LOCUS       sequence-384497-       13013 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral Cas9/sgRNA vector targeting C-terminus of hPER1, works 
            with Addgene 189979-189982.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Gabriel CH, Del Olmo M, Rizki Widini A, Roshanbin R, Woyde J, Hamza 
            E, Gutu NN, Zehtabian A, Ewers H, Granada A, Herzel H, Kramer A
  TITLE     Circadian period is compensated for repressor protein turnover rates
            in single cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Aug 20;121(34):e2404738121. doi: 
            10.1073/pnas.2404738121. Epub 2024 Aug 14.
  PUBMED    39141353
REFERENCE   2  (bases 1 to 13013)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2024-08-20"; volume: "121(34):e2404738121. 
            doi"; pages: " 10.1073/pnas.2404738121. Epub 2024 Aug 14"
COMMENT     Sequence Label: pLentiCriprV2-sgRNA-PER1-#1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      151..286
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     213..232
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        348..395
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             414..788
                     /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    918..1039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(955..974)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1009..1028
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1088..1104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1088..1104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1101..1123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1112..1128
                     /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(1136..1142,1143..1160,1161..1166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1181..1202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1319..1336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1490..2078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1570..1589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2249..3040,3041..3109))
                     /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     2872..2891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3110..3214)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3289..3308)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3480..3859
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3861..4059
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4014..4034
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4077..4257
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4304..4429
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4922..5155
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5340..5384
                     /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    5682..5799
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5850..6098
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5850..5870
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6021..6040
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6120..6195
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6257..6468
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6487..6496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6493..10596
                     /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             6493..10596
                     /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             10597..10644
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10645..10668
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10678..10734
                     /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             10735..11331
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     11228..11248
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11347..11935
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11400..11420)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11818..11829)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12007..12240
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12266..12283)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12272..12496
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12542..12970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12629..12648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12839..12860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12979..12999)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(12984..13013,1..300)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
       61 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
      121 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
      181 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
      241 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
      301 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
      361 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
      421 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
      481 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
      541 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
      601 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
      661 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
      721 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
      781 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
      841 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
      901 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
      961 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1021 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     1081 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1141 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1201 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1261 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1321 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1381 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1441 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1501 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1561 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1621 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1681 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1741 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1801 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1861 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1921 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1981 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2041 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2101 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2161 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2221 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2281 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2341 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2401 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2461 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2521 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2581 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2641 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2701 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2761 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2821 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2881 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2941 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3001 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3061 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3121 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3181 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3241 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     3301 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     3361 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     3421 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     3481 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     3541 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     3601 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     3661 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     3721 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     3781 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     3841 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     3901 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     3961 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     4021 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt
     4081 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     4141 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     4201 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     4261 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     4321 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     4381 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     4441 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     4501 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     4561 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     4621 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     4681 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     4741 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     4801 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     4861 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     4921 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     4981 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     5041 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     5101 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     5161 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     5221 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     5281 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     5341 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     5401 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     5461 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     5521 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     5581 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
     5641 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     5701 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     5761 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     5821 agagatccag tttggttaat taaggtaccg agggcctatt tcccatgatt ccttcatatt
     5881 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
     5941 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
     6001 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
     6061 tcttggcttt atatatcttg tggaaaggac gaaacaccgg aaagcctctc atggactccg
     6121 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     6181 gcaccgagtc ggtgcttttt tgaattcgct agctaggtct tgaaaggagt gggaattggc
     6241 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     6301 ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     6361 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     6421 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagga ccggttctag
     6481 agcgctgcca ccatggacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
     6541 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     6601 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     6661 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     6721 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     6781 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     6841 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     6901 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     6961 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     7021 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     7081 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     7141 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     7201 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     7261 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     7321 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     7381 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     7441 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     7501 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     7561 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     7621 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     7681 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     7741 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     7801 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     7861 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     7921 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     7981 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     8041 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     8101 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     8161 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     8221 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     8281 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     8341 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     8401 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     8461 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     8521 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     8581 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     8641 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     8701 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     8761 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     8821 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     8881 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     8941 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     9001 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     9061 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     9121 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     9181 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     9241 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     9301 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     9361 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     9421 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     9481 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     9541 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     9601 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     9661 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     9721 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     9781 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     9841 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     9901 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     9961 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
    10021 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
    10081 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
    10141 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
    10201 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
    10261 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
    10321 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
    10381 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
    10441 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
    10501 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
    10561 gagacacgga tcgacctgtc tcagctggga ggcgacaagc gacctgccgc cacaaagaag
    10621 gctggacagg ctaagaagaa gaaagattac aaagacgatg acgataaggg atccggcgca
    10681 acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgaccgag
    10741 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
    10801 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
    10861 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
    10921 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
    10981 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
    11041 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
    11101 ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc
    11161 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
    11221 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
    11281 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa
    11341 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
    11401 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
    11461 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
    11521 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
    11581 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
    11641 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
    11701 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
    11761 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
    11821 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
    11881 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
    11941 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
    12001 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
    12061 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
    12121 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
    12181 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
    12241 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
    12301 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
    12361 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
    12421 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
    12481 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
    12541 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
    12601 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
    12661 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
    12721 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
    12781 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
    12841 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
    12901 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
    12961 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc cca
//
