LOCUS       sequence-418433-       13038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing SpCas9 and tracrRNA for stable cell 
            line extablishment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13038)
  AUTHORS   Zhang H, Kelly K, Lee J, Echeverria D, Cooper D, Panwala R, Amrani 
            N, Chen Z, Gaston N, Wagh A, Newby GA, Xie J, Liu DR, Gao G, Wolfe 
            SA, Khvorova A, Watts JK, Sontheimer EJ
  TITLE     Self-delivering, chemically modified CRISPR RNAs for AAV co-delivery
            and genome editing in vivo.
  JOURNAL   Nucleic Acids Res. 2023 Nov 30:gkad1125. doi: 10.1093/nar/gkad1125.
  PUBMED    38033325
REFERENCE   2  (bases 1 to 13038)
  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. 2023 Nov 30:gkad1125. doi: 10.1093/nar/gkad1125."
COMMENT     Sequence Label: pEJS1553 pLenti-U1a-SpCas9-miniU6-tracrRNA
FEATURES             Location/Qualifiers
     source          1..13038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             119..299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(325..342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    331..555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      601..1029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(688..707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     898..919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1038..1058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1203..1337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1264..1283
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1399..1446
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1465..1839
                     /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    1969..2090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2006..2025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2060..2079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2139..2155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2139..2155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2152..2174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2163..2179
                     /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(2187..2193,2194..2211,2212..2217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2232..2253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2370..2387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2541..3129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2621..2640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3300..4091,4092..4160))
                     /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     3923..3942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4161..4265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4340..4359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4531..4910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4912..5110
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5065..5085
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5128..5308
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5355..5480
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5973..6206
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6391..6435
                     /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    6733..6850
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     complement(7062..7082)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     regulatory      7418..7427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7424..11527
                     /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             7424..11527
                     /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             7424..11527
                     /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             11534..11560
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             11564..11620
                     /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             11621..12019
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    12044..12632
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12097..12117)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12515..12526)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12635..12651)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12636..12652)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
       61 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
      121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
      181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      301 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
      361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
      421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
      481 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
      541 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
