LOCUS       sequence-156208-       12284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of St1 dCas9-3XTagBFP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12284)
  AUTHORS   Ma H, Naseri A, Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T
  TITLE     Multicolor CRISPR labeling of chromosomal loci in human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7. doi: 
            10.1073/pnas.1420024112. Epub 2015 Feb 23.
  PUBMED    25713381
REFERENCE   2  (bases 1 to 12284)
  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"; doi: 
            "10.1073/pnas.1420024112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-10- 10"; volume: "112"; issue: "10"; pages: "3002-7"
COMMENT     Sequence Label: pHAGE-TO-nls-st1dCas9-3nls-3XTagBFP2
FEATURES             Location/Qualifiers
     source          1..12284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     255..279
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      372..381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             384..404
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(414..434,435..437,438..2201,2202..2207,2208..2273,
                     2274..2276,2277..3773)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus thermophilus Type II 
                     CRISPR/Cas system"
                     /label=St1dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D9A, D598A, 
                     H599A, and N622A"
                     /translation="SDLVLGLAIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLVR
                     RTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEEL
                     FIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYGQ
                     LRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGKR
                     KYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLLN
                     DLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRIDK
                     SGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADGS
                     FSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKTT
                     SSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNEDD
                     EKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERCL
                     YTGKTISIHDLINNSNQFEVAAILPLSITFDDSLANKVLVYATAAQEKGQRTPYQALDS
                     MDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTRYASRVVL
                     NALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNLW
                     KKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSYQ
                     VDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSKF
                     LMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGPE
                     IKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQF
                     DKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRTM
                     PKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQH
                     IIKNEGDKPKLDF"
     CDS             3789..3809
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3822..3842
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3849..3869
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3882..4592
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             4611..5321
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             5340..6050
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    6072..6660
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6125..6145)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6543..6554)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6735..6968
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7087..7105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7173..7277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7278..7346,7347..8138)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7496..7515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8309..8897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8798..8817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9051..9068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9308..9329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9344..9349,9350..9367,9368..9374)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9382..9398
                     /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)."
     primer_bind     9387..9409
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9406..9422
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9406..9422
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9472..9491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9683..10316
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10363..10488
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10985..11218
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11403..11447
                     /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    11740..11857
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        11981..12284
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
      241 agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      301 tgttttgacc tccatagaag acaccgggac cgatccagcc taccggttaa ggtgaacgcg
      361 tgcaggctgg cgccaccatg gcccctaaaa agaaacgcaa ggtcgaattc ggaagcgacc
      421 tggtgctggg cctggccatc ggcatcggca gcgtgggcgt gggcatcctg aacaaggtga
      481 ccggcgagat catccacaag aacagtcgca tcttccctgc tgctcaggct gagaacaacc
      541 tggtgcgccg caccaaccgc cagggtcgcc ggcttgctcg ccgcaagaag caccggcgcg
      601 tgcgcctgaa ccgcctgttc gaggagagcg gcctgatcac cgacttcacc aagatcagca
      661 tcaacctgaa cccctaccag ctgcgcgtga agggcctgac cgacgagctg agcaacgagg
      721 agctgttcat cgccctgaag aacatggtga agcaccgcgg catcagctac ctggacgacg
      781 ccagcgacga cggcaacagc agcgtgggcg actacgccca gatcgtgaag gagaacagca
      841 agcagctgga gaccaagacc cccggccaga tccagctgga gcgctaccag acctacggcc
      901 agctgcgcgg cgacttcacc gtggagaagg acggcaagaa gcaccgcctg atcaacgtgt
      961 tccccaccag cgcctaccgc agcgaggccc tgcgcatcct gcagacccag caggagttca
     1021 acccccagat caccgacgag ttcatcaacc gctacctgga gatcctgacc ggcaagcgca
