LOCUS       sequence-108533-       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"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2299..2602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2603..2806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2757..2777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2808..2826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2857..2881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2974..2983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2986..3006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3016..3036,3037..3039,3040..4803,4804..4809,4810..4875,
                     4876..4878,4879..6375)
                     /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             6391..6411
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6424..6444
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6451..6471
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6484..7194
                     /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             7213..7923
                     /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             7942..8652
                     /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    8674..9262
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8727..8747)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9145..9156)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9337..9570
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9689..9707)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9775..9879
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9880..9948,9949..10740)
                     /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(10098..10117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10911..11499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11400..11419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11653..11670
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11910..11931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11946..11951,11952..11969,11970..11976)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11984..12000
                     /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     11989..12011
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12008..12024
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12008..12024
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12074..12093
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag
     2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga
     2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat
     2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     2941 cctaccggtt aaggtgaacg cgtgcaggct ggcgccacca tggcccctaa aaagaaacgc
     3001 aaggtcgaat tcggaagcga cctggtgctg ggcctggcca tcggcatcgg cagcgtgggc
     3061 gtgggcatcc tgaacaaggt gaccggcgag atcatccaca agaacagtcg catcttccct
     3121 gctgctcagg ctgagaacaa cctggtgcgc cgcaccaacc gccagggtcg ccggcttgct
     3181 cgccgcaaga agcaccggcg cgtgcgcctg aaccgcctgt tcgaggagag cggcctgatc
     3241 accgacttca ccaagatcag catcaacctg aacccctacc agctgcgcgt gaagggcctg
     3301 accgacgagc tgagcaacga ggagctgttc atcgccctga agaacatggt gaagcaccgc
     3361 ggcatcagct acctggacga cgccagcgac gacggcaaca gcagcgtggg cgactacgcc
     3421 cagatcgtga aggagaacag caagcagctg gagaccaaga cccccggcca gatccagctg
     3481 gagcgctacc agacctacgg ccagctgcgc ggcgacttca ccgtggagaa ggacggcaag
     3541 aagcaccgcc tgatcaacgt gttccccacc agcgcctacc gcagcgaggc cctgcgcatc
     3601 ctgcagaccc agcaggagtt caacccccag atcaccgacg agttcatcaa ccgctacctg
     3661 gagatcctga ccggcaagcg caagtactac cacggccccg gcaacgagaa gagccgcacc
     3721 gactacggcc gctaccgcac cagcggcgag accctggaca acatcttcgg catcctgatc
     3781 ggcaagtgca ccttctaccc cgacgagttc cgcgccgcca aggccagcta caccgcccag
     3841 gagttcaacc tgctgaacga cctgaacaac ctgaccgtgc ccaccgagac caagaagctg
     3901 agcaaggagc agaagaacca gatcatcaac tacgtgaaga acgagaaggc catgggcccc
     3961 gccaagctgt tcaagtacat cgccaagctg ctgagctgcg acgtggccga catcaagggc
     4021 taccgcatcg acaagagcgg caaggccgag atccacacct tcgaggccta ccgcaagatg
     4081 aagaccctgg agaccctgga catcgagcag atggaccgcg agaccctgga caagctggcc
     4141 tacgtgctga ccctgaacac cgagcgcgag ggcatccagg aggccctgga gcacgagttc
     4201 gccgacggca gcttcagcca gaagcaggtg gacgagctgg tgcagttccg caaggccaac
     4261 agcagcatct tcggcaaggg ctggcacaac ttcagcgtga agctgatgat ggagctgatc
     4321 cccgagctgt acgagaccag cgaggagcag atgaccatcc tgacccgcct gggcaagcag
     4381 aagaccacca gcagcagcaa caagaccaag tacatcgacg agaagctgct gaccgaggag
     4441 atctacaacc ccgtggtggc caagagcgtg cgccaggcca tcaagatcgt gaacgccgcc
     4501 atcaaggagt acggcgactt cgacaacatc gtgatcgaga tggcccgcga gaccaacgag
     4561 gacgacgaga agaaggccat ccagaagatc cagaaggcca acaaggacga gaaggacgcc
     4621 gccatgctga aggccgccaa ccagtacaac ggcaaggccg agctgcccca cagcgtgttc
     4681 cacggccaca agcagctggc caccaagatc cgcctgtggc accagcaggg cgagcgctgc
     4741 ctgtacaccg gcaagaccat cagcatccac gacctgatca acaacagcaa ccagttcgag
     4801 gtggctgcca tcctgcccct gagcatcacc ttcgacgaca gcctggccaa caaggtgctg
     4861 gtgtacgcca ccgccgctca ggagaagggc cagcgcaccc cctaccaggc cctggacagc
     4921 atggacgacg cctggagctt ccgcgagctg aaggccttcg tgcgcgagag caagaccctg
     4981 agcaacaaga agaaggagta cctgctgacc gaggaggaca tcagcaagtt cgacgtgcgc
     5041 aagaagttca tcgagcgcaa cctggtggac acccgctacg ccagccgcgt ggtgctgaac
     5101 gccctgcagg agcacttccg cgcccacaag atcgacacca aggtgagcgt ggtgcgcggc
     5161 cagttcacca gccagctgcg ccgccactgg ggcatcgaga agacccgcga cacctaccac
     5221 caccacgccg tggacgccct gatcattgcg gcttctagcc agctgaacct gtggaagaag
     5281 cagaagaaca ccctggtgag ctacagcgag gaccagctgc tggacatcga gaccggcgag
     5341 ctgatcagcg acgacgagta caaggagagc gtgttcaagg ccccctacca gcacttcgtg
     5401 gacaccctga agagcaagga gttcgaggac agcatcctgt tcagctacca ggtggacagc
     5461 aagttcaacc gcaagatcag cgacgccacc atctacgcca cccgccaggc caaggtgggc
     5521 aaggacaagg ccgacgagac ctacgtgctg ggcaagatca aggacatcta cacccaggac
     5581 ggctacgacg ccttcatgaa gatctacaag aaggacaaga gcaagttcct gatgtaccgc
     5641 cacgaccccc agaccttcga gaaggtgatc gagcccatcc tggagaacta ccccaacaag
     5701 cagatcaacg ataaaggcaa ggaggtgccc tgcaacccct tcctgaagta caaggaggag
     5761 cacggctaca tccgcaagta cagcaagaag ggcaacggcc ccgagatcaa gagcctgaag
     5821 tactacgaca gcaagctggg caaccacatc gacatcaccc ccaaggacag caacaacaag
     5881 gtggtgctgc agagcgtgag cccctggcgc gccgacgtgt acttcaacaa gaccaccggc
     5941 aagtacgaga tcctgggcct gaagtacgcc gacctgcagt ttgataaggg caccggcacc
     6001 tacaagatca gccaggagaa gtacaacgac atcaagaaga aggagggcgt ggacagcgac
     6061 agcgagttca agttcaccct gtacaagaac gaccttctgc tggtgaagga caccgagacc
