LOCUS       sequence-189971-       12002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for encoding a human codon-optimized SpCas9 driven
            by EFs promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12002)
  AUTHORS   Yao X, Wang X, Hu X, Liu Z, Liu J, Zhou H, Shen X, Wei Y, Huang Z, 
            Ying W, Wang Y, Nie YH, Zhang CC, Li S, Cheng L, Wang Q, Wu Y, Huang
            P, Sun Q, Shi L, Yang H
  TITLE     Homology-mediated end joining-based targeted integration using 
            CRISPR/Cas9.
  JOURNAL   Cell Res. 2017 May 19. doi: 10.1038/cr.2017.76.
  PUBMED    28524166
REFERENCE   2  (bases 1 to 12002)
  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: "Cell Res. 
            2017 May 19. doi: 10.1038/cr.2017.76."
COMMENT     Sequence Label: Lenti_Efs_hSpCas9_NLS_FLAG-WPRE
FEATURES             Location/Qualifiers
     source          1..12002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(277..705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(387..408)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     599..618
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    751..975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     964..981
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             1007..1240
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    1312..1900
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1418..1429
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1827..1847
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1916..1939)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1940..1987)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(1988..6091)
                     /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             complement(1988..6091)
                     /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"
     regulatory      6088..6097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6116..6327)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    6437..6554
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6852..6896)
                     /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    7081..7314
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7807..7932
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7979..8159
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8177..8375)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8202..8222)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        8377..8756
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     8928..8947
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        9022..9126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9127..9195,9196..9987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9345..9364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10158..10746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10647..10666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10900..10917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11034..11055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11070..11075,11076..11093,11094..11100)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11108..11124
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     11113..11135
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11132..11148
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11132..11148
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    11197..11318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11208..11227)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11262..11281
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(11448..11822)
                     /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"
     promoter        complement(11841..11888)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(11936..12002,1..263))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(11950..12002,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
      301 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      361 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
      421 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
      481 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
      541 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
      601 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
      661 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga taccccctag
      721 agccccagct ggttctttcc gcctcagaag ccatagagcc caccgcatcc ccagcatgcc
      781 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
      841 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
      901 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
      961 aactagaagg cacagtcgag gctgatcagc gggtttaaac gggccctgct agagattttc
     1021 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
     1081 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
     1141 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acaagcaaaa
     1201 agcagatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt ttcttttaaa
     1261 aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaagtcga cgcggggagg
     1321 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
     1381 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
     1441 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
     1501 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     1561 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
     1621 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
     1681 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     1741 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     1801 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     1861 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atcgatttag gatcccttat
     1921 cgtcatcgtc tttgtaatct ttcttcttct tagcctgtcc agccttcttt gtggcggcag