     1081 agtactacca cggccccggc aacgagaaga gccgcaccga ctacggccgc taccgcacca
     1141 gcggcgagac cctggacaac atcttcggca tcctgatcgg caagtgcacc ttctaccccg
     1201 acgagttccg cgccgccaag gccagctaca ccgcccagga gttcaacctg ctgaacgacc
     1261 tgaacaacct gaccgtgccc accgagacca agaagctgag caaggagcag aagaaccaga
     1321 tcatcaacta cgtgaagaac gagaaggcca tgggccccgc caagctgttc aagtacatcg
     1381 ccaagctgct gagctgcgac gtggccgaca tcaagggcta ccgcatcgac aagagcggca
     1441 aggccgagat ccacaccttc gaggcctacc gcaagatgaa gaccctggag accctggaca
     1501 tcgagcagat ggaccgagag accctggaca agctggccta cgtgctgacc ctgaacaccg
     1561 agcgcgaggg catccaggag gccctggagc acgagttcgc cgacggcagc ttcagccaga
     1621 agcaggtgga cgagctggtg cagttccgca aggccaacag cagcatcttc ggcaagggct
     1681 ggcacaactt cagcgtgaag ctgatgatgg agctgatccc cgagctgtac gagaccagcg
     1741 aggagcagat gaccatcctg acccgcctgg gcaagcagaa gaccaccagc agcagcaaca
     1801 agaccaagta catcgacgag aagctgctga ccgaggagat ctacaacccc gtggtggcca
     1861 agagcgtgcg ccaggccatc aagatcgtga acgccgccat caaggagtac ggcgacttcg
     1921 acaacatcgt gatcgagatg gcccgcgaga ccaacgagga cgacgagaag aaggccatcc
     1981 agaagatcca gaaggccaac aaggacgaga aggacgccgc catgctgaag gccgccaacc
     2041 agtacaacgg caaggccgag ctgccccaca gcgtgttcca cggccacaag cagctggcca
     2101 ccaagatccg cctgtggcac cagcagggcg agcgctgcct gtacaccggc aagaccatca
     2161 gcatccacga cctgatcaac aacagcaacc agttcgaggt ggctgccatc ctgcccctga
     2221 gcatcacctt cgacgacagc ctggccaaca aggtgctggt gtacgccacc gccgctcagg
     2281 agaagggcca gcgcaccccc taccaggccc tggacagcat ggacgacgcc tggagcttcc
     2341 gcgagctgaa ggccttcgtg cgcgagagca agaccctgag caacaagaag aaggagtacc
     2401 tgctgaccga ggaggacatc agcaagttcg acgtgcgcaa gaagttcatc gagcgcaacc
     2461 tggtggacac ccgctacgcc agccgcgtgg tgctgaacgc cctgcaggag cacttccgcg
     2521 cccacaagat cgacaccaag gtgagcgtgg tgcgcggcca gttcaccagc cagctgcgcc
     2581 gccactgggg catcgagaag acccgcgaca cctaccacca ccacgccgtg gacgccctga
     2641 tcattgcggc ttctagccag ctgaacctgt ggaagaagca gaagaacacc ctggtgagct
     2701 acagcgagga ccagctgctg gacatcgaga ccggcgagct gatcagcgac gacgagtaca
     2761 aggagagcgt gttcaaggcc ccctaccagc acttcgtgga caccctgaag agcaaggagt
     2821 tcgaggacag catcctgttc agctaccagg tggacagcaa gttcaaccgc aagatcagcg
     2881 acgccaccat ctacgccacc cgccaggcca aggtgggcaa ggacaaggcc gacgagacct
     2941 acgtgctggg caagatcaag gacatctaca cccaggacgg ctacgacgcc ttcatgaaga
     3001 tctacaagaa ggacaagagc aagttcctga tgtaccgcca cgacccccag accttcgaga
     3061 aggtgatcga gcccatcctg gagaactacc ccaacaagca gatcaacgat aaaggcaagg
     3121 aggtgccctg caaccccttc ctgaagtaca aggaggagca cggctacatc cgcaagtaca
     3181 gcaagaaggg caacggcccc gagatcaaga gcctgaagta ctacgacagc aagctgggca
     3241 accacatcga catcaccccc aaggacagca acaacaaggt ggtgctgcag agcgtgagcc
     3301 cctggcgcgc cgacgtgtac ttcaacaaga ccaccggcaa gtacgagatc ctgggcctga
     3361 agtacgccga cctgcagttt gataagggca ccggcaccta caagatcagc caggagaagt
     3421 acaacgacat caagaagaag gagggcgtgg acagcgacag cgagttcaag ttcaccctgt
     3481 acaagaacga ccttctgctg gtgaaggaca ccgagaccaa ggagcaacag ctgttccgct
     3541 tcctgagccg caccatgccc aagcagaagc actacgtgga gctgaagccc tacgacaagc
     3601 agaagttcga gggcggcgag gccctgatca aggtgctggg caacgtggcc aacagcggcc
     3661 agtgcaagaa gggcctgggc aagagcaaca tcagcatcta caaggtgcgc accgacgtgc
     3721 tgggcaacca gcacatcatc aagaacgagg gcgacaagcc caagctggac ttcggctcta
     3781 ctagtggccc caagaagaag aggaaggtag gaagtggaag cccaaagaaa aagcggaaag
     3841 tgggctcacc gaaaaagaag cgtaaggttg gatccggaag tatggtgtct aagggcgaag
     3901 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc
     3961 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa
     4021 tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc
     4081 tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt
     4141 ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga
     4201 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag
     4261 gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct
     4321 tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga
     4381 agctcgtggg cgggagccat ctgatcgcaa acgccaagac cacatataga tccaagaaac
     4441 ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca
     4501 aggaggccaa caacgaaacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg
     4561 acctccctag caaactgggg cacaagctta atggtagcat cgatggtagc atggtgtcta
     4621 agggcgaaga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg
     4681 acaaccatca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga
     4741 ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta
     4801 ctagcttcct ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct
     4861 tcaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg
     4921 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca
     4981 agatcagagg ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct
     5041 gggaggcctt caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca
     5101 tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa cgccaagacc acatatagat
     5161 ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg
     5221 aaagaatcaa ggaggccaac aacgaaacct acgtcgagca gcacgaggtg gcagtggcca
     5281 gatactgcga cctccctagc aaactggggc acaagcttaa ttctggcggc cgctctggca
     5341 tggtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg
     5401 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     5461 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     5521 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
     5581 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     5641 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     5701 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
     5761 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
     5821 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac gccaagacca
     5881 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact
     5941 acagactgga aagaatcaag gaggccaaca acgaaaccta cgtcgagcag cacgaggtgg
     6001 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat ctcgagtaga
