca
     8101 acgcctacat caccctgaca cacatcgacc ggcagctggc ctacagcaag gccgacatca
     8161 ccaacgacca ggacgtgctg agcttcaagg ccctgtggaa gaacttcgac aacgacctgg
     8221 aaagaaagag ccggctgcgg agcctgatcc tgaagcactt cagctttctg gaaggcgccg
     8281 cttacggcaa gaagctgttc gagtctaaga gcagcggcaa caagagcagc aagaacaaag
     8341 agctgaccaa gaaagagaaa gaggaactgc aggccaacgc tctgagcctg gacaacctga
     8401 agtctatcct gttcgacttc ctgcagaagc tgaaggactt ccggaactac tacagccact
     8461 accggcacag cggaagcagc gagctgcctc tgttcgacgg caacatgctg cagcggctgt
     8521 acaacgtgtt cgacgtgtcc gtgcagcgcg tgaagatcga ccacgagcac aacgatgagg
     8581 tggaccctca ctaccacttc aaccacctcg tgcggaaggg caagaaggac agatacggcc
     8641 acaacgacaa ccccagcttc aagcaccact tcgtggacgg cgaaggcatg gttacagaag
     8701 ccggcctgct gttcttcgtg tccctgttcc tggaaaagcg ggacgccatc tggatgcaga
     8761 agaagatccg gggcttcaaa ggcggcaccg agacatacca gcagatgacc aacgaggtgt
     8821 tctgccggtc cagaatcagc ctgcctaagc tgaagctgga aagcctgcgg atggacgact
     8881 ggatgctgct ggacatgctg aacgaactcg tgcggtgccc caagcctctg tacgacagac
     8941 tgagagagga cgaccgggcc tgctttagag tgcccgtgga tatcctgcct gacgaggacg
     9001 atacagatgg cggcggagag gaccccttca agaataccct cgtccggcac caggacagat
     9061 tcccttactt cgccctgcgg tacttcgacc tgaagaaggt gttcaccagc ctgcggttcc
     9121 acatcgatct gggcacctac cactttgcca tctacaagaa gatgatcggc gagcagcccg
     9181 aggacagaca cctgaccaga aacctgtacg gcttcggccg gatccaggac ttcgccgaag
     9241 aacacagacc cgaggaatgg aagcggctcg tcagagatct ggactacttc gagacaggcg
     9301 acaagcccta catcagccag acaagcccac actaccacat cgagaaggga aagatcggcc
     9361 tgagattcat gcccgagggc cagcatcttt ggccctctcc agaagtgggc accaccagaa
     9421 ccggcagatc taagtacgcc caggacaaga gactgaccgc cgaggccttt ctgtctgtgc
     9481 acgagctgat gcctatgatg ttctactact tcctgctgcg cgagaagtac agcgaggaag
     9541 tgtctgccga gagagtgcag ggcagaatca agcgcgtgat cgaggatgtg tacgccgtgt
     9601 acgatgcctt tgccagggac gagatcaaca cccgggatga gctggatgcc tgcctggccg
     9661 ataagggcat cagaagaggc catctgcctc gccagatgat cgccatcctg agccaagagc
     9721 acaaggacat ggaagagaag attcggaaga aactgcaaga gatgatggcc gacaccgacc
     9781 accggctgga tatgctggat agacagaccg accggaagat cagaatcggc cggaagaatg
     9841 ccggactgcc taagagcgga gtgatcgccg attggctcgt gcgggacatg atgagatttc
     9901 agcccgtggc caaggacgcc agcggaaagc ctctgaacaa ctccaaggcc aacagcaccg
     9961 agtaccggat gctgcagaga gccctggctc tgtttggagg cgagaaagag aggctgaccc
    10021 catacttccg gcagatgaat ctgaccggcg gaaacaaccc tcatcctttt ctgcacgaaa
    10081 ctcgctggga gagccacacc aacatcctgt ccttttaccg gtcctacctg agagcccgca
    10141 aggccttcct cgaaaggatc ggcagatccg acagagtgga aaaccggcca tttctgctgc
    10201 tgaaagagcc caagaccgac agacagacac tggtggctgg atggaagggc gagttccatc
    10261 tgccaagagg catcttcaca gaggccgtgc gggattgcct gatcgagatg ggacacgatg
    10321 aagtggccag ctacaaagaa gtgggattca tggccaaggc cgtgccactg tactttgaga
    10381 gagcctgcga ggacagggtg cagccctttt acgacagccc cttcaacgtg ggcaacagcc
    10441 tgaagcctaa gaagggcaga ttcctgagca aagaggaacg ggccgaagag tgggagcggg
    10501 gcaaagagag attccgggat ctcgaagcct ggtcctatag cgccgccaga agaatcgagg
    10561 acgcctttgc cggcatcgag tatgctagcc ccggaaacaa gaagaaaatc gagcagctcc
    10621 tgcgggacct gagcctgtgg gaagcctttg agagcaagct gaaagtgcgg gccgaccgga
    10681 tcaatctggc caagctgaag aaagaaatcc tggaagccca agaacacccc taccacgact
    10741 tcaagagctg gcagaagttc gagcgcgagc tgcggctggt caagaaccag gatatcatca
    10801 cctggatgat gtgccgcgac ctgatggaag aaaacaaggt ggaaggcctg gacaccggca
    10861 cactgtacct gaaggacatc agacccaacg tgcaagagca gggcagcctg aacgtgctga
    10921 acagagtgaa gcccatgaga ctgcccgtgg tggtgtacag agccgatagc agaggccacg
    10981 tgcacaaaga agaagcccct ctggccaccg tgtacatcga ggaaagagac accaagctgc
    11041 tgaagcaggg caacttcaag tccttcgtga aggacaggcg gctgaacggc ctgttctcct
    11101 tcgttgatac tggcggactg gccatggaac agtaccccat ctccaagctg agagtggaat
    11161 acgagctggc caagtaccag accgccagag tgtgtgtgtt cgagctgacc ctgagactgg
    11221 aagagtccct gctgacccgg tatcctcatc tgcccgacga gagcttcaga gagatgctgg
    11281 aatcttggag cgaccctctg ctggccaaat ggcctgaact gcacggaaaa gtgcggctgc
    11341 tgatcgccgt gcggaatgcc tttagccaca atcagtaccc tatgtacgac gaggccgtgt
    11401 tcagctccat ccggaagtac gatcctagca gccccgacgc catcgaagag agaatgggcc
    11461 tgaatatcgc ccaccgcctg tctgaggaag tgaagcaggc caaagaaacc gtggaacgga
    11521 tcattcaggc cggatccctt caactgcctc cacttgaaag actgacactg ggtagttcct
    11581 acccatacga tgttccagat tacgcttatc cctacgacgt gcctgattat gcatacccat
    11641 atgatgtccc cgactatgcc taagaattcg atatcaagct tatcgataat caacctctgg
    11701 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
    11761 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
    11821 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
    11881 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
    11941 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
    12001 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
    12061 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
    12121 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
    12181 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
    12241 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgagacc
    12301 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
    12361 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
    12421 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
    12481 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
    12541 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
    12601 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca
    12661 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
    12721 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag
    12781 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct
    12841 ctggctaact agggaaccca ct
//
