LOCUS       sequence-428810-       13881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for an inducible Cas9-P2A-BFP with two 
            sgRNA sequences against human non-coding DNA region of chromosome 1 
            (negative control).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13881)
  TITLE     Selectable lentivectors delivering dual gRNAs and destabilized Cas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13881)
  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            "Unpublished"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_BFP_hs_NC_chr1
FEATURES             Location/Qualifiers
     source          1..13881
                     /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             8256..8957
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    8973..9561
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9026..9046)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9444..9455)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9633..9866
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9892..9909)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9898..10122
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10168..10596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10255..10274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10465..10486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10605..10625)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10610..10939
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10790..10925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10852..10871
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10987..11034
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11053..11427
                     /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    11557..11678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11594..11613)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11648..11667
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11727..11743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11727..11743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11740..11762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11751..11767
                     /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(11775..11781,11782..11799,11800..11805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11820..11841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11958..11975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12129..12717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12209..12228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12888..13679,13680..13748))
                     /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     13511..13530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13749..13853)
                     /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 aacctgcatt gttgttatag tgtgagtggt
     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 ctagaaatca
     3301 tcctagactt aagttttaga 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 gatccggtgc cacgaatttc tcactactca
     8221 agcaggccgg tgatgtcgag gaaaaccctg gtcctatgag cgagctgatt aaggagaaca
     8281 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
     8341 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     8401 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
     8461 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
     8521 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     8581 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
     8641 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgaa actctgtacc
     8701 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
     8761 atctgatcgc aaacgccaag accacatata gatccaagaa acccgctaag aacctcaaga
     8821 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga
     8881 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
     8941 ggcacaagct taattgagga tcctaagtcg acaatcaacc tctggattac aaaatttgtg
     9001 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     9061 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     9121 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     9181 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     9241 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     9301 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     9361 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     9421 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     9481 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     9541 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
     9601 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     9661 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     9721 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     9781 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     9841 ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg
     9901 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     9961 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
    10021 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
    10081 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
    10141 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
    10201 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
    10261 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
    10321 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
    10381 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
    10441 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
    10501 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
    10561 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
    10621 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
    10681 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
    10741 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
    10801 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
    10861 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
    10921 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
    10981 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
    11041 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
    11101 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
    11161 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
    11221 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
    11281 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
    11341 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
    11401 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
    11461 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
    11521 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
    11581 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
    11641 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
    11701 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
    11761 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
    11821 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
    11881 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
    11941 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
    12001 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
    12061 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
    12121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
    12181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
    12241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
    12301 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
    12361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
    12421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
    12481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
    12541 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
    12601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
    12661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
    12721 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
    12781 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
    12841 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
    12901 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
    12961 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
    13021 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
    13081 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
    13141 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
    13201 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
    13261 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
    13321 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
    13381 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
    13441 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
    13501 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
    13561 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
    13621 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
    13681 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
    13741 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
    13801 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
    13861 ccccgaaaag tgccacctga c
//
