LOCUS       sequence-275145-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human TRRAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Kwan SY, Sheel A, Song CQ, Zhang XO, Jiang T, Dang H, Cao Y, Ozata 
            DM, Mou H, Yin H, Weng Z, Wang XW, Xue W
  TITLE     Depletion of TRRAP induces p53-independent senescence in liver 
            cancer by downregulating mitotic genes.
  JOURNAL   Hepatology. 2019 Jun 12. doi: 10.1002/hep.30807.
  PUBMED    31188495
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: "Hepatology.
            2019 Jun 12. doi: 10.1002/hep.30807."
COMMENT     Sequence Label: TRRAP sgRNA1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             18..198
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    245..370
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    863..1096
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1281..1325
                     /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    1623..1740
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1791..2039
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1791..1811
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1962..1981
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2061..2136
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2198..2409
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2428..2437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2434..6537
                     /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             2434..6537
                     /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             6538..6585
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6586..6609
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6619..6675
                     /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             6676..7272
                     /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     7169..7189
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7288..7876
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7341..7361)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7759..7770)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7948..8181
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8207..8224)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8213..8437
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8483..8911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8570..8589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8780..8801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8920..8940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8925..9254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9105..9240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9167..9186
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9302..9349
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9368..9742
                     /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    9872..9993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9909..9928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9963..9982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10042..10058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10042..10058)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10055..10077)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10066..10082
                     /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(10090..10096,10097..10114,10115..10120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10135..10156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10273..10290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10444..11032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10524..10543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11203..11994,11995..12063))
                     /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     11826..11845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12064..12168)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12243..12262)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12434..12813
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12815..13013
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     12968..12988
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
       61 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      121 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      181 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      241 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
      301 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
      361 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
      421 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
      481 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
      541 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
      601 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
      661 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
      721 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
      781 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
      841 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
      901 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
      961 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     1021 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     1081 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     1141 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     1201 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     1261 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     1321 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     1381 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     1441 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     1501 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     1561 tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac
     1621 aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1681 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1741 cgggtttatt acagggacag cagagatcca gtttggttaa ttaaggtacc gagggcctat
     1801 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa
     1861 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     1921 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     1981 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     2041 actcctgata caagagatcg gttttagagc tagaaatagc aagttaaaat aaggctagtc
     2101 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc
     2161 ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     2221 gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc
     2281 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     2341 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     2401 agaacacagg accggttcta gagcgctgcc accatggaca agaagtacag catcggcctg
     2461 gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     2521 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     2581 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     2641 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     2701 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     2761 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     2821 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     2881 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     2941 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     3001 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     3061 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     3121 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     3181 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     3241 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     3301 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     3361 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     3421 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     3481 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     3541 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     3601 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     3661 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     3721 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     3781 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     3841 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     3901 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     3961 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     4021 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     4081 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     4141 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     4201 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     4261 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     4321 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     4381 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     4441 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     4501 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     4561 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     4621 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     4681 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     4741 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     4801 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     4861 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     4921 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     4981 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     5041 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     5101 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     5161 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     5221 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     5281 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     5341 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     5401 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     5461 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     5521 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     5581 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     5641 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     5701 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     5761 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     5821 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     5881 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     5941 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     6001 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     6061 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     6121 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     6181 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     6241 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     6301 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     6361 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     6421 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     6481 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaag
     6541 cgacctgccg ccacaaagaa ggctggacag gctaagaaga agaaagatta caaagacgat
     6601 gacgataagg gatccggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa
     6661 gagaatcctg gaccgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     6721 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     6781 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     6841 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     6901 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     6961 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     7021 cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga ccaccagggc
     7081 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     7141 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     7201 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     7261 cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     7321 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     7381 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     7441 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     7501 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     7561 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     7621 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     7681 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     7741 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     7801 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     7861 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
     7921 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     7981 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     8041 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     8101 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     8161 cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct
     8221 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     8281 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     8341 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     8401 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     8461 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     8521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     8581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     8641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     8701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     8761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     8821 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     8881 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     8941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     9001 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     9061 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     9121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     9181 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     9241 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     9301 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     9361 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     9421 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     9481 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     9541 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     9601 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     9661 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     9721 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     9781 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     9841 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     9901 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     9961 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
    10021 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    10081 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    10141 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    10201 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    10261 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    10321 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    10381 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    10441 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    10501 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    10561 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    10621 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    10681 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    10741 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    10801 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    10861 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
    10921 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    10981 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    11041 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    11101 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
    11161 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
    11221 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
    11281 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
    11341 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    11401 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    11461 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    11521 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    11581 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    11641 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    11701 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    11761 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    11821 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    11881 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    11941 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    12001 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    12061 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    12121 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    12181 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
    12241 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
    12301 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
    12361 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
    12421 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
    12481 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
    12541 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
    12601 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
    12661 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
    12721 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
    12781 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
    12841 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
    12901 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
    12961 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cag
//
