LOCUS       sequence-413006-       14989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral Expression Plasmid for CRISPRoff2.1 driven by SFFV 
            Promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14989)
  AUTHORS   Allen TP, Roennfeldt AE, Reckdharajkumar M, Liu M, Quinn RJ, Russell
            DL, Peet DJ, Whitelaw ML, Bersten DC
  TITLE     dFLASH; dual FLuorescent transcription factor Activity Sensor for 
            Histone integrated live-cell reporting and high-content screening
  JOURNAL   bioRxiv 2023.11.21.568191
REFERENCE   2  (bases 1 to 14989)
  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: "bioRxiv 
            2023.11.21.568191"
COMMENT     Sequence Label: pLV-SFFVp-CRISPRoffv2.1-IRES-BlastR
FEATURES             Location/Qualifiers
     source          1..14989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    159..180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(195..200,201..218,219..225)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    233..249
                     /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     238..260
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     257..273
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     257..273
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     292..312
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        294..312
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        340..566
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             567..747
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    794..919
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1412..1645
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1830..1874
                     /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    2136..2253
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2281..2688
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     protein_bind    2754..2778
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(4737..4763,4764..4766,4767..7253,7254..7256,
                     7257..8840)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8844..8870
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8850..8876
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             8889..8909
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8916..8936
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8976..9671
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             9717..9902
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     protein_bind    complement(9932..9956)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    10101..10563
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(10157..10174)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     10384..10403
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             10567..10965
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             12198..12431
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    12503..12624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12540..12559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12594..12613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     12726..12745
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(13010..13028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13096..13200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13201..13269,13270..14061)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(13419..13438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      14232..14820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14721..14740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgataaccct aattcgatag catatgcttc ccgttgggta acatatgcta ttgaattagg
       61 gttagtctgg atagtatata ctactacccg ggaagcatat gctacccgtt tagggttcac
      121 cggtgatgcc ggccacgatg cgtccggcgt agaggatcta atgtgagtta gctcactcat
      181 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      241 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      301 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      361 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      421 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      481 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      541 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      601 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      661 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
      721 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
      781 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
      841 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
      901 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
