LOCUS       sequence-230492-        8900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for human codon optimized 
            AsCas12a-enRVR(E174R/S542R/K548V/N552R) nuclease with C-terminal NLS
            and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8900)
  AUTHORS   Kleinstiver BP, Sousa AA, Walton RT, Tak YE, Hsu JY, Clement K, 
            Welch MM, Horng JE, Malagon-Lopez J, Scarfo I, Maus MV, Pinello L, 
            Aryee MJ, Joung JK
  TITLE     Engineered CRISPR-Cas12a variants with increased activities and 
            improved targeting ranges for gene, epigenetic and base editing.
  JOURNAL   Nat Biotechnol. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 
            10.1038/s41587-018-0011-0.
  PUBMED    30742127
REFERENCE   2  (bases 1 to 8900)
  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. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 
            10.1038/s41587-018-0011-0."
COMMENT     Sequence Label: 
            pCAG-hAsCas12a-enRVR(E174R/S542R/K548V/N552R)-NLS(nucleoplasmin)-3xH
            A (AAS1060)
FEATURES             Location/Qualifiers
     source          1..8900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          160..1176
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1100..1122
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1184..1203
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1275..5195
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLARGWDVNVEKNRGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      4915..4924
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5196..5243
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5250..5330
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(5420..5439)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5485..5540
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5539..5558)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5901..5917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5901..5917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5914..5936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5925..5941
                     /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(5949..5955,5956..5973,5974..5979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5994..6015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6074..6269
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6120..6255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6275..6409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6312..6331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6366..6385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6477..6494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6648..7236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6728..6747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7407..8198,8199..8267))
                     /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     8030..8049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8268..8372)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        8403..8782
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggsgcgcgc crggsggggs ggggsggggc gaggggsggg gsggggsgrg gsggagaggt
       61 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      121 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      181 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      241 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      301 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      361 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
      421 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
      481 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
      541 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
      601 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
      661 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
      721 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
      781 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
      841 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
      901 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
      961 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1021 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1081 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1141 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1201 gtgctgtctc atcattttgg caaagaattc tgcagtcgac ggtaccgcgg gcccgggatc
     1261 caccggtcgc caccatgaca cagttcgagg gctttaccaa cctgtatcag gtgagcaaga
     1321 cactgcggtt tgagctgatc ccacagggca agaccctgaa gcacatccag gagcagggct
     1381 tcatcgagga ggacaaggcc cgcaatgatc actacaagga gctgaagccc atcatcgatc
     1441 ggatctacaa gacctatgcc gaccagtgcc tgcagctggt gcagctggat tgggagaacc
     1501 tgagcgccgc catcgactcc tatagaaagg agaaaaccga ggagacaagg aacgccctga
     1561 tcgaggagca ggccacatat cgcaatgcca tccacgacta cttcatcggc cggacagaca
     1621 acctgaccga tgccatcaat aagagacacg ccgagatcta caagggcctg ttcaaggccg
     1681 agctgtttaa tggcaaggtg ctgaagcagc tgggcaccgt gaccacaacc gagcacgaga
     1741 acgccctgct gcggagcttc gacaagttta caacctactt ctccggcttt tatagaaaca
     1801 ggaagaacgt gttcagcgcc gaggatatca gcacagccat cccacaccgc atcgtgcagg
     1861 acaacttccc caagtttaag gagaattgtc acatcttcac acgcctgatc accgccgtgc
     1921 ccagcctgcg ggagcacttt gagaacgtga agaaggccat cggcatcttc gtgagcacct
     1981 ccatcgagga ggtgttttcc ttcccttttt ataaccagct gctgacacag acccagatcg
     2041 acctgtataa ccagctgctg ggaggaatct ctcgggaggc aggcaccgag aagatcaagg
     2101 gcctgaacga ggtgctgaat ctggccatcc agaagaatga tgagacagcc cacatcatcg
