LOCUS       sequence-402992-       16177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid backbone used to generate the recombination template to 
            generate the SELECTIV mice through Integrase Mediated Transgenesis. 
            Allows for Cre-dependent expression of AAVR and Cas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16177)
  AUTHORS   Zengel J, Wang YX, Seo JW, Ning K, Hamilton JN, Wu B, Raie M, 
            Holbrook C, Su S, Clements DR, Pillay S, Puschnik AS, Winslow MM, 
            Idoyaga J, Nagamine CM, Sun Y, Mahajan VB, Ferrara KW, Blau HM, 
            Carette JE
  TITLE     Hardwiring tissue-specific AAV transduction in mice through 
            engineered receptor expression.
  JOURNAL   Nat Methods. 2023 Jul;20(7):1070-1081. doi: 
            10.1038/s41592-023-01896-x. Epub 2023 Jun 8.
  PUBMED    37291262
REFERENCE   2  (bases 1 to 16177)
  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: 
            "10.1038/s41592-023-01896-x"; journalName: "Nat Methods"; date: 
            "2023-07"; volume: "20"; issue: "7"; pages: "1070-1081"
COMMENT     Sequence Label: pattB-LSL-AAVR-F2A-spCas9
FEATURES             Location/Qualifiers
     source          1..16177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(624..1415,1416..1484))
                     /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     1247..1266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1485..1589)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1615..2070)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1752..1773)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1964..1983
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2197..2219
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2211..2228
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2212..2228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2235..2254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2235..2253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2304..2373
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     enhancer        2605..2984
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2986..3263
                     /label=chicken beta-actin promoter
     intron          3264..4280
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4204..4226
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4288..4307
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     4340..4356
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4340..4356
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    4370..4403
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4433..4932
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4468..4491)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4865..4884
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4946..5749
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5009..5028)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5619..5638
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6080..6214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    6333..6467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    6586..6720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    6749..6782
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             9936..10643
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(10081..10099)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     10349..10368
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     10543..10566
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             10653..10718
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     regulatory      10734..10743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10743..10808
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             10815..10835
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10860..14960
                     /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             10860..14960
                     /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             14961..15008
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    15133..15254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    15317..15386
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(15601..15619)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(15640..15656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15640..15656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15653..15675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15664..15680
                     /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(15688..15694,15695..15712,15713..15718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15733..15754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15871..15888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(16042..16177,1..453))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16122..16141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
       61 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      121 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      181 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      241 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      301 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      361 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      421 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      481 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      541 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      601 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      721 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      781 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      841 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      901 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
      961 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1021 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1081 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1141 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1201 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1261 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1321 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1381 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1441 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1501 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1561 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     1621 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1681 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1741 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1801 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1861 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1921 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1981 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     2041 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     2101 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     2161 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     2221 cggccagtga attgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg
     2281 aggtcgacga tgtaggtcac ggtctcgaag ccgcggtgcg ggtgccaggg cgtgcccttg
     2341 ggctccccgg gcgcgtactc cacctcaccc atctggtcca tcatgatgaa cgggtcgagg
     2401 tggcggtagt tgatcccggc gaacgcgcgg cgcaccggga agccctcgcc ctcgaaaccg
     2461 ctgggcgcgg tggtcacggt gagcacggga cgtgcgacgg cgtcggcggg tgcggatacg
     2521 cggggcagcg tcagcgggtt ctcgacggtc acggcgggca tgtcgacggt atcgataagc
     2581 ttcccttaat taacctcgag attcgacatt gattattgac tagttattaa tagtaatcaa