      601 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      661 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      721 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      781 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      841 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      901 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      961 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1021 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     1141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     1201 agtcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     1261 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     1321 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     1381 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     1441 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     1501 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     1561 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     1621 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     1681 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     1741 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     1801 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     1861 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1921 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     1981 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2041 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     2101 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2161 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2221 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2281 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2341 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2941 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3241 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3421 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3601 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3901 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4261 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4321 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4381 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4441 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4501 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4561 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4621 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4681 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4741 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4801 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4861 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4921 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4981 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5041 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5101 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     5161 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5221 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5281 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     5341 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     5401 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     5461 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     5521 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     5581 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     5641 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     5701 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     5761 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     5821 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     5881 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     5941 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     6001 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     6061 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     6121 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     6181 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     6241 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     6301 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     6361 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     6421 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     6481 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     6541 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     6601 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     6661 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
     6721 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     6781 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     6841 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttagctagca
     6901 aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac
     6961 ttgctatgct cggtgtttcg tcctttccac aagatatata aagccaagaa atcgaaatac
     7021 tttcaagtta cggtaagcta tcgtatatgc aaatatgaag gaatcatggg aaataggccc
     7081 tcttcctgcc cgaccttgac gtcgactcta gaatggaggc ggtactatgt agatgagaat
     7141 tcaggagcaa actgggaaaa gcaactgctt ccaaatattt gtgattttta cagtgtagtt
     7201 ttggaaaaac tcttagccta ccaattcttc taagtgtttt aaaatgtggg agccagtaca
     7261 catgaagtta tagagtgttt taatgaggct taaatattta ccgtaactat gaaatgctac
     7321 gcatatcatg ctgttcaggc tccgtggcca cgcaactcat acttaagcag acagtggttc
     7381 aaagtttttt tcttccattt caggtgtcgt gaacaccgcc accatggaca agaagtactc
     7441 cattgggctc gatatcggta ccaacagcgt cggctgggcc gtcattacgg acgagtacaa
     7501 ggtgccgagc aaaaaattca aagttctggg caataccgat cgccacagca taaagaagaa
     7561 cctcattgga gccctcctgt tcgactccgg ggagacggcc gaagccacgc ggctcaaaag
     7621 aacagcacgg cgcagatata cccgcagaaa gaatcggatc tgctacctgc aggagatctt
     7681 tagtaatgag atggctaagg tggatgactc tttcttccat aggctggagg agtccttttt
     7741 ggtggaggag gataaaaagc acgagcgcca cccaatcttt ggcaatatcg tggacgaggt
     7801 ggcgtaccat gaaaagtacc caaccatata tcatctgagg aagaagctgg tagacagtac
     7861 tgataaggct gacttgcggt tgatctatct cgcgctggcg cacatgatca aatttcgggg
     7921 acacttcctc atcgaggggg acctgaaccc agacaacagc gatgtcgaca aactctttat
     7981 ccaactggtt cagacttaca atcagctttt cgaggagaac ccgatcaacg catccggcgt
     8041 tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc cggcggctcg aaaacctcat
     8101 cgcacagctc cctggggaga agaagaacgg cctgtttggt aatcttatcg ccctgtcact
     8161 cgggctgacc cccaacttta aatctaactt cgacctggcc gaagatgcca agctgcaact
     8221 gagcaaagac acctacgatg atgatctcga caatctgctg gcccagatcg gcgaccagta
     8281 cgcagacctt tttttggcgg caaagaacct gtcagacgcc attctgctga gtgatattct
