LOCUS       sequence-417650-       14207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  codes for 2 gRNA-TracrRNA targeting human OCLN, Cas9, mNeonGreen 
            fluorescent protein and puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14207)
  AUTHORS   Baptiste Pradel , Maika S. Deffieu, Veronique Robert-Hebmann, 
            Guilhem Cantaloube, Mathias Faure, Nathalie Chazal, View ORCID 
            ProfileRaphael Gaudin, View ORCID ProfileLucile Espert
  TITLE     LC3B conjugation machinery promotes autophagy-independent HIV-1 
            entry in CD4+ T lymphocytes
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 14207)
  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: "bioRxiv"
COMMENT     Sequence Label: pLenti gRNA KO OCLNx2 spCas9-T2A-mNeonGreen-P2...
FEATURES             Location/Qualifiers
     source          1..14207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(34..282)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(92..111)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(262..282)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    346..463
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(761..805)
                     /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    990..1223
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1716..1841
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1888..2068
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2086..2284)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2111..2131)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2286..2665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2837..2856
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2931..3035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3036..3104,3105..3896)
                     /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     complement(3254..3273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4067..4655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4556..4575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4809..4826
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4943..4964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4979..4984,4985..5002,5003..5009)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5017..5033
                     /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)."
     primer_bind     5022..5044
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5041..5057
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5041..5057
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5106..5227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5117..5136)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5171..5190
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5357..5731)
                     /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"
     promoter        complement(5750..5797)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5845..6174)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5859..5994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5913..5932)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6159..6179
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6188..6616)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6298..6319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6510..6529
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6662..6886
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6875..6892
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6918..7151
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    7223..7811
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7329..7340
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7738..7758
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7827..8423)
                     /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     complement(7910..7930)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             complement(8424..8480)
                     /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             complement(8508..9212)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      8951..8960
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(9213..9266)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(9273..9320)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(9321..13421)
                     /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             complement(9321..13421)
                     /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             complement(13446..13466)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(13473..13538)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      13538..13547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(13555..13766)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(13827..13902)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(13923..14137)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(13983..14002)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        complement(join(14151..14207,1..19))
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ctatttctag ctctaaaaca ggtggacttt caagaggccc cgtcctttcc acaagatata
       61 taaagccaag aaatcgaaat actttcaagt tacggtaagc atatgatagt ccattttaaa
      121 acataatttt aaaactgcaa actacccaag aaattattac tttctacgtc acgtattttg
      181 tactaatatc tttgtgttta cagtcaaatt aattccaatt atctctctaa cagccttgta