     2641 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2701 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2761 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2821 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2881 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2941 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     3001 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     3061 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccrggsg
     3121 gggsggggsg gggsgrgggg sggggsgggg cgaggcggag aggtgcggcg gcagccaatc
     3181 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     3241 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
     3301 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
     3361 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
     3421 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
     3481 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
     3541 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     3601 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     3661 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     3721 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     3781 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     3841 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggrgggctcg ggggaggggc
     3901 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     3961 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     4021 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     4081 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     4141 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     4201 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     4261 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     4321 ttggcaaaga attggccgct ctagaactag tggatccgga acccttaata taacttcgta
     4381 taatgtatgc tatacgaagt tattaggtcc ctcgacctgc aggaattcta ccgggtaggg
     4441 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     4501 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     4561 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     4621 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     4681 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     4741 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     4801 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     4861 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     4921 ccgggccttt cgacctgcag ccaatatggg atcggccatt gaacaagatg gattgcacgc
     4981 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5041 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5101 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5161 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5221 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5281 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5341 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5401 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5461 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg tgacccatgg
     5521 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5581 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5641 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5701 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag gggatccgct
     5761 gtaagtctgc agaaattgat gatctattaa acaataaaga tgtccactaa aatggaagtt
     5821 tttcctgtca tactttgtta agaagggtga gaacagagta cctacatttt gaatggaagg
     5881 attggagcta cgggggtggg ggtggggtgg gattagataa atgcctgctc tttactgaag
     5941 gctctttact attgctttat gataatgttt catagttgga tatccttata ccacatttgt
     6001 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     6061 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     6121 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6181 caaactcatc aatgtatctt atcatgtctg gatctgcgac tctgaggatc ataatcagcc
     6241 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     6301 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     6361 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6421 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctgc gactctgagg
     6481 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     6541 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     6601 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6661 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     6721 cccatcaagc tgatccggaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     6781 attagggcac catggagaag agactgggag tcaagccaag tcccgcttcc tgggttttgc
     6841 caggatattg ttggcagaca tcagtgaagc tgccgagaag cctgtacctg ctttacagtt
     6901 tcttctgctt cagcgttctg tggttgtcaa cagatgctga tgagagcaga tgccaacagg
     6961 ggaagacact ttatggagct ggcttgagaa ctgagggaga aaatcacctc cggcttcttg
     7021 caggaagcct gcctttccac gcctgtcggg ctgcctgctg ccgggactct gcctgccacg
     7081 ctctatggtg gctggaaggg atgtgctttc aggctgactg cagcaagccc cagagctgcc
     7141 agccttttag gacagactct tccaattcca tgctgatcat ttttcaaaaa tcccaaacta
     7201 cagatgattt gggccttctg cctgaagatg atgaaccaca tcttctgagg ctaggctggg
     7261 gcaggacatc gtggaggagg cagagccttc ttggggctcc cctcaccctt tctgtaccct
     7321 ctagtcacca ccagagctta ctcagggatc ggcagaagag agatctcagt gtggtaccta
     7381 cacatggagc gatgcagcat tctaaagtga atcactccga ggaagcaggt gctctgagtc
     7441 ccacctctgc agaggtccgc aaaaccatta cagttgccgg ttccttcacc agtaaccaca
     7501 ctacacagac tcctgagtgg cccaagaatg tgtccatcca tcctgaacca tccgagcact
     7561 ccagtcctgt atctggtact ccgcaagtaa aaagcactga gcacagtcca actgatgccc
     7621 ctctgccagt ggccccctcc tacagctatg ccacccccac gccccaggcc tcttctcaga
     7681 gcacctcagc accacaccca gttgtaaagg agctggtggt gtctgctggg aagagcgtcc
     7741 agatcaccct gcctaagaat gaagttcagt taaatgcctt cgtccttcca gaagcagagc
     7801 caggagaaac ctacacctac gactggcagc tgatcactca tcctacagac tacagtggag
     7861 aggtggagag gaaacattcc cagagcctcc aactgtccaa gctgactcca ggcctgtacg
     7921 aattcaaggt gactgtggat ggccagaatg cccatgggga aggctacgtg aatgtgacag
     7981 tgaaaccaga gccccgtaag aaccggcctc ccgttgctgt ggtgtcacct cagttccagg
     8041 agatctcgct gccaaccact tctaccatca ttgatggcag ccagagcacg gatgacgata
     8101 aaattgtcca gtaccactgg gaagagctta aggggcccct gagagaagag aagatctctg
     8161 aagacacagc catactaaaa cttagtaagc tcgtcccggg gaactacacc ttcagcttaa
     8221 ctgttgtcga ctctgacggg gctaccaact ccaccactgc aagcctgact gtgaacaaag
     8281 ctgtggacta ccctcccgtg gccaatgcag gccccaacca agtgatcacc ctgcctcaga
     8341 actccatcac cctctttgga aaccagagca cggatgacca cggcatcacc agctatgagt
     8401 ggtcgctcag cccgagcagc aaagggaagg tggtggagat gcagggagtt agaacgccag
     8461 ccctgcagct gtccgcaatg caagaaggag actataccta ccagctcaca gtgactgaca
     8521 ccgcaggaca acaggccacc gcccaagtga ctgtgattgt gcagcctgag aacaacaagc
     8581 ctcctcaggc agatgcaggc ccagacaaag agctgaccct gcccgtggac agcacaaccc
     8641 tggacggcag caagagcaca gatgaccaga gagtcgtctc ttacctttgg gagcagagtc
     8701 ggggacctga cggggtgcag ctggagaatg ccaacagcag tgtcgccact gtgactgggc
     8761 tgcaagtcgg gacttatgta ttcaccttga ctgtcaaaga tgagaggaac ctacagagcc
     8821 agagctccgt taatgtcatt gtcaaagaag aaataaacaa accgccagta gccaagatcg
     8881 ctgggaacgt ggtggtgacc ttgcccacga gcacagctga gctggatggc tcgaggtcct
     8941 cagatgacaa ggggatagtc agctacctgt ggactcgaga tgagacgagc ccagccgcag
     9001 gggaggtgct gaatcactct gaccaccacc ctgtcctctt cctctccaac ctggtggagg
     9061 ggacctacac gtttcacctg aaagtgacag atgcaaaggg cgagagcgac acagaccgga
     9121 cgacagtgga agtgaagcct gaccccagga aaagcaacct agtggagatc atcttggatg
     9181 tgaacgtcag tcagctgact gagaggctga aggggatgct catccgccag attggggtcc
     9241 tcctgggggt gctggattcc gacatcattg tgcaaaagat tcagccgtac acggagcaga
