LOCUS       sequence-169778-       14810 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral vector for the delivery of multiple sgRNAs targeting 
            different genes with constitutive Cas9 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 14810)
  AUTHORS   Cao J, Wu L, Zhang SM, Lu M, Cheung WK, Cai W, Gale M, Xu Q, Yan Q
  TITLE     An easy and efficient inducible CRISPR/Cas9 platform with improved 
            specificity for multiple gene targeting.
  JOURNAL   Nucleic Acids Res. 2016 Nov 2;44(19):e149. Epub 2016 Jul 25.
  PUBMED    27458201
REFERENCE   2  (bases 1 to 14810)
  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: "Nucleic 
            Acids Res."; date: "2016-11-2"; volume: "44(19)"; pages: "e149. Epub
            2016 Jul 25"
COMMENT     Sequence Label: Lenti-multi-CRISPR
FEATURES             Location/Qualifiers
     source          1..14810
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        110..157
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             176..550
                     /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    680..801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(717..736)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     771..790
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(850..866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(850..866)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(863..885)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    874..890
                     /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(898..904,905..922,923..928))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    943..964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1081..1098)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1252..1840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1332..1351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2011..2802,2803..2871))
                     /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     2634..2653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2872..2976)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3051..3070)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3242..3621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3623..3821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3776..3796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3839..4019
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4066..4191
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4684..4917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5102..5146
                     /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    5444..5561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5612..5860
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        join(5884..5889,5890..5907,5908..5914)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5968..6627
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6034..6053)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             6969..7274
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     7153..7172
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        7313..7388
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7681..7929
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7938..8013
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8075..8286
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8305..8314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8311..12414
                     /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             8311..12414
                     /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             12415..12462
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12463..12486
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12496..12552
                     /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             12553..13149
                     /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     13046..13066
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    13165..13753
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13218..13238)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13636..13647)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             13825..14058
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(14084..14101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    14090..14314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      14360..14788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14447..14466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14657..14678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
       61 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
      121 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
      181 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
      241 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
      301 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
      361 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
      421 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
      481 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
      541 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
      601 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
      661 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
      721 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
      781 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
      841 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      901 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      961 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1021 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1081 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1141 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1201 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1261 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1321 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1381 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1441 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1501 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1561 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1621 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1681 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1741 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1801 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1861 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1921 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1981 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2041 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2101 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2161 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2221 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2281 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2341 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2401 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2461 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2521 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2581 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2641 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2701 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2761 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2821 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2881 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2941 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3001 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     3061 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     3121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     3181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     3241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     3301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     3361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     3421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     3481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     3541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     3601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     3661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     3721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     3781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
     3841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     3901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     3961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     4021 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     4081 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     4141 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     4201 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     4261 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     4321 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     4381 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     4441 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     4501 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     4561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     4621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     4681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     4741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     4801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     4861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     4921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     4981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     5041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     5101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     5161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     5221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     5281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     5341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     5401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     5461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     5521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     5581 gcagagatcc agtttggtta attaaggtac cgagggccta tttcccatga ttccttcata
     5641 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
     5701 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     5761 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     5821 tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagacgatt aggcacccca
     5881 ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg
     5941 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     6001 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     6061 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     6121 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     6181 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     6241 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     6301 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     6361 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     6421 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     6481 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     6541 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     6601 tgcgatgagt ggcagggcgg ggcgtaaaga tctggatccg gcttactaaa agccagataa
     6661 cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc
     6721 cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg
     6781 acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac
     6841 catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg
     6901 gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg
     6961 ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     7021 tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca
     7081 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     7141 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
     7201 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     7261 tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcagct gggttttaga
     7321 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     7381 gtcggtgctt ttttaagctt ggcgtaacta gatcttgaga caaatggcag tattcatcca
     7441 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagag tccggcaagt
     7501 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     7561 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     7621 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccaga
     7681 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     7741 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     7801 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     7861 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     7921 cgaaacaccg cgtctctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     7981 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg aattcgctag ctaggtcttg
     8041 aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     8101 cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag gtggcgcggg
     8161 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     8221 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     8281 acacaggacc ggttctagag cgctgccacc atggacaaga agtacagcat cggcctggac
     8341 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     8401 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     8461 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     8521 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     8581 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     8641 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     8701 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     8761 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     8821 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     8881 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     8941 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     9001 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     9061 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     9121 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     9181 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     9241 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     9301 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     9361 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     9421 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     9481 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     9541 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     9601 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     9661 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     9721 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     9781 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     9841 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     9901 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     9961 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
    10021 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
    10081 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
    10141 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
    10201 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
    10261 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
    10321 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
    10381 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
    10441 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
    10501 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
    10561 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
    10621 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
    10681 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
    10741 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
    10801 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
    10861 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
    10921 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
    10981 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
    11041 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
    11101 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
    11161 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
    11221 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
    11281 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
    11341 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
    11401 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
    11461 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
    11521 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
    11581 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
    11641 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
    11701 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
    11761 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
    11821 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
    11881 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
    11941 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
    12001 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
    12061 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
    12121 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
    12181 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
    12241 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
    12301 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
    12361 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaagcga
    12421 cctgccgcca caaagaaggc tggacaggct aagaagaaga aagattacaa agacgatgac
    12481 gataagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag
    12541 aatcctggac cgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
    12601 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
    12661 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
    12721 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
    12781 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
    12841 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
    12901 cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag
    12961 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
    13021 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
    13081 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
    13141 ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
    13201 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
    13261 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
    13321 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
    13381 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
    13441 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
    13501 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
    13561 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
    13621 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
    13681 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
    13741 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
    13801 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
    13861 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
    13921 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
    13981 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
    14041 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
    14101 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
    14161 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
    14221 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
    14281 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
    14341 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
    14401 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
    14461 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
    14521 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
    14581 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
    14641 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
    14701 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
    14761 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
//
