LOCUS       sequence-384498-       13013 bp    DNA     circular SYN 15-DEC-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 Dec 15, 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-#2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..30
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(149..169)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        154..483
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      334..469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     396..415
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        531..578
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             597..971
                     /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    1101..1222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1138..1157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1192..1211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1271..1287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1271..1287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1284..1306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1295..1311
                     /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(1319..1325,1326..1343,1344..1349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1364..1385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1502..1519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1673..2261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1753..1772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2432..3223,3224..3292))
                     /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     3055..3074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3293..3397)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3472..3491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3663..4042
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4044..4242
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4197..4217
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4260..4440
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4487..4612
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5105..5338
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5523..5567
                     /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    5865..5982
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6033..6281
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6033..6053
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6204..6223
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6303..6378
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6440..6651
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6670..6679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6676..10779
                     /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             6676..10779
                     /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             10780..10827
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10828..10851
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10861..10917
                     /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             10918..11514
                     /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     11411..11431
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11530..12118
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11583..11603)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12001..12012)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12190..12423
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12449..12466)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12455..12679
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12725..13013,1..140)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12812..12831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
       61 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      121 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      301 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      421 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      481 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
      541 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
      601 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
      661 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
      721 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
      781 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
      841 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
      901 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
      961 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     1021 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     1081 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     1141 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1201 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     1261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1321 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1861 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2101 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3421 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     3481 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     3541 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     3601 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     3661 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     3721 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     3781 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     3841 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     3901 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     3961 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     4021 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     4081 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     4141 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     4201 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg
     4261 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     4321 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     4381 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     4441 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     4501 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     4561 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     4621 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     4681 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     4741 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     4801 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     4861 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     4921 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     4981 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     5041 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     5101 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     5161 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     5221 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     5281 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     5341 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     5401 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     5461 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     5521 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     5581 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     5641 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     5701 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     5761 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     5821 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     5881 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     5941 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     6001 agcagagatc cagtttggtt aattaaggta ccgagggcct atttcccatg attccttcat
     6061 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
     6121 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     6181 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     6241 atttcttggc tttatatatc ttgtggaaag gacgaaacac cgtaggagaa gaaagcctct
     6301 cagttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     6361 gtggcaccga gtcggtgctt ttttgaattc gctagctagg tcttgaaagg agtgggaatt
     6421 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     6481 ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     6541 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     6601 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggttc
     6661 tagagcgctg ccaccatgga caagaagtac agcatcggcc tggacatcgg caccaactct
     6721 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     6781 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
     6841 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     6901 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     6961 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     7021 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     7081 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     7141 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     7201 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     7261 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     7321 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     7381 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     7441 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     7501 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     7561 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     7621 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     7681 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     7741 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     7801 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     7861 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     7921 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     7981 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     8041 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     8101 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     8161 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     8221 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     8281 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     8341 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     8401 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     8461 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     8521 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     8581 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     8641 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     8701 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     8761 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     8821 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     8881 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     8941 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     9001 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     9061 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     9121 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     9181 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     9241 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     9301 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     9361 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     9421 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     9481 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     9541 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     9601 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     9661 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     9721 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     9781 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     9841 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     9901 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     9961 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
    10021 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
    10081 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
    10141 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
    10201 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
    10261 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
    10321 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
    10381 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
    10441 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
    10501 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
    10561 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
    10621 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
    10681 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
    10741 tacgagacac ggatcgacct gtctcagctg ggaggcgaca agcgacctgc cgccacaaag
    10801 aaggctggac aggctaagaa gaagaaagat tacaaagacg atgacgataa gggatccggc
    10861 gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgacc
    10921 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
    10981 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
    11041 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
    11101 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
    11161 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
    11221 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
    11281 tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct gggcagcgcc
    11341 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
    11401 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
    11461 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaacgcgt
    11521 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
    11581 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
    11641 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
    11701 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
    11761 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
    11821 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
    11881 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
    11941 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
    12001 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
    12061 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
    12121 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
    12181 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
    12241 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
    12301 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
    12361 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
    12421 gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
    12481 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
    12541 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
    12601 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
    12661 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
    12721 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
    12781 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
    12841 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
    12901 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
    12961 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gtt
//