     8341 gcgagtgaac acggagatca ccaaagctcc gctgagcgct agtatgatca agcgctatga
     8401 tgagcaccac caagacttga ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa
     8461 gtacaaggaa attttcttcg atcagtctaa aaatggctac gccggataca ttgacggcgg
     8521 agcaagccag gaggaatttt acaaatttat taagcccatc ttggaaaaaa tggacggcac
     8581 cgaggagctg ctggtaaagc tgaacagaga agatctgttg cgcaaacagc gcactttcga
     8641 caatggaagc atcccccacc agattcacct gggcgaactg cacgctatcc tcaggcggca
     8701 agaggatttc tacccctttt tgaaagataa cagggaaaag attgagaaaa tcctcacatt
     8761 tcggataccc tactatgtag gccccctcgc tcggggaaat tccagattcg cgtggatgac
     8821 tcgcaaatca gaagagacca tcactccctg gaacttcgag gaagtcgtgg ataagggggc
     8881 ctctgcccag tccttcatcg aaaggatgac taactttgat aaaaatctgc ctaacgaaaa
     8941 ggtgcttcct aaacactctc tgctgtacga gtacttcaca gtttataacg agctcaccaa
     9001 ggtcaaatac gtcacagaag ggatgagaaa gccagcattc ctgtctggag agcagaagaa
     9061 agctatcgtg gacctcctct tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga
     9121 agactatttc aaaaagattg aatgtttcga ctctgttgaa atcagcggag tggaggatcg
     9181 cttcaacgca tccctgggaa cgtatcacga tctcctgaaa atcattaaag acaaggactt
     9241 cctggacaat gaggagaacg aggacattct tgaggacatt gtcctcaccc ttacgttgtt
     9301 tgaagatagg gagatgattg aagaacgctt gaaaacttac gctcatctct tcgacgacaa
     9361 agtcatgaaa cagctcaaga gacgccgata tacaggatgg gggcggctgt caagaaaact
     9421 gatcaatggc atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga
     9481 tggatttgcc aaccggaact tcatgcagtt gatccatgat gactctctca cctttaagga
     9541 ggacatccag aaagcacaag tttctggcca gggggacagt cttcacgagc acatcgctaa
     9601 tcttgcaggt agcccagcta tcaaaaaggg aatactgcag accgttaagg tcgtggatga
     9661 actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc gttatcgaga tggcccgaga
     9721 gaaccaaact acccagaagg gacagaagaa cagtagggaa aggatgaaga ggattgaaga
     9781 gggtataaaa gaactggggt cccaaatcct taaggaacac ccagttgaaa acacccagct
     9841 tcagaatgag aagctctacc tgtactacct gcagaacggc agggacatgt acgtggatca
     9901 ggaactggac atcaaccggt tgtccgacta cgacgtggat catatcgtgc cccaaagctt
     9961 tctcaaagat gattctattg ataataaagt gttgacaaga tccgataaaa atagagggaa
    10021 gagtgataac gtcccctcag aagaagttgt caagaaaatg aaaaattatt ggcggcagct
    10081 gctgaacgcc aaactgatca cacaacggaa gttcgataat ctgactaagg ctgaacgagg
    10141 tggcctgtct gagttggata aagccggctt catcaaaagg cagcttgttg agacacgcca
    10201 gatcaccaag cacgtggccc aaattctcga ttcacgcatg aacaccaagt acgatgaaaa
    10261 tgacaaactg attcgagagg tgaaagttat tactctgaag tctaagctgg tctcagattt
    10321 cagaaaggac tttcagtttt ataaggtgag agagatcaac aattaccacc atgcgcatga
    10381 tgcctacctg aatgcagtgg taggcactgc acttatcaaa aaatatccca agctggaatc
    10441 tgaatttgtt tacggagact ataaagtgta cgatgttagg aaaatgatcg caaagtctga
    10501 gcaggaaata ggcaaggcca ccgctaagta cttcttttac agcaatatta tgaatttttt
    10561 caagaccgag attacactgg ccaatggaga gattcggaag cgaccactta tcgaaacaaa
    10621 cggagaaaca ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag tccgcaaggt
    10681 cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg gaggcttctc
    10741 caaggaaagt atcctcccga aaaggaacag cgacaagctg atcgcacgca aaaaagattg
    10801 ggaccccaag aaatacggcg gattcgattc tcctacagtc gcttacagtg tactggttgt
    10861 ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat
    10921 cacaatcatg gagcgatcca gcttcgagaa aaaccccatc gactttctcg aagcgaaagg
    10981 atataaagag gtcaaaaaag acctcatcat taagctgccc aagtactctc tctttgagct
    11041 tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct
    11101 ggcactgccc tctaaatacg ttaatttctt gtatctggcc agccactatg aaaagctcaa
    11161 agggtctccc gaagataatg agcagaagca gctgttcgtg gaacaacaca aacactacct
    11221 tgatgagatc atcgagcaaa taagcgagtt ctccaaaaga gtgatcctcg ccgacgctaa
    11281 cctcgataag gtgctttctg cttacaataa gcacagggat aagcccatca gggagcaggc
    11341 agaaaacatt atccacttgt ttactctgac caacttgggc gcgcctgcag ccttcaagta
    11401 cttcgacacc accatagaca gaaagcggta cacctctaca aaggaggtcc tggacgccac
    11461 actgattcat cagtcaatta cggggctcta tgaaacaaga atcgacctct ctcagctcgg
    11521 tggagacgag gatcccgccg ccaagcgcgt gaagctggac ggcgcaacaa acttctctct
    11581 gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atggccaagc ctttgtctca
    11641 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
    11701 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
    11761 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
    11821 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
    11881 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
    11941 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
    12001 ttatgtgtgg gagggctaag aattcgatat caagcttatc ggtaatcaac ctctggatta
    12061 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
    12121 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
    12181 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
    12241 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
    12301 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
    12361 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
    12421 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
    12481 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
    12541 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
    12601 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gagacctaga
    12661 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
    12721 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc
    12781 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
    12841 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca
    12901 aggctacttc cctgattggc agaactacac accagggcca gggatcagat atccactgac
    12961 ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga
    13021 aggagagaac acccgctt
//