     1981 gtcgcttgtc gcctcccagc tgagacaggt cgatccgtgt ctcgtacagg ccggtgatgc
     2041 tctggtggat cagggtggcg tccagcacct ctttggtgct ggtgtacctc ttccggtcga
     2101 tggtggtgtc aaagtacttg aaggcggcag gggctcccag attggtcagg gtaaacaggt
     2161 ggatgatatt ctcggcctgc tctctgatgg gcttatcccg gtgcttgttg taggcggaca
     2221 gcactttgtc cagattagcg tcggccagga tcactctctt ggagaactcg ctgatctgct
     2281 cgatgatctc gtccaggtag tgcttgtgct gttccacaaa cagctgtttc tgctcattat
     2341 cctcggggga gcccttcagc ttctcatagt ggctggccag gtacaggaag ttcacatatt
     2401 tggagggcag ggccagttcg tttcccttct gcagttcgcc ggcagaggcc agcattctct
     2461 tccggccgtt ttccagctcg aacagggagt acttaggcag cttgatgatc aggtcctttt
     2521 tcacttcttt gtagcccttg gcttccagaa agtcgatggg attcttctcg aagctgcttc
     2581 tttccatgat ggtgatcccc agcagctctt tcacactctt cagtttcttg gacttgccct
     2641 tttccacttt ggccaccacc agcacagaat aggccacggt ggggctgtcg aagccgccgt
     2701 acttcttagg gtcccagtcc ttctttctgg cgatcagctt atcgctgttc ctcttgggca
     2761 ggatagactc tttgctgaag ccgcctgtct gcacctcggt ctttttcacg atattcactt
     2821 ggggcatgct cagcactttc cgcacggtgg caaaatcccg gcccttatcc cacacgatct
     2881 ccccggtttc gccgtttgtc tcgatcagag gccgcttccg gatctcgccg ttggccaggg
     2941 taatctcggt cttgaaaaag ttcatgatgt tgctgtagaa gaagtacttg gcggtagcct
     3001 tgccgatttc ctgctcgctc ttggcgatca tcttccgcac gtcgtacacc ttgtagtcgc
     3061 cgtacacgaa ctcgctttcc agcttagggt actttttgat cagggcggtt cccacgacgg
     3121 cgttcaggta ggcgtcgtgg gcgtggtggt agttgttgat ctcgcgcact ttgtaaaact
     3181 ggaaatcctt ccggaaatcg gacaccagct tggacttcag ggtgatcact ttcacttccc
     3241 ggatcagctt gtcattctcg tcgtacttag tgttcatccg ggagtccagg atctgtgcca
     3301 cgtgctttgt gatctgccgg gtttccacca gctgtctctt gatgaagccg gccttatcca
     3361 gttcgctcag gccgcctctc tcggccttgg tcagattgtc gaactttctc tgggtaatca
     3421 gcttggcgtt cagcagctgc cgccagtagt tcttcatctt cttcacgacc tcttcggagg
     3481 gcacgttgtc gctcttgccc cggttcttgt cgcttctggt cagcaccttg ttgtcgatgg
     3541 agtcgtcctt cagaaagctc tgaggcacga tatggtccac atcgtagtcg gacagccggt
     3601 tgatgtccag ttcctggtcc acgtacatat cccgcccatt ctgcaggtag tacaggtaca
     3661 gcttctcgtt ctgcagctgg gtgttttcca cggggtgttc tttcaggatc tggctgccca
     3721 gctctttgat gccctcttcg atccgcttca ttctctcgcg gctgttcttc tgtcccttct
     3781 gggtggtctg gttctctctg gccatttcga tcacgatgtt ctcgggcttg tgccggccca
     3841 tcactttcac gagctcgtcc accaccttca ctgtctgcag gatgcccttc ttaatggcgg
     3901 ggctgccggc cagattggca atgtgctcgt gcaggctatc gccctggccg gacacctggg
     3961 ctttctggat gtcctcttta aaggtcaggc tgtcgtcgtg gatcagctgc atgaagtttc
     4021 tgttggcgaa gccgtcggac ttcaggaaat ccaggattgt cttgccggac tgcttgtccc
     4081 ggatgccgtt gatcagcttc cggctcagcc tgccccagcc ggtgtatctc cgccgcttca
     4141 gctgcttcat cactttgtcg tcgaacaggt gggcataggt tttcagccgt tcctcgatca
     4201 tctctctgtc ctcaaacagt gtcagggtca gcacgatatc ttccagaatg tcctcgtttt
     4261 cctcattgtc caggaagtcc ttgtccttga taattttcag cagatcgtgg tatgtgccca
     4321 gggaggcgtt gaaccgatct tccacgccgg agatttccac ggagtcgaag cactcgattt
     4381 tcttgaagta gtcctctttc agctgcttca cggtcacttt ccggttggtc ttgaacagca
     4441 ggtccacgat ggcctttttc tgctcgccgc tcaggaaggc gggctttctc attccctcgg
     4501 tcacgtattt cactttggtc agctcgttat acacggtgaa gtactcgtac agcaggctgt
     4561 gcttgggcag caccttctcg ttgggcaggt tcttatcgaa gttggtcatc cgctcgatga
     4621 agctctgggc ggaagcgccc ttgtccacca cttcctcgaa gttccagggg gtgatggttt
     4681 cctcgctctt tctggtcatc caggcgaatc tgctgtttcc cctggccaga gggcccacgt
     4741 agtaggggat gcggaaggtc aggatcttct cgatcttttc ccggttgtcc ttcaggaatg
     4801 ggtaaaaatc ttcctgccgc cgcagaatgg cgtgcagctc tcccaggtgg atctggtggg
     4861 ggatgctgcc gttgtcgaag gtccgctgct tccgcagcag gtcctctctg ttcagcttca
     4921 cgagcagttc ctcggtgccg tccatctttt ccaggatggg cttgatgaac ttgtagaact
     4981 cttcctggct ggctccgccg tcaatgtagc cggcgtagcc gttcttgctc tggtcgaaga
     5041 aaatctcttt gtacttctca ggcagctgct gccgcacgag agctttcagc agggtcaggt
     5101 cctggtggtg ctcgtcgtat ctcttgatca tagaggcgct caggggggcc ttggtgatct
     5161 cggtgttcac tctcaggatg tcgctcagca ggatggcgtc ggacaggttc ttggcggcca
     5221 gaaacaggtc ggcgtactgg tcgccgatct gggccagcag gttgtccagg tcgtcgtcgt
     5281 aggtgtcctt gctcagctgc agtttggcat cctcggccag gtcgaagttg ctcttgaagt
     5341 tgggggtcag gcccaggctc agggcaatca ggtttccgaa caggccattc ttcttctcgc
     5401 cgggcagctg ggcgatcaga ttttccagcc gtctgctctt gctcagtctg gcagacagga
     5461 tggccttggc gtccacgccg ctggcgttga tggggttttc ctcgaacagc tggttgtagg
     5521 tctgcaccag ctggatgaac agcttgtcca cgtcgctgtt gtcggggttc aggtcgccct
     5581 cgatcaggaa gtggccccgg aacttgatca tgtgggccag ggccagatag atcagccgca
     5641 ggtcggcctt gtcggtgctg tccaccagtt tctttctcag gtggtagatg gtggggtact
     5701 tctcgtggta ggccacctcg tccacgatgt tgccgaagat ggggtgccgc tcgtgcttct
     5761 tatcctcttc caccaggaag gactcttcca gtctgtggaa gaagctgtcg tccaccttgg
     5821 ccatctcgtt gctgaagatc tcttgcagat agcagatccg gttcttccgt ctggtgtatc
     5881 ttcttctggc ggttctcttc agccgggtgg cctcggctgt ttcgccgctg tcgaacagca
     5941 gggctccgat caggttcttc ttgatgctgt gccggtcggt gttgcccagc accttgaatt
     6001 tcttgctggg caccttgtac tcgtcggtga tcacggccca gcccacagag ttggtgccga
     6061 tgtccaggcc gatgctgtac ttcttgtcca tggtggcagc gctctagaac cggtcctgtg
     6121 ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca cttatatacg
     6181 gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt tcccagttta
     6241 ccccgcgcca ccttctctag gcaccggatc aattgccgac ccctcccccc aacttctcgg
     6301 ggactgtggg cgatgtgcgc tctgcccact gacgggcacc ggagccaatt cccactcctt
     6361 tcaagaccta gctagcgaat tccgatggta ccttaattaa ccaaactgga tctctgctgt
     6421 ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt
     6481 gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc
     6541 cttttctttt aaaattgtgg atgaatactg ccatttgtct gcagaattgg cgcacgcagt
     6601 gccgatccgt tcactaatcg aatggatctg tctctgtctc tctctccacc ttcttcttct
