LOCUS       sequence-355783-       12612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human synapsin (hSyn)-driven spCas9 lentiviral vector for 
            CRISPR/HITI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12612)
  AUTHORS   Vevea JD, Kusick GF, Courtney KC, Chen E, Watanabe S, Chapman ER
  TITLE     Synaptotagmin 7 is targeted to the axonal plasma membrane through 
            gamma-secretase processing to promote synaptic vesicle docking in 
            mouse hippocampal neurons.
  JOURNAL   Elife. 2021 Sep 20;10. pii: 67261. doi: 10.7554/eLife.67261.
  PUBMED    34543184
REFERENCE   2  (bases 1 to 12612)
  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: "Elife."; 
            date: "2021-09-20"; volume: "10. pii: 67261. doi"; pages: " 
            10.7554/eLife.67261"
COMMENT     Sequence Label: pF(UG) hSyn spCas9
FEATURES             Location/Qualifiers
     source          1..12612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        86..415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      266..401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     328..347
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        463..510
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             529..903
                     /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    1033..1154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1070..1089)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1124..1143
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1203..1219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1203..1219)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1216..1238)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1227..1243
                     /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(1251..1257,1258..1275,1276..1281))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1434..1451)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1605..2193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1685..1704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2364..3155,3156..3224))
                     /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     2987..3006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3225..3329)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3404..3423)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3595..3974
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3976..4174
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4129..4149
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4192..4372
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4419..4544
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5037..5270
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5455..5499
                     /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    5797..5914
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5959..6405
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             6434..6460
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6464..6484
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6494..10594
                     /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             6494..10594
                     /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             10598..10618
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    10661..11249
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10714..10734)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11132..11143)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11252..11268)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11253..11269)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11774..11954
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11980..11997)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11986..12210
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12256..12612,1..72)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12343..12362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12553..12574
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
       61 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      241 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
      361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      421 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
      481 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
      541 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
      601 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
      661 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
      721 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
      781 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
      841 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
      901 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
      961 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     1021 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1081 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1141 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     1201 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1261 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1321 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1381 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1441 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1501 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1561 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1621 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1681 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1741 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1801 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1861 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1921 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1981 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2041 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2101 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2161 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2221 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2281 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2341 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2401 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2461 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2521 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2581 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2641 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2701 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2761 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2821 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2881 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2941 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3001 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3061 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3121 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3181 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3241 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3301 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3361 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     3421 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     3481 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     3541 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     3601 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     3661 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     3721 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3781 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     3841 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     3901 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     3961 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4021 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4081 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4141 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
     4201 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4261 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4321 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     4381 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     4441 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     4501 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     4561 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     4621 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     4681 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     4741 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     4801 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     4861 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     4921 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     4981 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     5041 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     5101 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     5161 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     5221 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     5281 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     5341 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     5401 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     5461 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     5521 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     5581 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     5641 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     5701 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     5761 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     5821 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     5881 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     5941 tccagtttgg ttaattaaag tgcaagtggt tttaggacca ggatgaggcg gggtgggggt
     6001 gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt
     6061 gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg
     6121 ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc
     6181 ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgtaaactcc ccttcccggc
     6241 caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag
     6301 ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag
     6361 tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcaggtacg gctagcggat
     6421 cggtgccacc atgtacccat acgatgttcc agattacgct tcgccgaaga aaaagcgcaa
     6481 ggtcgaagcg tccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     6541 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     6601 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     6661 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     6721 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     6781 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     6841 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     6901 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     6961 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     7021 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     7081 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     7141 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     7201 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     7261 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     7321 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     7381 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     7441 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     7501 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     7561 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     7621 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     7681 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     7741 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     7801 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     7861 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     7921 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     7981 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     8041 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     8101 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     8161 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     8221 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     8281 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     8341 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     8401 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     8461 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     8521 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     8581 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     8641 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     8701 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     8761 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     8821 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     8881 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     8941 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     9001 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     9061 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     9121 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     9181 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     9241 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     9301 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     9361 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     9421 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     9481 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     9541 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     9601 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     9661 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     9721 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     9781 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     9841 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     9901 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     9961 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
    10021 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
    10081 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
    10141 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
    10201 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
    10261 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
    10321 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
    10381 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
    10441 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
    10501 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
    10561 gacacggatc gacctgtctc agctgggagg cgacagcccc aagaagaaga gaaaggtgga
    10621 ggccagctaa gaactagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
    10681 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
    10741 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
    10801 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
    10861 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
    10921 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
    10981 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
    11041 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
    11101 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
    11161 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
    11221 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa
    11281 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
    11341 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
    11401 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
    11461 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
    11521 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
    11581 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
    11641 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
    11701 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
    11761 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
    11821 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
    11881 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
    11941 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
    12001 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
    12061 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
    12121 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
    12181 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
    12241 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
    12301 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
    12361 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
    12421 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
    12481 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
    12541 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
    12601 attcttttga tt
//