      241 tcgtatatgc aaatatgaag gaatcatggg aaataggccc tcttcctgcc cgacctttat
      301 cttaattaac caaactggat ctctgctgtc cctgtaataa acccgaaaat tttgaatttt
      361 tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct
      421 ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc
      481 catttgtctg cagaattggc gcacgcagtg ccgatccgtt cactaatcga atggatctgt
      541 ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt cccctcgggg
      601 ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag
      661 aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt
      721 ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta
      781 attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt
      841 gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc
      901 aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa
      961 tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca
     1021 cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct
     1081 gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt
     1141 gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca
     1201 agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc
     1261 tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata
     1321 attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc agcggccgct
     1381 tgctgtgcgg tggtcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg
     1441 gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat
     1501 gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta
     1561 cagccttctg atgtttctaa caggccagga ttaactgcga atcgttctag ctccctgctt
     1621 gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt
     1681 tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc
     1741 ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt
     1801 gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc
     1861 gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc
     1921 tgagggatct ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc
     1981 aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg
     2041 ctcagatctg gtctaaccag agagacccag tacaggcaaa acgcgctgct tatatagacc
     2101 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     2161 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
     2221 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
     2281 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     2341 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     2401 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     2461 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     2521 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     2581 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     2641 taataactag tcaataatca atgtcaacgc gtatatctgg cccgtacatc gcgaagcagc
     2701 gcaaaacgcc taaccctaag cagattcttc atgcaattgt cggtcaagcc ttgccttgtt
     2761 gtagcttaaa ttttgctcgc gcactactca gcgacctcca acacacaagc agggagcaga
     2821 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca taggggatcg
     2881 ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2941 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3001 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3061 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3121 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3181 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3241 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3301 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3361 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3421 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3481 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3541 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3601 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3661 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3721 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3781 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3841 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3901 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3961 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4081 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4141 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4201 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4261 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4321 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4381 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4441 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4501 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4561 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4621 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4681 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4741 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4801 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4861 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4921 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4981 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5041 caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
     5101 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     5161 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     5221 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     5281 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     5341 aatctcgtag cacgtgtcag tcctgctcct cggccacgaa gtgcacgcag ttgccggccg
