LOCUS       sequence-71850-e        8998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian ST1-VP64 nuclease-null Cas9 activator expression, human 
            optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8998)
  AUTHORS   Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM
  TITLE     Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.
  JOURNAL   Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.
  PUBMED    24076762
REFERENCE   2  (bases 1 to 8998)
  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: "Nat 
            Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681."
COMMENT     Sequence Label: M-ST1n-VP64
FEATURES             Location/Qualifiers
     source          1..8998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(748..771,772..774,775..2538,2539..2544,2545..2610,
                     2611..2613,2614..4110)
                     /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="MSDLVLGLAIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLV
                     RRTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEE
                     LFIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYG
                     QLRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGK
                     RKYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLL
                     NDLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRID
                     KSGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADG
                     SFSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKT
                     TSSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNED
                     DEKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERC
                     LYTGKTISIHDLINNSNQFEVAAILPLSITFDDSLANKVLVYATAAQEKGQRTPYQALD
                     SMDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTRYASRVV
                     LNALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNL
                     WKKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSY
                     QVDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSK
                     FLMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGP
                     EIKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQ
                     FDKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRT
                     MPKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQ
                     HIIKNEGDKPKLDF"
     CDS             4123..4143
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4168..4317
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(4384..4403)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4428..4476
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4678..5106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4765..4784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4975..4996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5115..5135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5120..5449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5300..5435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5362..5381
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5516..6310
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5570..5589)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6180..6199
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6486..6607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6523..6542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6577..6596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6656..6672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6656..6672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6669..6691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6680..6696
                     /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(6704..6710,6711..6728,6729..6734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6749..6770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6887..6904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7058..7646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7138..7157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7817..8608,8609..8677))
                     /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     8440..8459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8678..8782)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8857..8876)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagg atcgaaccct tgccaccatg agcgacctgg tgctgggcct ggccatcggc
      781 atcggcagcg tgggcgtggg catcctgaac aaggtgaccg gcgagatcat ccacaagaac
      841 agtcgcatct tccctgctgc tcaggctgag aacaacctgg tgcgccgcac caaccgccag
      901 ggtcgccggc ttgctcgccg caagaagcac cggcgcgtgc gcctgaaccg cctgttcgag
      961 gagagcggcc tgatcaccga cttcaccaag atcagcatca acctgaaccc ctaccagctg
     1021 cgcgtgaagg gcctgaccga cgagctgagc aacgaggagc tgttcatcgc cctgaagaac
     1081 atggtgaagc accgcggcat cagctacctg gacgacgcca gcgacgacgg caacagcagc
     1141 gtgggcgact acgcccagat cgtgaaggag aacagcaagc agctggagac caagaccccc
     1201 ggccagatcc agctggagcg ctaccagacc tacggccagc tgcgcggcga cttcaccgtg
     1261 gagaaggacg gcaagaagca ccgcctgatc aacgtgttcc ccaccagcgc ctaccgcagc
     1321 gaggccctgc gcatcctgca gacccagcag gagttcaacc cccagatcac cgacgagttc
     1381 atcaaccgct acctggagat cctgaccggc aagcgcaagt actaccacgg ccccggcaac
     1441 gagaagagcc gcaccgacta cggccgctac cgcaccagcg gcgagaccct ggacaacatc
     1501 ttcggcatcc tgatcggcaa gtgcaccttc taccccgacg agttccgcgc cgccaaggcc
     1561 agctacaccg cccaggagtt caacctgctg aacgacctga acaacctgac cgtgcccacc
     1621 gagaccaaga agctgagcaa ggagcagaag aaccagatca tcaactacgt gaagaacgag
     1681 aaggccatgg gccccgccaa gctgttcaag tacatcgcca agctgctgag ctgcgacgtg
     1741 gccgacatca agggctaccg catcgacaag agcggcaagg ccgagatcca caccttcgag
     1801 gcctaccgca agatgaagac cctggagacc ctggacatcg agcagatgga ccgcgagacc
     1861 ctggacaagc tggcctacgt gctgaccctg aacaccgagc gcgagggcat ccaggaggcc
     1921 ctggagcacg agttcgccga cggcagcttc agccagaagc aggtggacga gctggtgcag
     1981 ttccgcaagg ccaacagcag catcttcggc aagggctggc acaacttcag cgtgaagctg
     2041 atgatggagc tgatccccga gctgtacgag accagcgagg agcagatgac catcctgacc
     2101 cgcctgggca agcagaagac caccagcagc agcaacaaga ccaagtacat cgacgagaag
     2161 ctgctgaccg aggagatcta caaccccgtg gtggccaaga gcgtgcgcca ggccatcaag