      961 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1021 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1081 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1141 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1201 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1261 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1321 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1381 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1441 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1501 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1561 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1621 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1681 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     1741 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1801 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     1861 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     1921 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     1981 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2041 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2101 ccattcgatt agtgaacgga tctcgacggt atcggtttta aaagaaaagg ggggattggg
     2161 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     2221 ttacaaaaac aaattacaaa aattcaaaat tttatcgata ggtttggcaa gctagctgca
     2281 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca
     2341 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt
     2401 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc
     2461 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga
     2521 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc
     2581 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac
     2641 aacccctcac tcggcgcgcc agtcctccga cagactgagt cgcccggggg ggatctggag
     2701 ctctcgagaa ttctcacgcg actagtaacg gccgccagtg tgctggaatt atcacaagtt
     2761 tgtacaaaaa agcaggcttt aaaggaacca attcagtcga ctggatccgg taccgaattc
     2821 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatgaac catgaccagg
     2881 aatttgaccc cccaaaggtt tacccacctg tgccagctga gaagaggaag cccatccgcg
     2941 tgctgtctct ctttgatggg attgctacag ggctcctggt gctgaaggac ctgggcatcc
     3001 aagtggaccg ctacattgcc tccgaggtgt gtgaggactc catcacggtg ggcatggtgc
     3061 ggcaccaggg aaagatcatg tacgtcgggg acgtccgcag cgtcacacag aagcatatcc
     3121 aggagtgggg cccattcgac ctggtgattg gaggcagtcc ctgcaatgac ctctccattg
     3181 tcaaccctgc ccgcaaggga ctttatgagg gtactggccg cctcttcttt gagttctacc
     3241 gcctcctgca tgatgcgcgg cccaaggagg gagatgatcg ccccttcttc tggctctttg
     3301 agaatgtggt ggccatgggc gttagtgaca agagggacat ctcgcgattt cttgagtcta
     3361 accccgtgat gattgacgcc aaagaagtgt ctgctgcaca cagggcccgt tacttctggg
     3421 gtaaccttcc tggcatgaac aggcctttgg catccactgt gaatgataag ctggagctgc
     3481 aagagtgtct ggagcacggc agaatagcca agttcagcaa agtgaggacc attaccacca
     3541 ggtcaaactc tataaagcag ggcaaagacc agcatttccc cgtcttcatg aacgagaagg
     3601 aggacatcct gtggtgcact gaaatggaaa gggtgtttgg cttccccgtc cactacacag
     3661 acgtctccaa catgagccgc ttggcgaggc agagactgct gggccgatcg tggagcgtgc
     3721 cggtcatccg ccacctcttc gctccgctga aggaatattt tgcttgtgtg tctagcggca
     3781 atagtaacgc taacagccgc gggccgagct tcagcagcgg cctggtgccg ttaagcttgc
     3841 gcggcagcca tatgggccct atggagatat acaagacagt gtctgcatgg aagagacagc
     3901 cagtgcgggt actgagcctc ttcagaaaca tcgacaaggt actaaagagt ttgggcttct
     3961 tggaaagcgg ttctggttct gggggaggaa cgctgaagta cgtggaagat gtcacaaatg
     4021 tcgtgaggag agacgtggag aaatggggcc cctttgacct ggtgtacggc tcgacgcagc
     4081 ccctaggcag ctcttgtgat cgctgtcccg gctggtacat gttccagttc caccggatcc
     4141 tgcagtatgc gctgcctcgc caggagagtc agcggccctt cttctggata ttcatggaca
     4201 atctgctgct gactgaggat gaccaagaga caactacccg cttccttcag acagaggctg
     4261 tgaccctcca ggatgtccgt ggcagagact accagaatgc tatgcgggtg tggagcaaca
     4321 ttccagggct gaagagcaag catgcgcccc tgaccccaaa ggaagaagag tatctgcaag
     4381 cccaagtcag aagcaggagc aagctggacg ccccgaaagt tgacctcctg gtgaagaact
     4441 gccttctccc gctgagagag tacttcaagt atttttctca aaactcactt cctcttggag
     4501 ggccgagctc tggcgcaccc ccaccaagtg gagggtctcc tgccgggtcc ccaacatcta
     4561 ctgaagaagg caccagcgaa tccgcaacgc ccgagtcagg ccctggtacc tccacagaac
     4621 catctgaagg tagtgcgcct ggttccccag ctggaagccc tacttccacc gaagaaggca
     4681 cgtcaaccga accaagtgaa ggatctgccc ctgggaccag cactgaacca tctgagatgg
     4741 acaagaagta ttctatcgga ctggccatcg ggactaatag cgtcgggtgg gccgtgatca
     4801 ctgacgagta caaggtgccc tctaagaagt tcaaggtgct cgggaacacc gaccggcatt
     4861 ccatcaagaa aaatctgatc ggagctctcc tctttgattc aggggagacc gctgaagcaa
     4921 cccgcctcaa gcggactgct agacggcggt acaccaggag gaagaaccgg atttgttacc
     4981 ttcaagagat attctccaac gaaatggcaa aggtcgacga cagcttcttc cataggctgg
     5041 aagaatcatt cctcgtggaa gaggataaga agcatgaacg gcatcccatc ttcggtaata
     5101 tcgtcgacga ggtggcctat cacgagaaat acccaaccat ctaccatctt cgcaaaaagc
     5161 tggtggactc aaccgacaag gcagacctcc ggcttatcta cctggccctg gcccacatga
     5221 tcaagttcag aggccacttc ctgatcgagg gcgacctcaa tcctgacaat agcgatgtgg
     5281 ataaactgtt catccagctg gtgcagactt acaaccagct ctttgaagag aaccccatca
     5341 atgcaagcgg agtcgatgcc aaggccattc tgtcagcccg gctgtcaaag agccgcagac
     5401 ttgagaatct tatcgctcag ctgccgggtg aaaagaaaaa tggactgttc gggaacctga
     5461 ttgctctttc acttgggctg actcccaatt tcaagtctaa tttcgacctg gcagaggatg
     5521 ccaagctgca actgtccaag gacacctatg atgacgatct cgacaacctc ctggcccaga
     5581 tcggtgacca atacgccgac cttttccttg ctgctaagaa tctttctgac gccatcctgc
     5641 tgtctgacat tctccgcgtg aacactgaaa tcaccaaggc ccctctttca gcttcaatga
     5701 ttaagcggta tgatgagcac caccaggacc tgaccctgct taaggcactc gtccggcagc
     5761 agcttccgga gaagtacaag gaaatcttct ttgaccagtc aaagaatgga tacgccggct
     5821 acatcgacgg aggtgcctcc caagaggaat tttataagtt tatcaaacct atccttgaga
     5881 agatggacgg caccgaagag ctcctcgtga aactgaatcg ggaggatctg ctgcggaagc