     2161 cctccctgcc acacagattc atccccctgt ttaagcagat cctgtccgat aggaacaccc
     2221 tgtctttcat cctggaggag tttaagagcg acgaggaagt gatccagtcc ttctgcaagt
     2281 acaagacact gctgagaaac gagaacgtgc tggagacagc cgaggccctg tttaacgagc
     2341 tgaacagcat cgacctgaca cacatcttca tcagccacaa gaagctggag acaatcagca
     2401 gcgccctgtg cgaccactgg gatacactga ggaatgccct gtatgagcgg agaatctccg
     2461 agctgacagg caagatcacc aagtctgcca aggagaaggt gcagcgcagc ctgaagcacg
     2521 aggatatcaa cctgcaggag atcatctctg ccgcaggcaa ggagctgagc gaggccttca
     2581 agcagaaaac cagcgagatc ctgtcccacg cacacgccgc cctggatcag ccactgccta
     2641 caaccctgaa gaagcaggag gagaaggaga tcctgaagtc tcagctggac agcctgctgg
     2701 gcctgtacca cctgctggac tggtttgccg tggatgagtc caacgaggtg gaccccgagt
     2761 tctctgcccg gctgaccggc atcaagctgg agatggagcc ttctctgagc ttctacaaca
     2821 aggccagaaa ttatgccacc aagaagccct actccgtgga gaagttcaag ctgaactttc
     2881 agatgcctac actggccaga ggctgggacg tgaatgtgga gaagaacaga ggcgccatcc
     2941 tgtttgtgaa gaacggcctg tactatctgg gcatcatgcc aaagcagaag ggcaggtata
     3001 aggccctgag cttcgagccc acagagaaaa ccagcgaggg ctttgataag atgtactatg
     3061 actacttccc tgatgccgcc aagatgatcc caaagtgcag cacccagctg aaggccgtga
     3121 cagcccactt tcagacccac acaaccccca tcctgctgtc caacaatttc atcgagcctc
     3181 tggagatcac aaaggagatc tacgacctga acaatcctga gaaggagcca aagaagtttc
     3241 agacagccta cgccaagaaa accggcgacc agaagggcta cagagaggcc ctgtgcaagt
     3301 ggatcgactt cacaagggat tttctgtcca agtataccaa gacaacctct atcgatctgt
     3361 ctagcctgcg gccatcctct cagtataagg acctgggcga gtactatgcc gagctgaatc
     3421 ccctgctgta ccacatcagc ttccagagaa tcgccgagaa ggagatcatg gatgccgtgg
     3481 agacaggcaa gctgtacctg ttccagatct ataacaagga ctttgccaag ggccaccacg
     3541 gcaagcctaa tctgcacaca ctgtattgga ccggcctgtt ttctccagag aacctggcca
     3601 agacaagcat caagctgaat ggccaggccg agctgttcta ccgccctaag tccaggatga
     3661 agaggatggc acaccggctg ggagagaaga tgctgaacaa gaagctgaag gatcagaaaa
     3721 ccccaatccc cgacaccctg taccaggagc tgtacgacta tgtgaatcac agactgtccc
     3781 acgacctgtc tgatgaggcc agggccctgc tgcccaacgt gatcaccaag gaggtgtctc
     3841 acgagatcat caaggatagg cgctttacca gcgacaagtt ctttttccac gtgcctatca
     3901 cactgaacta tcaggccgcc aattccccat ctaagttcaa ccagagggtg aatgcctacc
     3961 tgaaggagca ccccgagaca cctatcatcg gcatcgatcg gggcgagaga aacctgatct
     4021 atatcacagt gatcgactcc accggcaaga tcctggagca gcggagcctg aacaccatcc
     4081 agcagtttga ttaccagaag aagctggaca acagggagaa ggagagggtg gcagcaaggc
     4141 aggcctggtc tgtggtgggc acaatcaagg atctgaagca gggctatctg agccaggtca
     4201 tccacgagat cgtggacctg atgatccact accaggccgt ggtggtgctg gagaacctga
     4261 atttcggctt taagagcaag aggaccggca tcgccgagaa ggccgtgtac cagcagttcg
     4321 agaagatgct gatcgataag ctgaattgcc tggtgctgaa ggactatcca gcagagaaag
     4381 tgggaggcgt gctgaaccca taccagctga cagaccagtt cacctccttt gccaagatgg
     4441 gcacccagtc tggcttcctg ttttacgtgc ctgccccata tacatctaag atcgatcccc
     4501 tgaccggctt cgtggacccc ttcgtgtgga aaaccatcaa gaatcacgag agccgcaagc
     4561 acttcctgga gggcttcgac tttctgcact acgacgtgaa aaccggcgac ttcatcctgc
     4621 actttaagat gaacagaaat ctgtccttcc agaggggcct gcccggcttt atgcctgcat
     4681 gggatatcgt gttcgagaag aacgagacac agtttgacgc caagggcacc cctttcatcg
     4741 ccggcaagag aatcgtgcca gtgatcgaga atcacagatt caccggcaga taccgggacc
     4801 tgtatcctgc caacgagctg atcgccctgc tggaggagaa gggcatcgtg ttcagggatg
     4861 gctccaacat cctgccaaag ctgctggaga atgacgattc tcacgccatc gacaccatgg
     4921 tggccctgat ccgcagcgtg ctgcagatgc ggaactccaa tgccgccaca ggcgaggact
     4981 atatcaacag ccccgtgcgc gatctgaatg gcgtgtgctt cgactcccgg tttcagaacc
     5041 cagagtggcc catggacgcc gatgccaatg gcgcctacca catcgccctg aagggccagc
     5101 tgctgctgaa tcacctgaag gagagcaagg atctgaagct gcagaacggc atctccaatc
     5161 aggactggct ggcctacatc caggagctgc gcaacaaaag gccggcggcc acgaaaaagg
     5221 ccggccaggc aaaaaagaaa aagggatcct acccatacga tgttccagat tacgcttatc
     5281 cctacgacgt gcctgattat gcatacccat atgatgtccc cgactatgcc taaagcggcc
     5341 gcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     5401 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     5461 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     5521 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     5581 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     5641 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     5701 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     5761 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     5821 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     5881 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6061 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6121 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6181 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6241 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     6301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     6421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg
     8401 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     8461 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     8521 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     8581 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     8641 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     8701 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     8761 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
     8821 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
     8881 gatgggggcg gggggggggg
//