     9301 gcaccaagat gttgtttttt gttcagaacg accctcccca ccagctcttc aaaggccatg
     9361 aggtggcagc catgctcaag agcgagctgc agaagcagaa ggctgacttc ctcatcttca
     9421 gagccctgga aatcagcaca gtcacatgtc agctgaactg ttctgaccat ggccactgtg
     9481 actcattcac caagcgctgt gtctgtgacc cgttttggat ggagaatttc atcaaggtgc
     9541 agctgaggga tggagacagc aactgtgaat ggagcgtgct ctacgtcatc attgcctcct
     9601 ttgtcattgt tgttgccttg gggatcctgt catggactac aatctgctgc tgcaagaggc
     9661 aaaaaggaaa acccaagagg aaaagcagat acaagatcct ggatgccaca gatcaggaga
     9721 gcctggagct gaaaccaacc tcccgagcag gcagcaaaca gaaaggcccc acgctgagca
     9781 gcagcctgat gcattctgaa tcggagctgg acagcgacga tgccatcttc acatggccag
     9841 accgggagaa gggcaaacta ctgtatggtc agaatggctc tgtgccaaac gggcaaacac
     9901 ctttgaagtc caggagcgca cgggaggaga tcttgatggt gagcaagggc gaggaggata
     9961 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
    10021 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
    10081 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
    10141 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
    10201 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
    10261 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
    10321 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
    10381 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
    10441 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacaaggcca
    10501 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagctggac atcacctccc
    10561 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg
    10621 gcggcatgga cgagctgtac aagggaagcg gagtgaaaca gactttgaat tttgaccttc
    10681 tcaagttggc gggagacgtg gagtccaacc ctggacctcc ccaaacgacc ggtgccacca
    10741 tggactataa ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg
    10801 acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg
    10861 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
    10921 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
    10981 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
    11041 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
    11101 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
    11161 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
    11221 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
    11281 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
    11341 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
    11401 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
    11461 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
    11521 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
    11581 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
    11641 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
    11701 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
    11761 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
    11821 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
    11881 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
    11941 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
    12001 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
    12061 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
    12121 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
    12181 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
    12241 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
    12301 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
    12361 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
    12421 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
    12481 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
    12541 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
    12601 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
    12661 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
    12721 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
    12781 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
    12841 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
    12901 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
    12961 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
    13021 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
    13081 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
    13141 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
    13201 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
    13261 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
    13321 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg
    13381 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
    13441 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
    13501 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
    13561 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
    13621 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
    13681 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
    13741 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
    13801 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
    13861 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
    13921 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
    13981 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
    14041 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
    14101 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
    14161 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
    14221 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
    14281 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
    14341 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
    14401 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
    14461 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
    14521 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
    14581 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
    14641 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
    14701 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
    14761 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
    14821 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
    14881 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
    14941 tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
    15001 agaaaaagta agaattctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
    15061 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
    15121 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    15181 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    15241 atcaatgtat cttatttaaa ttgggcgcgc ccaactagtt ctagagcggc cgccactcga
    15301 cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca gggcgtgccc ttgggctccc
    15361 cgggcgcgta ctccacctca cccatctggt ccatcatgat gaacgggtcg aggtggcggt
    15421 agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc gccctcgaaa ccgctgggcg
    15481 cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc gggtgcggat acgcggggca
    15541 gcgtcagcgg gttctcgacg gtcacggcgg gcatgtcgag tggagctcca gcttttgttc
    15601 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
    15661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
    15721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
    15781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
    15841 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
    15901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
    15961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
    16021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
    16081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
    16141 ccctggaagc tccctcgtgc gctctcctgt tccgacc
//