     5941 agcgcacttt cgacaatggg agcattcccc accagatcca tcttggggag cttcacgcca
     6001 tccttcggcg ccaagaggac ttctacccct ttcttaagga caacagggag aagattgaga
     6061 aaattctcac tttccgcatc ccctactacg tgggacccct cgccagagga aatagccggt
     6121 ttgcttggat gaccagaaag tcagaagaaa ctatcactcc ctggaacttc gaagaggtgg
     6181 tggacaaggg agccagcgct cagtcattca tcgaacggat gactaacttc gataagaacc
     6241 tccccaatga gaaggtcctg ccgaaacatt ccctgctcta cgagtacttt accgtgtaca
     6301 acgagctgac caaggtgaaa tatgtcaccg aagggatgag gaagcccgca ttcctgtcag
     6361 gcgaacaaaa gaaggcaatt gtggaccttc tgttcaagac caatagaaag gtgaccgtga
     6421 agcagctgaa ggaggactat ttcaagaaaa ttgaatgctt cgactctgtg gagattagcg
     6481 gggtcgaaga tcggttcaac gcaagcctgg gtacctacca tgatctgctt aagatcatca
     6541 aggacaagga ttttctggac aatgaggaga acgaggacat ccttgaggac attgtcctga
     6601 ctctcactct gttcgaggac cgggaaatga tcgaggagag gcttaagacc tacgcccatc
     6661 tgttcgacga taaagtgatg aagcaactta aacggagaag atataccgga tggggacgcc
     6721 ttagccgcaa actcatcaac ggaatccggg acaaacagag cggaaagacc attcttgatt
     6781 tccttaagag cgacggattc gctaatcgca acttcatgca acttatccat gatgattccc
     6841 tgacctttaa ggaggacatc cagaaggccc aagtgtctgg acaaggtgac tcactgcacg
     6901 agcatatcgc aaatctggct ggttcacccg ctattaagaa gggtattctc cagaccgtga
     6961 aagtcgtgga cgagctggtc aaggtgatgg gtcgccataa accagagaac attgtcatcg
     7021 agatggccag ggaaaaccag actacccaga agggacagaa gaacagcagg gagcggatga
     7081 aaagaattga ggaagggatt aaggagctcg ggtcacagat ccttaaagag cacccggtgg
     7141 aaaacaccca gcttcagaat gagaagctct atctgtacta ccttcaaaat ggacgcgata
     7201 tgtatgtgga ccaagagctt gatatcaaca ggctctcaga ctacgacgtg gacgccatcg
     7261 tccctcagag cttcctcaaa gacgactcaa ttgacaataa ggtgctgact cgctcagaca
     7321 agaaccgggg aaagtcagat aacgtgccct cagaggaagt cgtgaaaaag atgaagaact
     7381 attggcgcca gcttctgaac gcaaagctga tcactcagcg gaagttcgac aatctcacta
     7441 aggctgagag gggcggactg agcgaactgg acaaagcagg attcattaaa cggcaacttg
     7501 tggagactcg gcagattact aaacatgtcg cccaaatcct tgactcacgc atgaatacca
     7561 agtacgacga aaacgacaaa cttatccgcg aggtgaaggt gattaccctg aagtccaagc
     7621 tggtcagcga tttcagaaag gactttcaat tctacaaagt gcgggagatc aataactatc
     7681 atcatgctca tgacgcatat ctgaatgccg tggtgggaac cgccctgatc aagaagtacc
     7741 caaagctgga aagcgagttc gtgtacggag actacaaggt ctacgacgtg cgcaagatga
     7801 ttgccaaatc tgagcaggag atcggaaagg ccaccgcaaa gtacttcttc tacagcaaca
     7861 tcatgaattt cttcaagacc gaaatcaccc ttgcaaacgg tgagatccgg aagaggccgc
     7921 tcatcgagac taatggggag actggcgaaa tcgtgtggga caagggcaga gatttcgcta
     7981 ccgtgcgcaa agtgctttct atgcctcaag tgaacatcgt gaagaaaacc gaggtgcaaa
     8041 ccggaggctt ttctaaggaa tcaatcctcc ccaagcgcaa ctccgacaag ctcattgcaa
     8101 ggaagaagga ttgggaccct aagaagtacg gcggattcga ttcaccaact gtggcttatt
     8161 ctgtcctggt cgtggctaag gtggaaaaag gaaagtctaa gaagctcaag agcgtgaagg
     8221 aactgctggg tatcaccatt atggagcgca gctccttcga gaagaaccca attgactttc
     8281 tcgaagccaa aggttacaag gaagtcaaga aggaccttat catcaagctc ccaaagtata
     8341 gcctgttcga actggagaat gggcggaagc ggatgctcgc ctccgctggc gaacttcaga
     8401 agggtaatga gctggctctc ccctccaagt acgtgaattt cctctacctt gcaagccatt
     8461 acgagaagct gaaggggagc cccgaggaca acgagcaaaa gcaactgttt gtggagcagc
     8521 ataagcatta tctggacgag atcattgagc agatttccga gttttctaaa cgcgtcattc
     8581 tcgctgatgc caacctcgat aaagtcctta gcgcatacaa taagcacaga gacaaaccaa
     8641 ttcgggagca ggctgagaat atcatccacc tgttcaccct caccaatctt ggtgcccctg
     8701 ccgcattcaa gtacttcgac accaccatcg accggaaacg ctatacctcc accaaagaag
     8761 tgctggacgc caccctcatc caccagagca tcaccggact ttacgaaact cggattgacc
     8821 tctcacagct cggaggggat gcctatccct atgacgtgcc cgattatgcc agcctgggca
     8881 gcggctcccc caagaaaaaa cgcaaggtgg aagatcctaa gaaaaagcgg aaagtggacg
     8941 gcattggtag tgggagcaac ggcagcagcg gatccagcga gctgattaag gagaacatgc
     9001 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg
     9061 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
     9121 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca
     9181 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat
     9241 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     9301 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg
     9361 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg
     9421 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc
     9481 tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc
     9541 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct
     9601 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc
     9661 acaagcttaa tggcggtggc ggagggatgg atgctaagtc actaactgcc tggtcccgga
     9721 cactggtgac cttcaaggat gtatttgtgg acttcaccag ggaggagtgg aagctgctgg
     9781 acactgctca gcagatcgtg tacagaaatg tgatgctgga gaactataag aacctggttt
     9841 ccttgggtta tcagcttact aagccagatg tgatcctccg gttggagaag ggagaagagc
     9901 cctagtaagc ggccgcactc gagatatcta gacccagctt tcttgtacaa agtggtgata
     9961 attctgcaga tatccatcac actggcggcc gctcgagcgc ccctctcccc taacgttact
    10021 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
    10081 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
    10141 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
    10201 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
