LOCUS       sequence-313305-       15536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct with Cas9, Cre recombinase and U6 driven sgRNA 
            cassette with modified tracr RNA (20bp sgRNA can be cloned by 
            digesting the stuffer 1.8kb with BsmBI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15536)
  AUTHORS   Loganathan SK, Schleicher K, Malik A, Quevedo R, Langille E, Teng K,
            Oh RH, Rathod B, Tsai R, Samavarchi-Tehrani P, Pugh TJ, Gingras AC, 
            Schramek D
  TITLE     Rare driver mutations in head and neck squamous cell carcinomas 
            converge on NOTCH signaling.
  JOURNAL   Science. 2020 Mar 13;367(6483):1264-1269. doi: 
            10.1126/science.aax0902.
  PUBMED    32165588
REFERENCE   2  (bases 1 to 15536)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2020-03-13- 13"; volume: "367"; 
            issue: "6483"; pages: "1264-1269"
COMMENT     Sequence Label: pCRISPR-Cre stuffer v4
FEATURES             Location/Qualifiers
     source          1..15536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        4979..5190
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5198..5207
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5207..5272
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5279..5299
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5324..9424
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5324..9424
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9425..9472
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9488..9541
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9548..9568
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    10628..11216
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10681..10701)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11099..11110)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11288..11521
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11547..11564)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11553..11777
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11823..12251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11910..11929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12120..12141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12260..12280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12265..12594
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12445..12580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12507..12526
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12642..12689
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12708..13082
                     /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    13212..13333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13249..13268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13303..13322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13382..13398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13382..13398)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13395..13417)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13406..13422
                     /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(13430..13436,13437..13454,13455..13460))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13475..13496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13613..13630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13784..14372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13864..13883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14543..15334,15335..15403))
                     /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     15166..15185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15404..15508)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggag acggttgtaa atgagcacac
     2881 aaaatacaca tgctaaaata ttatattcta tgacctttat aaaatcaacc aaaatcttct
     2941 ttttaataac tttagtatca ataattagaa tttttatgtt cctttttgca aacttttaat
     3001 aaaaatgagc aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt cttcaccttt
     3061 aataatagct actccaccac ttgttcctaa gcggtcagct cctgcttcaa tcattttttg
     3121 agcatcttca aatgttctaa ctccaccagc tgctttaact aaagcattgt ctttaacaac
     3181 tgacttcatt agtttaacat cttcaaatgt tgcacctgat tttgaaaatc ctgttgatgt
     3241 tttaacaaat tctaatccag cttcaacagc tatttcacaa gctttcatga tttcttcttt
     3301 tgttaataaa caattttcca taatacattt aacaacatgt gatccagctg ctttttttac
     3361 agctttcatg tcttctaaaa ctaattcata atttttgtct tttaatgcac caatatttaa
     3421 taccatatca atttctgttg caccatcttt aattgcttca gaaacttcga atgcttttgt
     3481 agctgttgtg catgcaccta gaggaaaacc tacaacattt gttattccta catttgtgcc
     3541 ttttaataat tctttacaat agcttgttca atatgaatta acacaaactg ttgcaaaatc
     3601 aaattcaatt gcttcatcac ataattgttt aatttcagct ttcgtagcat cttgttttaa
     3661 taatgtgtga tctatatatt tgtttagttt cattttttct cctatatatt catttttaat
     3721 tttaattctt taataatttc gtctacttta actttagcgt tttgaacaga ttcaccaaca
     3781 cctataaaat aaatttttag tttaggttca gttccacttg ggcgaacagc aaatcatgac
     3841 ttatcttcta aataaaattt tagtaagtct tgtcctggca tattatacat tccatcgatg
     3901 tagtcttcaa cattaacaac tttaagtcca gcaatttgag ttaagggtgt tgctctcaat
     3961 gatttcatta atggttcaat ttttaatttc ttttcttctg gtttaaaatt caagtttaaa
     4021 gtgaaagtgt aatatgcacc catttcttta aataaatctt ctaaatagtc tactaatgtt
     4081 ttattttgtt ttttataaaa tcaagcagcc tctgctatta atatagaagc ttgtattcca
     4141 tctttatctc tagctgagtc atcaattaca tatccataac tttcttcata agcaaaaaca
     4201 aaatttaatc cgttatcttc ttctttagca atttctctac ccattcattt aaatccagtt
     4261 aaagcgcgca caatattaat tctatcaccc gcgtcacttg ttacaaaact tgaatataga
     4321 gccggatttt ttggagcatc tgggtcattc ataataataa tatctgcatc ttcaaatgca
     4381 tgctcttcaa cctcaataac gttatatcct gattcacgta atatttttgg gtaaatttag
     4441 ttcctgttcc attaactgcg ctggatccgt acgtctctgt ttaagagcta tgctggaaac
     4501 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     4561 gtgctttttt gaattcgcta gctaggtctt gaaaggagtg ggaaatccag ttaaagcgcg
     4621 cacaatatta attctatcac ccgcgtcact tgttacaaaa cttgaatata gagccggatt
     4681 ttttggagca tctgggtcat tcataataat aatatctgca tcttcaaatg catgctcttc
     4741 aacctcaata acgttatatc ctgattcacg taatattttt ggggtaaatt tagttcctgt
     4801 tccattaact gcgctggatc cgtacgtctc tgtttaagag ctatgctgga aacagcatag
     4861 caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     4921 tttgaattcg ctagctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     4981 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc
     5041 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     5101 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     5161 tttcgcaacg ggtttgccgc cagaacacag gaccggtgcc accatggact ataaggacca
     5221 cgacggagac tacaaggatc atgatattga ttacaaagac gatgacgata agatggcccc
     5281 aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat
     5341 cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     5401 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     5461 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     5521 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     5581 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     5641 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     5701 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     5761 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     5821 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     5881 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     5941 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     6001 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
     6061 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     6121 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     6181 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     6241 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     6301 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     6361 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     6421 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     6481 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     6541 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     6601 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     6661 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     6721 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     6781 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     6841 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     6901 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     6961 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     7021 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     7081 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     7141 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     7201 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     7261 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     7321 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     7381 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     7441 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     7501 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     7561 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     7621 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     7681 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     7741 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     7801 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
