LOCUS       sequence-426680-       13013 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector expressing Cas9 and an sgRNA targeting human 
            PQBP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Scott TM, Arnold LM, Powers JA, McCann DA, Rowe AB, Christensen DE, 
            Pereira MJ, Zhou W, Torrez RM, Iwasa JH, Kranzusch PJ, Sundquist WI,
            Johnson JS
  TITLE     Cell-free assays reveal that the HIV-1 capsid protects reverse 
            transcripts from cGAS immune sensing.
  JOURNAL   PLoS Pathog. 2025 Jan 28;21(1):e1012206. doi: 
            10.1371/journal.ppat.1012206.
  PUBMED    39874383
REFERENCE   2  (bases 1 to 13013)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2025-01-28"; volume: "21(1):e1012206. doi"; pages: 
            " 10.1371/journal.ppat.1012206"
COMMENT     Sequence Label: LCV2 PQBP1 KO2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        135..182
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             201..575
                     /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    705..826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(742..761)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     796..815
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(875..891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(875..891)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(888..910)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    899..915
                     /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(923..929,930..947,948..953))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    968..989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1106..1123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1277..1865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1357..1376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2036..2827,2828..2896))
                     /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     2659..2678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2897..3001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3076..3095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3267..3646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3648..3846
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3801..3821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3864..4044
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4091..4216
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4709..4942
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5127..5171
                     /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    5469..5586
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5637..5885
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5637..5657
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5808..5827
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5907..5982
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6044..6255
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6274..6283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6280..10383
                     /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             6280..10383
                     /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             10384..10431
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10432..10455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10465..10521
                     /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             10522..11118
                     /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     11015..11035
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11134..11722
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11187..11207)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11605..11616)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11794..12027
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12053..12070)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12059..12283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12329..12757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12416..12435)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12626..12647
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12766..12786)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(12771..13013,1..87)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(12951..13013,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     join(13013,1..19)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
       61 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
      121 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
      181 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
      241 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
      301 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
      361 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
      421 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
      481 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
      541 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
      601 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
      661 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
      721 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
      781 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
      841 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      901 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      961 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1021 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1081 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1141 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1201 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1261 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1321 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1381 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1441 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1501 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1561 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1621 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1681 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1741 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1801 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1861 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1921 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1981 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2041 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2101 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2161 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2221 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2281 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2341 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2401 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2461 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2521 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2581 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2641 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2701 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2761 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2821 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2881 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2941 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3001 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     3061 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     3121 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     3181 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     3241 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     3301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     3361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     3421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     3481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     3541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     3601 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     3661 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     3721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     3781 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     3841 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     3901 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     3961 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     4021 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     4081 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     4141 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     4201 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     4261 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     4321 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     4381 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     4441 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     4501 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     4561 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     4621 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     4681 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     4741 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     4801 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     4861 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     4921 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     4981 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     5041 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     5101 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     5161 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     5221 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     5281 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     5341 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     5401 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     5461 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     5521 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     5581 aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa ggtaccgagg
     5641 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     5701 ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     5761 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     5821 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     5881 acaccgggat gcctctcttg gccaaggttt tagagctaga aatagcaagt taaaataagg
     5941 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga attcgctagc
     6001 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     6061 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg
     6121 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     6181 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     6241 gccgccagaa cacaggaccg gttctagagc gctgccacca tggacaagaa gtacagcatc
     6301 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     6361 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     6421 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     6481 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     6541 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     6601 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     6661 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     6721 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     6781 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     6841 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     6901 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     6961 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     7021 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     7081 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     7141 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     7201 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     7261 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     7321 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     7381 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     7441 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     7501 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     7561 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     7621 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     7681 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     7741 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     7801 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     7861 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     7921 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     7981 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     8041 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     8101 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     8161 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     8221 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     8281 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     8341 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     8401 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     8461 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     8521 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     8581 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     8641 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     8701 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     8761 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     8821 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     8881 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     8941 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     9001 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     9061 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     9121 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     9181 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     9241 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     9301 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     9361 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     9421 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     9481 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     9541 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     9601 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     9661 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     9721 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     9781 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     9841 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     9901 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     9961 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
    10021 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
    10081 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
    10141 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
    10201 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
    10261 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
    10321 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
    10381 gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa agattacaaa
    10441 gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca agccggagat
    10501 gtcgaagaga atcctggacc gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
    10561 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
    10621 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
    10681 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
    10741 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
    10801 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
    10861 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc gcccgaccac
    10921 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
    10981 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
    11041 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
    11101 cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
    11161 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
    11221 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
    11281 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
    11341 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
    11401 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
    11461 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
    11521 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
    11581 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
    11641 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
    11701 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
    11761 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
    11821 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
    11881 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
    11941 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
    12001 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
    12061 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
    12121 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
    12181 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
    12241 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
    12301 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
    12361 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
    12421 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
    12481 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
    12541 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
    12601 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
    12661 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
    12721 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
    12781 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
    12841 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
    12901 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
    12961 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt ttt
//
