LOCUS       sequence-428772-       13577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for an inducible Cas9-P2A-Blasticidin 
            resistance casette with two sgRNA sequences against human TP73.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13577)
  TITLE     Selectable lentivectors delivering dual gRNAs and destabilized Cas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13577)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDual_dsCas9_Blast_hs_TP73
FEATURES             Location/Qualifiers
     source          1..13577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2608..2848
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     misc_RNA        2879..2954
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3045..3285
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     misc_RNA        3313..3388
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3450..3661
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3675..3684
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3681..3683,3684..4004)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             4014..8117
                     /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             4014..8117
                     /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             8118..8165
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8166..8189
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8199..8255
                     /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             8259..8654
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8669..9257
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8722..8742)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9140..9151)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9329..9562
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9588..9605)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9594..9818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9864..10292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9951..9970)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10161..10182
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10301..10321)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10306..10635
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10486..10621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10548..10567
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10683..10730
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10749..11123
                     /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    11253..11374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11290..11309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11344..11363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11423..11439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11423..11439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11436..11458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11447..11463
                     /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(11471..11477,11478..11495,11496..11501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11516..11537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11654..11671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11825..12413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11905..11924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12584..13375,13376..13444))
                     /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     13207..13226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13445..13549)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacctgcagc cttcggtttc cagccgcggt
     2881 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     2941 caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc
     3001 cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagagagggc ctatttccca
     3061 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     3121 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     3181 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     3241 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggaccgtct ctcagcatgg
     3301 acgtcttcca ccgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     3361 acttgaaaaa gtggcaccga gtcggtgctt ttttgaattc gctagctagg tcttgaaagg
     3421 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     3481 gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa
     3541 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     3601 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     3661 ggaccggttc tagagccacc atgggagtgc aggtggaaac catctcccca ggtgacgggc
     3721 gcaccttccc caagcgcggc cagacctgtg tggtgcacta caccgggatg cttgaagatg
     3781 gaaagaaagt cgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc
     3841 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
     3901 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
     3961 cacatgccac tctcgtcttc gatgtggagc ttctaaaacc ggaaccggct agcatggaca
     4021 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
     4081 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     4141 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     4201 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     4261 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     4321 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     4381 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     4441 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     4501 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     4561 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     4621 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     4681 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     4741 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     4801 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     4861 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     4921 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     4981 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     5041 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     5101 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     5161 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     5221 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     5281 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     5341 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     5401 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     5461 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     5521 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     5581 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     5641 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     5701 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     5761 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     5821 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     5881 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     5941 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     6001 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     6061 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     6121 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     6181 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     6241 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     6301 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     6361 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     6421 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     6481 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     6541 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     6601 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     6661 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     6721 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     6781 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     6841 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     6901 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     6961 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     7021 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     7081 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     7141 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     7201 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     7261 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     7321 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     7381 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     7441 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     7501 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     7561 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     7621 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     7681 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     7741 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     7801 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     7861 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     7921 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     7981 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     8041 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     8101 ctcagctggg aggcgacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga
     8161 agaaagatta caaagacgat gacgataagg gatccggagc tactaacttc agcctgctga
     8221 agcaggctgg ggacgtggag gagaaccctg gacctaggat ggccaagcct ttgtctcaag
     8281 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     8341 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     8401 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     8461 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     8521 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     8581 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     8641 atgtgtggga gggcgatcct aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     8701 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     8761 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     8821 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     8881 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     8941 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     9001 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     9061 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     9121 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     9181 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     9241 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     9301 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     9361 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9421 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9481 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9541 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     9601 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     9661 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     9721 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     9781 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     9841 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     9901 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     9961 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
    10021 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    10081 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    10141 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    10201 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    10261 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
    10321 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10381 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10441 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    10501 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    10561 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    10621 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    10681 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
    10741 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
    10801 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
    10861 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
    10921 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
    10981 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
    11041 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
    11101 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
    11161 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    11221 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    11281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    11341 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    11401 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    11461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    11521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    11581 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    11641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    11701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    11761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    11821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    11881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    11941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    12001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    12061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    12121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    12181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12241 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    12421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    12481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    12541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    12601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    12661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    12721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    12781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    12841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    12901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    12961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    13021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    13081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    13141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    13201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    13261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    13321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    13381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    13441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    13501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    13561 gaaaagtgcc acctgac
//
