LOCUS       sequence-334794-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 and guide RNA for the site neighbouring KLF7 gene..
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-KLF7-1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..234
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     189..209
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             252..432
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    479..604
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1097..1330
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1515..1559
                     /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    1857..1974
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2025..2273
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2025..2045
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2196..2215
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2295..2370
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2432..2643
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2662..2671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2668..6771
                     /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             2668..6771
                     /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             6772..6819
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6820..6843
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6853..6909
                     /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             6910..7506
                     /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     7403..7423
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7522..8110
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7575..7595)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7993..8004)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8182..8415
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8441..8458)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8447..8671
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8717..9145
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8804..8823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9014..9035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9154..9174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9159..9488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9339..9474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9401..9420
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9536..9583
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9602..9976
                     /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    10106..10227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10143..10162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10197..10216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10276..10292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10276..10292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10289..10311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10300..10316
                     /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(10324..10330,10331..10348,10349..10354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10369..10390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10507..10524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10678..11266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10758..10777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11437..12228,12229..12297))
                     /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     12060..12079
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12298..12402)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12477..12496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(12668..13013,1..34)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
       61 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      121 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      181 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
      241 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
      301 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
      361 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
      421 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
      481 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
      541 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
      601 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
      661 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
      721 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
      781 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
      841 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
      901 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
      961 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1021 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1081 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1141 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1201 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1261 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1321 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1381 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1441 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1501 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     1561 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1621 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     1681 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     1741 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     1801 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     1861 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     1921 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     1981 tattacaggg acagcagaga tccagtttgg ttaattaagg taccgagggc ctatttccca
     2041 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt
     2101 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     2161 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     2221 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgatcact
     2281 ccagctcgtg gatcgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     2341 caacttgaaa aagtggcacc gagtcggtgc ttttttgaat tcgctagcta ggtcttgaaa
     2401 ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     2461 gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta
     2521 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     2581 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     2641 caggaccggt tctagagcgc tgccaccatg gacaagaagt acagcatcgg cctggacatc
     2701 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     2761 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     2821 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     2881 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     2941 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     3001 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     3061 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     3121 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     3181 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     3241 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     3301 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     3361 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac
     3421 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     3481 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     3541 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     3601 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     3661 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     3721 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     3781 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     3841 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     3901 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     3961 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     4021 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     4081 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     4141 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     4201 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     4261 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     4321 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     4381 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     4441 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     4501 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     4561 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     4621 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     4681 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     4741 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     4801 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     4861 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     4921 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     4981 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     5041 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     5101 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     5161 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc
     5221 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     5281 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     5341 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     5401 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
     5461 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
     5521 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
     5581 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
     5641 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
     5701 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
     5761 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
     5821 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
     5881 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
     5941 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
     6001 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
     6061 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
     6121 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
     6181 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
     6241 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
     6301 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
     6361 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
     6421 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
     6481 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
     6541 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
     6601 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
     6661 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
     6721 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga caagcgacct
     6781 gccgccacaa agaaggctgg acaggctaag aagaagaaag attacaaaga cgatgacgat
     6841 aagggatccg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat
     6901 cctggaccga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     6961 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     7021 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     7081 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     7141 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     7201 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     7261 aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt
     7321 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     7381 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     7441 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     7501 gcctgaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     7561 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     7621 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     7681 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     7741 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     7801 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     7861 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     7921 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     7981 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     8041 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     8101 gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact
     8161 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     8221 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     8281 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     8341 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     8401 ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     8461 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     8521 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     8581 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     8641 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     8701 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     8761 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     8821 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     8881 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     8941 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     9001 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     9061 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     9121 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     9181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     9241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     9301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     9361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     9421 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     9481 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     9541 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     9601 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     9661 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     9721 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     9781 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     9841 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     9901 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     9961 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
    10021 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
    10081 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
    10141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    10201 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
    10261 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    10321 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    10381 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    10441 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    10501 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    10561 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    10621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    10681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    10741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    10801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    10861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    10921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    10981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    11041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    11101 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    11161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    11221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    11281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    11341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    11401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    11461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    11521 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    11581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    11641 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    11701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    11761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    11821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    11881 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    11941 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    12001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    12061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    12121 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    12181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    12241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    12301 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    12361 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    12421 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
    12481 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
    12541 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
    12601 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
    12661 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
    12721 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
    12781 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
    12841 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
    12901 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
    12961 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gca
//