    10261 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
    10321 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
    10381 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
    10441 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
    10501 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
    10561 taatccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
    10621 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
    10681 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
    10741 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
    10801 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
    10861 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
    10921 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagctag cctagtgcca
    10981 tttgttcagt ggttcgtagg gctttccccc actgtttggc tttcagttat atggatgatg
    11041 tggtattggg ggccaagtct gtacagcatc ttgagtccct ttttaccgct gttaccaatt
    11101 ttcttttgtc tttgggtata catttaaacc ctaacaaaac aaagagatgg ggttactctc
    11161 taaattttat gggttatgtc attggatgtt atgggtcctt gccacaagaa cacatcatac
    11221 aaaaaatcaa agaatgtttt agaaaacttc ctattaacag gcctattgat tggaaagtat
    11281 gtcaacgaat tgtgggtctt ttgggttttg ctgccccttt tacacaatgt ggttatcctg
    11341 cgttgatgcc tttgtatgca tgtattcaat ctaagcaggc tttcactttc tcgccaactt
    11401 acaaggcctt tctgtgtaaa caatacctga acctttaccc cgttgcccgg caacggccag
    11461 gtctgtgcca agtgtttgct gacgcaaccc ccactggctg gggcttggtc atgggccatc
    11521 agcgcatgcg tggaaccttt tcggctcctc tgccgatcca tactgcggaa ctcctagccg
    11581 cttgttttgc tcgcagcagg tctggagcaa acattatcgg gactgataac tctgttgtcc
    11641 tatcccgcaa atatacatcg tttccatggc tgctaggctg tgctgccaac tggatcctgc
    11701 gcgggacgtc ctttgtttac gtcccgtcgg cgctgaatcc tgcggacgac ccttctcggg
    11761 gtcgcttggg actctctcgt ccccttctcc gtctgccgtt ccgaccgacc acggggcgca
    11821 cctctcttta cgcggactcc ccgtctgtgc cttctcatct gccggaccgt gtgcacttcg
    11881 cttcacctct gcacgtcgca tggagaccac cgtgaacgcc caccaaatat tgcccaaggt
    11941 cttacataag aggactcttg gactctcagc aatgtcaacg accgaccttg aggcatactt
    12001 caaagactgt ttgtttaaag actgggagga gttgggggag gagattaggt taaaggtctt
    12061 tgtactagga ggctgtaggc ataaattggt ctgcgcacca gcaccatggc gcaatcacta
    12121 gagcggggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
    12181 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
    12241 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    12301 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    12361 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    12421 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
    12481 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
    12541 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    12601 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
    12661 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
    12721 ttttttattt atgcagaggc cgaggccgga tcccttgagt ggctttcatc ctggagcaga
    12781 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
    12841 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
    12901 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
    12961 tagcaatagt gtttataagg cccccttgtt aattcttgaa gacgaaaggg cctcgtgata
    13021 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
    13081 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
    13141 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
    13201 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
    13261 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
    13321 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
    13381 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
    13441 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
    13501 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
    13561 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
    13621 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
    13681 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
    13741 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
    13801 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
    13861 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
    13921 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
    13981 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
    14041 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
    14101 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
    14161 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
    14221 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
    14281 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
    14341 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
    14401 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
    14461 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
    14521 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
    14581 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
    14641 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
    14701 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
    14761 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
    14821 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttg aagctgtccc
    14881 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
    14941 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcg
//