     5401 ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc gatctcggtc atggccggcc
     5461 cggaggcgtc ccggaagttc gtggacacga cctccgacca ctcggcgtac agctcgtcca
     5521 ggccgcgcac ccacacccag gccagggtgt tgtccggcac cacctggtcc tggaccgcgc
     5581 tgatgaacag ggtcacgtcg tcccggacca caccggcgaa gtcgtcctcc acgaagtccc
     5641 gggagaaccc gagccggtcg gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga
     5701 gcaccggaac ggcactggtc aacttggcca tggtttagtt cctcaccttg tcgtattata
     5761 ctatgccgat atactatgcc gatgattaat tgtcaacacg tgctgatcag atccgaaaat
     5821 ggatatacaa gctcccggga gctttttgca aaagcctagg cctccaaaaa agcctcctca
     5881 ctacttctgg aatagctcag aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat
     5941 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     6001 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     6061 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     6121 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     6181 cagaattaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     6241 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     6301 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     6361 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     6421 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     6481 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6541 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     6601 ccgctacagg gcgcgtgggg atacccccta gagccccagc tggttctttc cgcctcagaa
     6661 gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg
     6721 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
     6781 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
     6841 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag
     6901 cgggtttaaa cgggccctgc tagagatttt ccacactgac taaaagggtc tgagggatct
     6961 ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct
     7021 ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg
     7081 gtctaaccag agagacccag tacaagcaaa aagcagatct tgtcttcgtt gggagtgaat
     7141 tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta
     7201 agtcattggt cttaaagtcg acgcggggag gcggcccaaa gggagatccg actcgtctga
     7261 gggcgaaggc gaagacgcgg aagaggccgc agagccggca gcaggccgcg ggaaggaagg
     7321 tccgctggat tgagggccga agggacgtag cagaaggacg tcccgcgcag aatccaggtg
     7381 gcaacacagg cgagcagcca aggaaaggac gatgatttcc ccgacaacac cacggaattg
     7441 tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc
     7501 gccgtggcaa tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca
     7561 atgccccaac cagtgggggt tgcgtcagca aacacagtgc acaccacgcc acgttgcctg
     7621 acaacgggcc acaactcctc ataaagagac agcaaccagg atttatacaa ggaggagaaa
     7681 atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat
     7741 agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc
     7801 agaggttgat tgtcgactta acgcgttcag gcaccgggct tgcgggtcat gcaccaggtg
     7861 cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag
     7921 gggaggttgc ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg ctcggccgcc
     7981 tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagact
     8041 ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc
     8101 atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg
     8161 gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg
     8221 tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc
     8281 agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag
     8341 tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg
     8401 cgcaccgtgg gcttgtactc ggtcggtcca ggattctctt cgacatctcc ggcttgtttc
     8461 agcagagaga agtttgttgc gccggatccg gtaccagcgc ttctagactt gtacagctcg
     8521 tccatgccca tcacatcggt aaaggccttt tgccactcct tgaagttgag ctcggtcttg
     8581 gagtgcttga gctccgtctt acggaacacg tacatcggct ggttcttcag atagttagcc
     8641 gccattggct tggcaaaggt gtaggtggtc cgcgcagtgc tccggtagcg cttgccattt
     8701 ccagtggtgt aactccactt aaaggtactg atgatggttt tgtcgttggg gtaagtcttc
     8761 ttcgacctgc accagtccgc agcggtcagc gagttggtca tcacaggacc gtcagcaggg
     8821 aaaccagtcc ccttcacctg ggcctctcct ttgatgtggc ttccctcgta ggtgtagcgg
     8881 tagttaacag taagggaggc accatcttca aactgcattg tgcgatggac ttggtatccg
     8941 gagccatcta ccatggcggc ctggaaaggc gacatcccgt cagggtaggg caggtactga
     9001 tggaagccat acccgatatg agggaccaga atccaggggg agaactggag gtcacccttg
     9061 gtggacttca ggtttaactc ctcataacca tcatttggat tgccggtgcc ctgacccacc
     9121 atgtcaaagt ccacaccgtt gatggagcca aagatgtgta actcatgtgt cgctgggaga
     9181 gaggccatgt tatcctcctc gcccttgctc actgggccag gattctcctc gacgtcaccg
     9241 catgttagca gacttcctct gccctcgaat tcctttttct tttttgcctg gccggccttt
     9301 ttcgtggccg ccggcctttt gtcgcctccc agctgagaca ggtcgatccg tgtctcgtac
     9361 aggccggtga tgctctggtg gatcagggtg gcgtccagca cctctttggt gctggtgtac
     9421 ctcttccggt cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc cagattggtc
     9481 agggtaaaca ggtggatgat attctcggcc tgctctctga tgggcttatc ccggtgcttg
     9541 ttgtaggcgg acagcacttt gtccagatta gcgtcggcca ggatcactct cttggagaac
     9601 tcgctgatct gctcgatgat ctcgtccagg tagtgcttgt gctgttccac aaacagctgt
     9661 ttctgctcat tatcctcggg ggagcccttc agcttctcat agtggctggc caggtacagg
     9721 aagttcacat atttggaggg cagggccagt tcgtttccct tctgcagttc gccggcagag
     9781 gccagcattc tcttccggcc gttttccagc tcgaacaggg agtacttagg cagcttgatg
     9841 atcaggtcct ttttcacttc tttgtagccc ttggcttcca gaaagtcgat gggattcttc
     9901 tcgaagctgc ttctttccat gatggtgatc cccagcagct ctttcacact cttcagtttc
