LOCUS       Exported               12293 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of St1 dCas9-3XGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE-TO-nls-st1dCas9-3nls-3XGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12293)
  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 12293)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12293
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2986..3006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3016..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..7197
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             7216..7929
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             7948..8661
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    8683..9271
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8736..8756)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9154..9165)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9346..9579
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9698..9716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9784..9888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9889..10749
                     /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(10107..10126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10920..11508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11409..11428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11662..11679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     11771..11790
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    11919..11940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11955..11985
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11993..12009
                     /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     11998..12020
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12017..12033
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12017..12033
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12083..12102
                     /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 agtatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat
     6541 ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat
     6601 ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc ttggccgact
     6661 ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag
     6721 cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc
     6781 tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg
     6841 gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat
     6901 aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat
     6961 ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct
     7021 gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac
     7081 tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt
     7141 ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaatct
     7201 ggcggccgct ctggcatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg
     7261 gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt
     7321 gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta
     7381 ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg
     7441 gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa
     7501 cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa
     7561 ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat
     7621 atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat
     7681 aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc
     7741 gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg
     7801 ccagacaatc actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc
     7861 gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa
     7921 ctgtacaaag gtagcatcga tggtagcatg cgtaaaggcg aagagctgtt cactggtgtc
     7981 gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc
     8041 gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt
     8101 aaactgccgg taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt
     8161 gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc
     8221 tatgtgcagg aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa
     8281 gtgaaatttg aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa
     8341 gaagacggca atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac
     8401 atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg
     8461 gaggatggca gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt
     8521 cctgttctgc tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg
     8581 aacgagaaac gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat
     8641 ggtatggatg aactgtacaa actcgagtag ggtaatctag ataatcaacc tctggattac
     8701 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     8761 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     8821 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     8881 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     8941 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     9001 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     9061 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     9121 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     9181 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     9241 agtcggatct ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca
     9301 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     9361 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     9421 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     9481 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     9541 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     9601 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa
     9661 ttaaggaaag ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     9721 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     9781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     9841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     9901 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     9961 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
    10021 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
    10081 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
    10141 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
    10201 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
    10261 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
    10321 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
    10381 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
    10441 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
    10501 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
    10561 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
    10621 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
    10681 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
    10741 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
    10801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
    10861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
    10921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
    10981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
    11041 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
    11101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
    11161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
    11221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
    11281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
    11341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
    11401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
    11461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
    11521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
    11581 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
    11641 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
    11701 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac
    11761 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
    11821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg
    11881 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
    11941 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
    12001 ggataacaat ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca
    12061 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    12121 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
    12181 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
    12241 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
