LOCUS       sequence-334773-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 and guide RNA with untarget sequence generated by 
            scramble of the target sequence of LentiCRISPRv2-ACTB-C1...
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Yoshino Y, Endo S, Chen Z, Qi H, Watanabe G, Chiba N
  TITLE     Evaluation of site-specific homologous recombination activity of 
            BRCA1 by direct quantitation of gene editing efficiency.
  JOURNAL   Sci Rep. 2019 Feb 7;9(1):1644. doi: 10.1038/s41598-018-38311-x.
  PUBMED    30733539
REFERENCE   2  (bases 1 to 13013)
  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: "Sci Rep."; 
            date: "2019-02-7"; volume: "9(1):1644. doi"; pages: " 
            10.1038/s41598-018-38311-x"
COMMENT     Sequence Label: LentiCRISPRv2-scr
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..210
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     165..185
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1833..1950
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2001..2249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2001..2021
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2172..2191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2271..2346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2408..2619
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2638..2647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2644..6747
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2644..6747
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6748..6795
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6796..6819
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6829..6885
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6886..7482
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7379..7399
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7498..8086
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7551..7571)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7969..7980)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8158..8391
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8417..8434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8423..8647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8693..9121
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8780..8799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8990..9011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9130..9150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9135..9464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9315..9450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9377..9396
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9512..9559
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9578..9952
                     /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    10082..10203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10119..10138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10173..10192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10252..10268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10252..10268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10265..10287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10276..10292
                     /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(10300..10306,10307..10324,10325..10330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10345..10366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10483..10500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10654..11242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10734..10753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11413..12204,12205..12273))
                     /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     12036..12055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12274..12378)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12453..12472)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(12644..13013,1..10)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
       61 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      121 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      181 gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     1801 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     1861 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     1921 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     1981 gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac
     2041 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
     2101 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     2161 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
     2221 tatatatctt gtggaaagga cgaaacaccg cctgggttag agctaccgca gttttagagc
     2281 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     2341 cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc
     2401 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     2461 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     2521 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     2581 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc
     2641 accatggaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
     2701 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     2761 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     2821 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     2881 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     2941 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     3001 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     3061 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     3121 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     3181 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     3241 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     3301 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     3361 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     3421 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     3481 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     3541 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     3601 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     3661 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     3721 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     3781 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     3841 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     3901 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     3961 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     4021 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     4081 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     4141 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     4201 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     4261 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     4321 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     4381 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     4441 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     4501 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     4561 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     4621 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     4681 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     4741 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     4801 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     4861 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     4921 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     4981 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     5041 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     5101 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     5161 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     5221 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     5281 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     5341 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     5401 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     5461 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     5521 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     5581 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     5641 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     5701 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     5761 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     5821 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     5881 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     5941 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     6001 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     6061 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     6121 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     6181 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     6241 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     6301 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     6361 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     6421 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     6481 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     6541 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     6601 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     6661 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     6721 atcgacctgt ctcagctggg aggcgacaag cgacctgccg ccacaaagaa ggctggacag
     6781 gctaagaaga agaaagatta caaagacgat gacgataagg gatccggcgc aacaaacttc
     6841 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgaccga gtacaagccc
     6901 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     6961 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     7021 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     7081 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     7141 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     7201 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     7261 accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     7321 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     7381 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     7441 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat
     7501 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     7561 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     7621 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     7681 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     7741 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     7801 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     7861 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     7921 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     7981 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     8041 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     8101 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     8161 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     8221 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     8281 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     8341 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
     8401 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     8461 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     8521 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     8581 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     8641 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     8701 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     8761 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     8821 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     8881 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     8941 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     9001 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     9061 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     9121 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     9181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     9241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     9301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     9361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     9421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     9481 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     9541 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     9601 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     9661 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     9721 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     9781 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     9841 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     9901 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     9961 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
    10021 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
    10081 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    10141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
    10201 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
    10261 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    10321 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    10381 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    10441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    10501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    10561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    10621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    10681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    10741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    10801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    10861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    10921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    10981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    11041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    11101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    11161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    11221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    11281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    11341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    11401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    11461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    11521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    11581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    11641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    11701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    11761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    11821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    11881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    11941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    12001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    12061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    12121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    12181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    12241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    12301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    12361 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
    12421 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    12481 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
    12541 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
    12601 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
    12661 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
    12721 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
    12781 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
    12841 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
    12901 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
    12961 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgc
//