     9961 ttggacttgc ccttttccac tttggccacc accagcacag aataggccac ggtggggctg
    10021 tcgaagccgc cgtacttctt agggtcccag tccttctttc tggcgatcag cttatcgctg
    10081 ttcctcttgg gcaggataga ctctttgctg aagccgcctg tctgcacctc ggtctttttc
    10141 acgatattca cttggggcat gctcagcact ttccgcacgg tggcaaaatc ccggccctta
    10201 tcccacacga tctccccggt ttcgccgttt gtctcgatca gaggccgctt ccggatctcg
    10261 ccgttggcca gggtaatctc ggtcttgaaa aagttcatga tgttgctgta gaagaagtac
    10321 ttggcggtag ccttgccgat ttcctgctcg ctcttggcga tcatcttccg cacgtcgtac
    10381 accttgtagt cgccgtacac gaactcgctt tccagcttag ggtacttttt gatcagggcg
    10441 gttcccacga cggcgttcag gtaggcgtcg tgggcgtggt ggtagttgtt gatctcgcgc
    10501 actttgtaaa actggaaatc cttccggaaa tcggacacca gcttggactt cagggtgatc
    10561 actttcactt cccggatcag cttgtcattc tcgtcgtact tagtgttcat ccgggagtcc
    10621 aggatctgtg ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct cttgatgaag
    10681 ccggccttat ccagttcgct caggccgcct ctctcggcct tggtcagatt gtcgaacttt
    10741 ctctgggtaa tcagcttggc gttcagcagc tgccgccagt agttcttcat cttcttcacg
    10801 acctcttcgg agggcacgtt gtcgctcttg ccccggttct tgtcgcttct ggtcagcacc
    10861 ttgttgtcga tggagtcgtc cttcagaaag ctctgaggca cgatatggtc cacatcgtag
    10921 tcggacagcc ggttgatgtc cagttcctgg tccacgtaca tatcccgccc attctgcagg
    10981 tagtacaggt acagcttctc gttctgcagc tgggtgtttt ccacggggtg ttctttcagg
    11041 atctggctgc ccagctcttt gatgccctct tcgatccgct tcattctctc gcggctgttc
    11101 ttctgtccct tctgggtggt ctggttctct ctggccattt cgatcacgat gttctcgggc
    11161 ttgtgccggc ccatcacttt cacgagctcg tccaccacct tcactgtctg caggatgccc
    11221 ttcttaatgg cggggctgcc ggccagattg gcaatgtgct cgtgcaggct atcgccctgg
    11281 ccggacacct gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc gtggatcagc
    11341 tgcatgaagt ttctgttggc gaagccgtcg gacttcagga aatccaggat tgtcttgccg
    11401 gactgcttgt cccggatgcc gttgatcagc ttccggctca gcctgcccca gccggtgtat
    11461 ctccgccgct tcagctgctt catcactttg tcgtcgaaca ggtgggcata ggttttcagc
    11521 cgttcctcga tcatctctct gtcctcaaac agtgtcaggg tcagcacgat atcttccaga
    11581 atgtcctcgt tttcctcatt gtccaggaag tccttgtcct tgataatttt cagcagatcg
    11641 tggtatgtgc ccagggaggc gttgaaccga tcttccacgc cggagatttc cacggagtcg
    11701 aagcactcga ttttcttgaa gtagtcctct ttcagctgct tcacggtcac tttccggttg
    11761 gtcttgaaca gcaggtccac gatggccttt ttctgctcgc cgctcaggaa ggcgggcttt
    11821 ctcattccct cggtcacgta tttcactttg gtcagctcgt tatacacggt gaagtactcg
    11881 tacagcaggc tgtgcttggg cagcaccttc tcgttgggca ggttcttatc gaagttggtc
    11941 atccgctcga tgaagctctg ggcggaagcg cccttgtcca ccacttcctc gaagttccag
    12001 ggggtgatgg tttcctcgct ctttctggtc atccaggcga atctgctgtt tcccctggcc
    12061 agagggccca cgtagtaggg gatgcggaag gtcaggatct tctcgatctt ttcccggttg
    12121 tccttcagga atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag ctctcccagg
    12181 tggatctggt gggggatgct gccgttgtcg aaggtccgct gcttccgcag caggtcctct
    12241 ctgttcagct tcacgagcag ttcctcggtg ccgtccatct tttccaggat gggcttgatg
    12301 aacttgtaga actcttcctg gctggctccg ccgtcaatgt agccggcgta gccgttcttg
    12361 ctctggtcga agaaaatctc tttgtacttc tcaggcagct gctgccgcac gagagctttc
    12421 agcagggtca ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc gctcaggggg
    12481 gccttggtga tctcggtgtt cactctcagg atgtcgctca gcaggatggc gtcggacagg
    12541 ttcttggcgg ccagaaacag gtcggcgtac tggtcgccga tctgggccag caggttgtcc
    12601 aggtcgtcgt cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc caggtcgaag
    12661 ttgctcttga agttgggggt caggcccagg ctcagggcaa tcaggtttcc gaacaggcca
    12721 ttcttcttct cgccgggcag ctgggcgatc agattttcca gccgtctgct cttgctcagt
    12781 ctggcagaca ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt ttcctcgaac
    12841 agctggttgt aggtctgcac cagctggatg aacagcttgt ccacgtcgct gttgtcgggg
    12901 ttcaggtcgc cctcgatcag gaagtggccc cggaacttga tcatgtgggc cagggccaga
    12961 tagatcagcc gcaggtcggc cttgtcggtg ctgtccacca gtttctttct caggtggtag
    13021 atggtggggt acttctcgtg gtaggccacc tcgtccacga tgttgccgaa gatggggtgc
    13081 cgctcgtgct tcttatcctc ttccaccagg aaggactctt ccagtctgtg gaagaagctg
    13141 tcgtccacct tggccatctc gttgctgaag atctcttgca gatagcagat ccggttcttc
    13201 cgtctggtgt atcttcttct ggcggttctc ttcagccggg tggcctcggc tgtttcgccg
    13261 ctgtcgaaca gcagggctcc gatcaggttc ttcttgatgc tgtgccggtc ggtgttgccc
    13321 agcaccttga atttcttgct gggcaccttg tactcgtcgg tgatcacggc ccagcccaca
    13381 gagttggtgc cgatgtccag gccgatgctg tacttcttgt cggctgctgg gactccgtgg
    13441 ataccgacct tccgcttctt ctttggggcc atcttatcgt catcgtcttt gtaatcaata
    13501 tcatgatcct tgtagtctcc gtcgtggtcc ttatagtcca tggtggcacc ggtcctgtgt
    13561 tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg
    13621 ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac
    13681 cccgcgccac cttctctagg caccggatca attgccgacc cctcccccca acttctcggg
    13741 gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagccaattc ccactccttt
    13801 caagacctag ctagattgta aaaaaagcac cgactcggtg ccactttttc aagttgataa
    13861 cggactagcc ttattttaac ttgctatttc tagctctaaa acttcaagag gcctggatga
    13921 cagtggtctc atacagaact tataagattc ccaaatccaa agacatttca cgtttatggt
    13981 gatttcccag aacacatagc gacatgcaaa tattgcaggg cgccactccc ctgtccctca
    14041 cagccatctt cctgccaggg cgcacgcgcg ctgggtgttc ccgcctagtg acactgggcc
    14101 cgcgattcct tggagcgggt tgatgacgtc agcgttcctc gagaaaaaaa gcaccgactc
    14161 ggtgccactt tttcaagttg ataacggact agccttattt taacttg
//