     6661 attccttcgg gcctgtcggg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     6721 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     6781 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     6841 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     6901 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     6961 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     7021 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     7081 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     7141 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     7201 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     7261 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     7321 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     7381 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     7441 cctcctccag gtctgaagat cagcggccgc ttgctgtgcg gtggtcttac ttttgttttg
     7501 ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac
     7561 acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag
     7621 ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagg
     7681 attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt
     7741 tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt
     7801 aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg
     7861 cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct
     7921 gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg
     7981 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
     8041 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
     8101 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca
     8161 gtacaggcaa aacgcgctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     8221 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     8281 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     8341 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     8401 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     8461 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     8521 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     8581 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     8641 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     8701 atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaacg
     8761 cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt
     8821 catgcaattg tcggtcaagc cttgccttgt tgtagcttaa attttgctcg cgcactactc
     8881 agcgacctcc aacacacaag cagggagcag atactggctt aactatgcgg catcagagca
     8941 gattgtactg agagtgcacc ataggggatc gggagatctc ccgatccgtc gacgtcaggt
     9001 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     9061 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     9121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     9181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     9241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     9301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     9361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     9421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     9481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     9541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     9601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     9661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     9721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     9781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     9841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     9901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     9961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
    10021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
    10081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
    10141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
    10201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
    10261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
    10321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
    10381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
    10441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
    10501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
    10561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
    10621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
    10681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
    10741 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
    10801 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
    10861 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
    10921 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
    10981 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
    11041 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
    11101 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
    11161 aagctctagc tagaggtcga cggtatacag acatgataag atacattgat gagtttggac
    11221 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
    11281 ctttatttgt aaccattata agctgcaata aacaagttgg ggtgggcgaa gaactccagc
    11341 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
    11401 aacctttcat agaaggcggc ggtggaatcg aaatctcgta gcacgtgtca gtcctgctcc
    11461 tcggccacga agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac
    11521 ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg
    11581 acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg
    11641 ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc
    11701 acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac
    11761 tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc
    11821 atggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc cgatgattaa
    11881 ttgtcaacac gtgctgatca gatccgaaaa tggatataca agctcccggg agctttttgc
    11941 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg
    12001 cg
//