     2221 atcgtgaacg ccgccatcaa ggagtacggc gacttcgaca acatcgtgat cgagatggcc
     2281 cgcgagacca acgaggacga cgagaagaag gccatccaga agatccagaa ggccaacaag
     2341 gacgagaagg acgccgccat gctgaaggcc gccaaccagt acaacggcaa ggccgagctg
     2401 ccccacagcg tgttccacgg ccacaagcag ctggccacca agatccgcct gtggcaccag
     2461 cagggcgagc gctgcctgta caccggcaag accatcagca tccacgacct gatcaacaac
     2521 agcaaccagt tcgaggtggc tgccatcctg cccctgagca tcaccttcga cgacagcctg
     2581 gccaacaagg tgctggtgta cgccaccgcc gctcaggaga agggccagcg caccccctac
     2641 caggccctgg acagcatgga cgacgcctgg agcttccgcg agctgaaggc cttcgtgcgc
     2701 gagagcaaga ccctgagcaa caagaagaag gagtacctgc tgaccgagga ggacatcagc
     2761 aagttcgacg tgcgcaagaa gttcatcgag cgcaacctgg tggacacccg ctacgccagc
     2821 cgcgtggtgc tgaacgccct gcaggagcac ttccgcgccc acaagatcga caccaaggtg
     2881 agcgtggtgc gcggccagtt caccagccag ctgcgccgcc actggggcat cgagaagacc
     2941 cgcgacacct accaccacca cgccgtggac gccctgatca ttgcggcttc tagccagctg
     3001 aacctgtgga agaagcagaa gaacaccctg gtgagctaca gcgaggacca gctgctggac
     3061 atcgagaccg gcgagctgat cagcgacgac gagtacaagg agagcgtgtt caaggccccc
     3121 taccagcact tcgtggacac cctgaagagc aaggagttcg aggacagcat cctgttcagc
     3181 taccaggtgg acagcaagtt caaccgcaag atcagcgacg ccaccatcta cgccacccgc
     3241 caggccaagg tgggcaagga caaggccgac gagacctacg tgctgggcaa gatcaaggac
     3301 atctacaccc aggacggcta cgacgccttc atgaagatct acaagaagga caagagcaag
     3361 ttcctgatgt accgccacga cccccagacc ttcgagaagg tgatcgagcc catcctggag
     3421 aactacccca acaagcagat caacgataaa ggcaaggagg tgccctgcaa ccccttcctg
     3481 aagtacaagg aggagcacgg ctacatccgc aagtacagca agaagggcaa cggccccgag
     3541 atcaagagcc tgaagtacta cgacagcaag ctgggcaacc acatcgacat cacccccaag
     3601 gacagcaaca acaaggtggt gctgcagagc gtgagcccct ggcgcgccga cgtgtacttc
     3661 aacaagacca ccggcaagta cgagatcctg ggcctgaagt acgccgacct gcagtttgat
     3721 aagggcaccg gcacctacaa gatcagccag gagaagtaca acgacatcaa gaagaaggag
     3781 ggcgtggaca gcgacagcga gttcaagttc accctgtaca agaacgacct tctgctggtg
     3841 aaggacaccg agaccaagga gcaacagctg ttccgcttcc tgagccgcac catgcccaag
     3901 cagaagcact acgtggagct gaagccctac gacaagcaga agttcgaggg cggcgaggcc
     3961 ctgatcaagg tgctgggcaa cgtggccaac agcggccagt gcaagaaggg cctgggcaag
     4021 agcaacatca gcatctacaa ggtgcgcacc gacgtgctgg gcaaccagca catcatcaag
     4081 aacgagggcg acaagcccaa gctggacttc agcagggctg accccaagaa gaagaggaag
     4141 gtggaggcca gcggttccgg acgggctgac gcattggacg attttgatct ggatatgctg
     4201 ggaagtgacg ccctcgatga ttttgacctt gacatgcttg gttcggatgc ccttgatgac
     4261 tttgacctcg acatgctcgg cagtgacgcc cttgatgatt tcgacctgga catgctgatt
     4321 aactctagat gaaagggttc gatccctacc ggttagtaat gagtttaaac gggggaggct
     4381 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4441 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4501 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4561 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4621 gggcggcaga ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg
     4681 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4741 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4801 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     4861 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4921 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4981 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5041 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5101 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5221 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5281 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5341 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     5401 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     5461 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     5521 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5581 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5641 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     5701 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5761 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5821 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5881 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5941 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     6001 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     6061 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6121 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6181 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6241 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6301 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     6361 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6421 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6481 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6541 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6601 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6661 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6721 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6781 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6841 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     7261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7741 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7801 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7861 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7921 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7981 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8041 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8101 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8161 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8221 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8281 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8341 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8401 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8461 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8521 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8581 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8641 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8701 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8761 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8821 cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata
     8881 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8941 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
