LOCUS       sequence-384994-        9134 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Cas9-2A-Puro expression vector bearing a sgRNA targeting Exon 1 of 
            human lncRNA T-REX17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9134)
  AUTHORS   Landshammer A, Bolondi A, Kretzmer H, Much C, Buschow R, Rose A, Wu 
            HJ, Mackowiak SD, Braendl B, Giesselmann P, Tornisiello R, Parsi KM,
            Huey J, Mielke T, Meierhofer D, Maehr R, Hnisz D, Michor F, Rinn JL,
            Meissner A
  TITLE     T-REX17 is a transiently expressed non-coding RNA essential for 
            human endoderm formation.
  JOURNAL   Elife. 2023 Jan 31;12:e83077. doi: 10.7554/eLife.83077.
  PUBMED    36719724
REFERENCE   2  (bases 1 to 9134)
  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: "Elife."; 
            date: "2023-01-31"; volume: "12:e83077. doi"; pages: " 
            10.7554/eLife.83077"
COMMENT     Sequence Label: pX459_V2.0_pSpCas9(BB)-2A-Puro_T-REX17_Ex1_KI
FEATURES             Location/Qualifiers
     source          1..9134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          135..362
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      374..383
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             383..448
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             455..475
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             500..4600
                     /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             500..4600
                     /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             4601..4648
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4664..4717
                     /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             4718..5317
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4718..4737)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5345..5362)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5351..5558
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5567..5707
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5567..5696
                     /label=AAV2 ITR
     rep_origin      5782..6237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5869..5888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6079..6100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6254..6273)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6373..6395
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6433..6451)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6519..6623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6624..6692,6693..7484)
                     /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(6842..6861)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7655..8243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8144..8163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8305..8553
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8305..8325
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8476..8495
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8575..8650
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8747..9032
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggcggggg gsggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
       61 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      121 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
      181 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      241 acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt
      301 tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc
      361 aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga
      421 tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
      481 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
      541 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
      601 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
      661 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
      721 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
      781 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
      841 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
      901 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
      961 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     1021 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     1081 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     1141 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     1201 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     1261 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     1321 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     1381 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     1441 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     1501 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     1561 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     1621 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     1681 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     1741 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     1801 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     1861 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     1921 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     1981 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     2041 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     2101 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     2161 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     2221 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     2281 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     2341 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     2401 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     2461 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     2521 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     2581 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     2641 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     2701 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     2761 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     2821 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     2881 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     2941 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     3001 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     3061 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     3121 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     3181 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     3241 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     3301 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     3361 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     3421 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     3481 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     3541 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     3601 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     3661 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     3721 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     3781 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     3841 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     3901 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     3961 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     4021 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     4081 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     4141 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     4201 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     4261 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     4321 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     4381 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     4441 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     4501 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     4561 gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa
     4621 aaaggccggc caggcaaaaa agaaaaagga attcggcagt ggagagggca gaggaagtct
     4681 gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg accgagtaca agcccacggt
     4741 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4801 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4861 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4921 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4981 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5041 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5101 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5161 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccacaacct
     5221 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5281 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagaa ttctaactag agctcgctga
     5341 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     5401 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     5461 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     5521 ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg
     5581 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     5641 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     5701 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5761 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5821 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     5881 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5941 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     6001 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6061 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     6121 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     6181 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     6241 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     6301 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     6361 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     6421 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     6481 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6541 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6601 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6661 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6721 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6781 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6841 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6901 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6961 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7021 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7081 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7141 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7201 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7261 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7321 ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc
     7381 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7441 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7501 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7561 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7621 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7681 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7741 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7801 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7861 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7921 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7981 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8041 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8101 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8161 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8221 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8281 tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat
     8341 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
     8401 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     8461 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     8521 gctttatata tcttgtggaa aggacgaaac accgtgagag gaaccatctt cagggtttta
     8581 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     8641 gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     8701 tttagcgcgt gcgccaattc tgcagacaaa tggctctaga ggtacccgtt acataactta
     8761 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa
     8821 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     8881 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     8941 aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt
     9001 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     9061 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     9121 tttgtgcagc gatg
//