     6061 gtaatctaga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6121 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6181 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6241 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6301 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6361 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6421 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     6481 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6541 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6601 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6661 ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6721 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     6781 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6841 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6901 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6961 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     7021 aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat tcttgaagac
     7081 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     7141 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7201 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7261 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7321 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7381 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7441 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7501 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     7561 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7621 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     7681 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     7741 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7801 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7861 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7921 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7981 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     8041 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8101 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     8341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     8461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     8521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     8581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     8641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     8701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     8761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     8821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     8881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     8941 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     9001 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     9061 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     9121 ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc
     9181 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     9241 ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     9301 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     9361 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     9421 acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     9481 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     9541 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     9601 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
     9661 tatgtactac aaacttagta gttggaaggg ctaattcact cccaaagaag acaagatatc
     9721 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     9781 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     9841 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     9901 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     9961 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
    10021 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
    10081 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
    10141 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    10201 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    10261 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
    10321 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
    10381 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
    10441 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
    10501 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
    10561 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
    10621 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
    10681 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
    10741 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
    10801 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
    10861 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
    10921 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
    10981 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
    11041 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
    11101 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
    11161 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
    11221 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
    11281 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
    11341 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
    11401 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
    11461 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
    11521 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
    11581 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
    11641 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
    11701 ggtatcgccg aattgacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
    11761 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
    11821 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
    11881 agatccagtt tggactagcg atgctagctg atgcgggccc tagttattaa tagtaatcaa
    11941 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
    12001 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
    12061 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
    12121 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
    12181 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
    12241 ctacttggca gtacatctac gtattagtca tcgctattac catg
//