     7861 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
     7921 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     7981 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     8041 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     8101 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     8161 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     8221 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     8281 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     8341 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     8401 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     8461 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     8521 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     8581 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     8641 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     8701 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
     8761 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     8821 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     8881 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     8941 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
     9001 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     9061 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     9121 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     9181 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     9241 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     9301 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
     9361 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     9421 cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa aggaattcgg
     9481 cagtggagag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc
     9541 aatggtgccc aagaagaaga ggaaagtctc caacctgctg actgtgcacc aaaacctgcc
     9601 tgccctccct gtggatgcca cctctgatga agtcaggaag aacctgatgg acatgttcag
     9661 ggacaggcag gccttctctg aacacacctg gaagatgctc ctgtctgtgt gcagatcctg
     9721 ggctgcctgg tgcaagctga acaacaggaa atggttccct gctgaacctg aggatgtgag
     9781 ggactacctc ctgtacctgc aagccagagg cctggctgtg aagaccatcc aacagcacct
     9841 gggccagctc aacatgctgc acaggagatc tggcctgcct cgcccttctg actccaatgc
     9901 tgtgtccctg gtgatgagga gaatcagaaa ggagaatgtg gatgctgggg agagagccaa
     9961 gcaggccctg gcctttgaac gcactgactt tgaccaagtc agatccctga tggagaactc
    10021 tgacagatgc caggacatca ggaacctggc cttcctgggc attgcctaca acaccctgct
    10081 gcgcattgcc gaaattgcca gaatcagagt gaaggacatc tcccgcaccg atggtgggag
    10141 aatgctgatc cacattggca ggaccaagac cctggtgtcc acagctggtg tggagaaggc
    10201 cctgtccctg ggggttacca agctggtgga gagatggatc tctgtgtctg gtgtggctga
    10261 tgaccccaac aactacctgt tctgccgggt cagaaagaat ggtgtggctg ccccttctgc
    10321 cacctcccaa ctgtccaccc gggccctgga agggatcttt gaggccaccc accgcctgat
    10381 ctatggtgcc aaggatgact ctgggcagag atacctggcc tggtctggcc actctgccag
    10441 agtgggtgct gccagggaca tggccagggc tggtgtgtcc atccctgaaa tcatgcaggc
    10501 tggtggctgg accaatgtga acattgtgat gaactacatc agaaacctgg actctgagac
    10561 tggggccatg gtgaggcttc tagaggatgg ggattgagtc gacgaattcg atatcaagct
    10621 tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
    10681 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
    10741 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
    10801 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
    10861 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
    10921 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
    10981 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
    11041 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
    11101 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
    11161 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
    11221 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
    11281 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
    11341 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
    11401 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
    11461 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
    11521 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
    11581 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
    11641 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
    11701 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
    11761 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
    11821 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
    11881 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
    11941 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
    12001 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
    12061 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
    12121 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
    12181 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
    12241 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
    12301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
    12361 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
    12421 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
    12481 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
    12541 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
    12601 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
    12661 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
    12721 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
    12781 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
    12841 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
    12901 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
    12961 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
    13021 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
    13081 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
    13141 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
    13201 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    13261 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    13321 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
    13381 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
    13441 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
    13501 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
    13561 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
    13621 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
    13681 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
    13741 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
    13801 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
    13861 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
    13921 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
    13981 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
    14041 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
    14101 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
    14161 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
    14221 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
    14281 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
    14341 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
    14401 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
    14461 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
    14521 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
    14581 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
    14641 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
    14701 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
    14761 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
    14821 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
    14881 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
    14941 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
    15001 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    15061 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    15121 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    15181 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    15241 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    15301 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    15361 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    15421 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    15481 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