     6121 aaggagcaac agctgttccg cttcctgagc cgcaccatgc ccaagcagaa gcactacgtg
     6181 gagctgaagc cctacgacaa gcagaagttc gagggcggcg aggccctgat caaggtgctg
     6241 ggcaacgtgg ccaacagcgg ccagtgcaag aagggcctgg gcaagagcaa catcagcatc
     6301 tacaaggtgc gcaccgacgt gctgggcaac cagcacatca tcaagaacga gggcgacaag
     6361 cccaagctgg acttcggctc tactagtggc cccaagaaga agaggaaggt aggaagtgga
     6421 agcccaaaga aaaagcggaa agtgggctca ccgaaaaaga agcgtaaggt tggatccgga
     6481 agtatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg
     6541 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
     6601 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     6661 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
     6721 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
     6781 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     6841 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     6901 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     6961 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag
     7021 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     7081 gactacagac tggaaagaat caaggaggcc aacaacgaaa cctacgtcga gcagcacgag
     7141 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattctggc
     7201 ggccgctctg gcatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag
     7261 ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc
     7321 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
     7381 ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac
     7441 acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga
     7501 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     7561 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg
     7621 atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc
     7681 ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca
     7741 aacgccaaga ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc
     7801 tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgaaac ctacgtcgag
     7861 cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt
     7921 aatggtagca tcgatggtag catggtgtct aagggcgaag agctgattaa ggagaacatg
     7981 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
     8041 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
     8101 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
     8161 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     8221 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     8281 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
     8341 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc
     8401 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
     8461 ctgatcgcaa acgccaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
     8521 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgaaacc
     8581 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
     8641 cacaagctta atctcgagta gggtaatcta gataatcaac ctctggatta caaaatttgt
     8701 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     8761 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     8821 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     8881 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     8941 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     9001 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     9061 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     9121 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     9181 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     9241 tccctttggg ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg
     9301 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     9361 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     9421 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     9481 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     9541 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     9601 atttataact tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa
     9661 gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     9721 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     9781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     9841 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     9901 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     9961 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
    10021 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
    10081 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
    10141 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
    10201 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
    10261 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
    10321 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
    10381 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
    10441 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
    10501 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
    10561 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
    10621 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
    10681 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
    10741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
    10801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
    10861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
    10921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
    10981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
    11041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
    11101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
    11161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
    11221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
    11281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
    11341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
    11401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
    11461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
    11521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
    11581 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
    11641 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
    11701 cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt
    11761 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
    11821 gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac
    11881 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    11941 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    12001 tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca
    12061 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
    12121 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
    12181 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
    12241 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt
//
