LOCUS       sequence-391656-        9367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct encoding the high specificity SpCas9-KARA-D10A 
            nickase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9367)
  AUTHORS   Wang Q, Liu J, Janssen JM, Le Bouteiller M, Frock RL, Goncalves MAFV
  TITLE     Precise and broad scope genome editing based on high-specificity 
            Cas9 nickases.
  JOURNAL   Nucleic Acids Res. 2021 Jan 25;49(2):1173-1198. doi: 
            10.1093/nar/gkaa1236.
  PUBMED    33398349
REFERENCE   2  (bases 1 to 9367)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2021-01-25- 25"; volume: "49"; issue: 
            "2"; pages: "1173-1198"
COMMENT     Sequence Label: AP70_pU.CAG.Cas9-D10A-K848A-R1060A.rBGpA
FEATURES             Location/Qualifiers
     source          1..9367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        33..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..689
                     /label=chicken beta-actin promoter
     intron          690..1706
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1630..1652
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1714..1733
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1845..1854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1854..1919
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1926..1946
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1971..4508,4509..4511,4512..4973,4974..4976,4977..5144,
                     5145..5147,5148..6071)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6072..6119
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6234..6253)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6299..6354
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6353..6372)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6733..6750)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6733..6749)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6742..6764)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6958..6977)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7077..7099
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7137..7155)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7223..7327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7328..7396,7397..8188)
                     /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(7546..7565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8359..8947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8848..8867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9101..9118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9235..9256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9271..9276,9277..9294,9295..9301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9309..9325
                     /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)."
     primer_bind     9314..9336
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9333..9349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9333..9349
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtttaaacat ttaaatctcg agccatggat tcgacattga ttattgacta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
      421 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
      481 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggsgcgcgc
      541 caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag
      601 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
      661 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
      721 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
      781 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac
      841 ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt
      901 gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg
      961 gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc
     1021 gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg
     1081 aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg
     1141 cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1201 cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1261 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1321 aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt
     1381 gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg
     1441 agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg
     1501 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc
     1561 ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg
     1621 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg
     1681 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     1741 atcattttgg caaagaatta tcgcatgcct gcagagctct agagtctaat gtttaattac
     1801 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata
     1861 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga
     1921 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt
     1981 acagcatcgg cctggctatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     2041 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     2101 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     2161 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     2221 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     2281 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     2341 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     2401 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     2461 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     2521 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     2581 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     2641 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     2701 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     2761 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     2821 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     2881 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     2941 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     3001 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     3061 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     3121 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     3181 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     3241 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     3301 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     3361 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     3421 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     3481 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     3541 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     3601 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     3661 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     3721 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     3781 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     3841 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     3901 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     3961 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     4021 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     4081 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     4141 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     4201 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     4261 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     4321 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     4381 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     4441 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
     4501 gctttctggc cgacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     4561 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     4621 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     4681 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     4741 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     4801 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     4861 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     4921 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     4981 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     5041 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     5101 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaaggcgcct ctgatcgaga
     5161 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     5221 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     5281 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     5341 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     5401 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
     5461 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     5521 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     5581 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
     5641 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
     5701 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
     5761 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
     5821 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
     5881 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
     5941 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
     6001 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
     6061 tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagt
     6121 aagaattggc cgcacttaag ttacgcgtgg aaattcactc ctcaggtgca ggctgcctat
     6181 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     6241 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     6301 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     6361 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     6421 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     6481 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     6541 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     6601 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     6661 cctcttctct tatggagatc cctcgacctg cagcccaagc tgatcccggg atttaaatgt
     6721 ttaaacgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     6781 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     6841 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     6901 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     6961 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     7021 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     7081 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     7141 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     7201 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     7261 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     7321 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     7381 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     7441 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     7501 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7561 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7621 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7681 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7741 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7801 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7861 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7921 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7981 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     8041 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     8101 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     8161 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     8221 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8281 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8341 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     8401 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8461 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     8521 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8581 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8641 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8701 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8761 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8821 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8881 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8941 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     9001 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     9061 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     9121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     9181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     9241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     9301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     9361 caagctt
//
