LOCUS       sequence-449964-       15016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral Expression of dCas9-KRAB from S. iniae fused to the 
            repressive KRAB domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15016)
  AUTHORS   Butterfield GL, Rohm D, Roberts A, Nethery MA, Rizzo AJ, Morone DJ, 
            Garnier L, Iglesias N, Barrangou R, Gersbach CA
  TITLE     Characterization of diverse Cas9 orthologs for genome and epigenome 
            editing.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Mar 18;122(11):e2417674122. doi: 
            10.1073/pnas.2417674122. Epub 2025 Mar 12.
  PUBMED    40073054
REFERENCE   2  (bases 1 to 15016)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-03-18"; volume: "122(11):e2417674122. 
            doi"; pages: " 10.1073/pnas.2417674122. Epub 2025 Mar 12"
COMMENT     Sequence Label: pFUGW hUbC-Sin-dCas9-KRAB-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..15016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        225..423
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     378..398
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             441..621
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    668..793
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1286..1519
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1704..1748
                     /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    2046..2163
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2205..2238)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2262..3473
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3310..3329
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3502..3511
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3511..3576
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3586..3606
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7732..7752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7789..7974
                     /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"
     CDS             7981..8034
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(8041..8043,8044..8046,8047..8757)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8041..8043,8044..8046,8047..8757)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8086..8107)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     8694..8715
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8767..8823
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(9256..9289)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9314..9902
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9367..9387)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9785..9796)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9905..9921)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9906..9922)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10427..10607
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10633..10650)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10639..10863
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10909..11337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10996..11015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11206..11227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11346..11366)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11351..11680
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11531..11666
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11593..11612
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11728..11775
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11794..12168
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12298..12419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12335..12354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12389..12408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12468..12484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12468..12484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12481..12503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12492..12508
                     /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(12516..12522,12523..12540,12541..12546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12561..12582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12699..12716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12870..13458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12950..12969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13629..14420,14421..14489))
                     /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     14252..14271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14490..14594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14669..14688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(14860..15016,1..223)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
       61 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      121 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      181 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      241 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      301 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      361 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      421 cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
      481 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      541 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      601 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
      661 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
      721 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
      781 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
      841 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
      901 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
      961 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1021 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1081 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1141 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1201 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1261 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1321 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1381 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1441 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1501 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1561 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1621 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     1681 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     1741 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     1801 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     1861 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     1921 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     1981 gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc
     2041 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     2101 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     2161 tttcgggttt attacaggga cagcagagat ccagtttggt taatataact tcgtatagca
     2221 tacattatac gaagttatta acccgtgtcg gctccagatc tggcctccgc gccgggtttt
     2281 ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg
     2341 cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2401 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2461 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2521 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2581 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     2641 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     2701 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     2761 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     2821 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     2881 ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc
     2941 ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3001 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3061 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3121 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     3181 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3241 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3301 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     3361 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     3421 agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt
     3481 gggctgcagg tcgactctag agccaccatg gactacaaag accatgacgg tgattataaa
     3541 gatcatgaca tcgattacaa ggatgacgat gacaagcacg ttaaccccaa gaagaagagg
     3601 aaggtgggcc gcggaaccgg tatgcgcaaa ccttactcaa ttggcctggc aatcgggact
     3661 aattctgttg gctgggctgt gattactgat gattacaagg tgccaagtaa gaaaatgagg
     3721 attcagggca cgactgatcg gaccagcatt aagaagaatc tcattggggc cctcctgttc
     3781 gataatggcg agactgccga ggccactcga ttaaagagaa caacaaggag gaggtacacc
     3841 agacggaagt accgaataaa ggaactgcaa aagatcttca gcagcgaaat gaatgagctc
     3901 gacattgctt ttttccctag actgtctgag agttttcttg tgagtgacga caaagaattc
     3961 gagaatcatc cgatttttgg aaaccttaaa gatgagataa cttatcataa cgattaccct
     4021 actatttatc acttgcgaca gacacttgca gaccgtgacc agaaggccga tcttaggctc
     4081 atttatctcg ctctggccca cattattaaa tttcgggggc actttttgat cgaaggcaat
     4141 ctggacagtg agaacacgga cgtacacgtg ctgtttctga acctggtgaa catatataat
     4201 aacctgttcg aggaagatat agttgaaacc gcatccatag acgctgagaa gattcttacc
     4261 tcaaaaactt ccaaatccag gcggctcgag aatcttatag ctgagattcc taaccagaag
     4321 cggaacatgt tgtttggcaa cctcgtgtct ctggctctcg gcctgacacc aaattttaaa
     4381 accaattttg agctgctgga ggatgcaaag ttacagatct ccaaggattc atatgaagaa
     4441 gacctcgaca acttgttggc acagattggg gatcagtacg cagatctctt tatcgccgct
     4501 aaaaagcttt ctgacgcaat attactgtct gacatcatca ccgtgaaggg cgcctccact
     4561 aaagcgcctc tttcagcatc catggtgcag agatatgaag agcatcaaca ggacctcgct
     4621 ctcctgaaga atctcgtgaa aaaacagatt cctgagaagt ataaggaaat cttcgataac
     4681 aaggagaaga atggctatgc aggttatatc gatggcaaga cctcccagga ggaattttac
     4741 aagtacatca agcccatact tcttaagctc aacggcacag agaagttgat cagcaaactt
     4801 gagcgggagg acttcctgag aaagcaacga acattcgaca acggatctat tcctcaccag
     4861 attcacctga atgagctcaa ggcaatcatc cggaggcagg agaagtttta tccctttctg
     4921 aaggaaaatc agaagaaaat cgaaaagctt ttcacattta aaattcccta ttacgtcggg
     4981 ccactcgcca atggccagag tagcttcgcc tggctgaaga gacagtccaa cgagtctatc
     5041 accccctgga acttcgagga agtggtggat caagaggcct cagcgcgcgc cttcatagag
     5101 aggatgacta acttcgatac ctatttaccc gaggagaagg ttctgccaaa gcacagccca
     5161 ctctacgaaa tgtttatggt ctataatgag ctcaccaagg ttaagtatca gaccgagggg
     5221 atgaagaggc ccgtctttct ctcttccgaa gacaaagaag aaatagtgaa tctcctgttc
     5281 aaaaaagacc ggaaggtcac tgtcaagcag ctgaaggagg aatatttctc caaaatgaaa
     5341 tgcttccaca ccgtgacaat cttgggcgtg gaggatcggt ttaatgcttc tctgggcacg
     5401 taccatgacc tgctcaaaat ttttaaagat aaagccttct tagacgatga ggccaatcaa
     5461 gatatcttgg aagagatcgt atggacttta acgctttttg aggatcaagc catgattgaa
     5521 agaaggctgg tgaagtacgc ggacgtgttc gaaaaatccg tccttaaaaa gttaaagaaa
     5581 cgccattaca cgggctgggg acgtctttcc cagaagctta ttaatgggat caaagacaaa
     5641 caaactggga agacaattct cggctttctg aaagacgacg gtgtagccaa ccgaaatttt
     5701 atgcagttaa ttaacgacag ctccctggac ttcgcaaaga ttatcaagca tgaacaggaa
     5761 aaaaccatca agaacgagtc attggaggaa acgattgcga acctggcagg cagccccgcc
     5821 attaagaaag gcattcttca gtctattaaa attgtcgatg aaatcgttaa gattatggga
     5881 cagaacccag acaatattgt tattgagatg gcacgcgaga accaatccac gatgcaagga
     5941 atcaaaaact cccgacagcg tctgcgcaag ctcgaggagg tgcataagaa caccgggtcc
     6001 aagattttga aagaatacaa cgtgagtaat acgcagcttc agagcgatag gctctattta
     6061 tacctgctgc aggacggaaa ggatatgtac accggcaagg agttggacta cgacaatctt
     6121 agtcaatatg atattgatgc gatcatccct cagtctttca taaaagataa ctctatcgac
     6181 aacatagtgc tgactacaca agctagtaat aggggcaagt cagacaacgt gcccaacata
     6241 gagattgtga acaaaatgaa gtctttttgg tataaacagc tcaaaaatgg ggcaattagc
     6301 cagcgcaaat tcgaccattt aaccaaggcc gagcgtggcg cactgagcga tttcgataag
     6361 gcaggcttta tcaagcgcca gctcgtcgag acacggcaga taaccaaaca tgtggctcaa
     6421 atcctggaca gtcggttcaa ttccaatctt acggaggact ctaaatctaa cagaaacgtt
     6481 aagataataa ctctcaagtc aaaaatggtg agtgacttcc gaaaggactt tggcttttac
     6541 aagctgagag aagtaaatga ttatcaccac gcccaggacg catatctcaa tgccgtcgtc
     6601 ggtactgcct tacttaagaa gtaccctaaa ctggaagcag agttcgtgta tggggattac
     6661 aagcactacg atctcgctaa gttaatgatt caaccggaca gtagccttgg aaaagccaca
     6721 accagaatgt tcttctattc taacctcatg aatttcttca aaaaagaaat caaactggcc
     6781 gatgatacta tatttacgag gccccagatt gaagtgaaca ccgaaactgg ggagattgtc
     6841 tgggataagg taaaggacat gcagaccatc aggaaagtga tgtcctatcc acaagtcaac
     6901 atagtgatga aaaccgaagt ccagactggg gggttttcta aggagagtat cctgcctaag
     6961 ggaaactcag acaaactgat cgcccgcaag aaatcctggg accctaagaa atacggtggt
     7021 ttcgatagcc ctatcattgc atattcagtc ctggtcgtcg ctaagatagc caaaggcaaa
     7081 acccagaaac tcaagactat taaagagttg gtcggtatca aaatcatgga gcaggacgaa
     7141 ttcgaaaagg atccaattgc gtttctcgaa aagaagggct atcaggacat acagacctct
     7201 tccatcatca agctgccgaa gtactctctc tttgagcttg agaatggacg caagagactg
     7261 ctggctagcg ccaaagaact gcagaagggc aacgaactgg ccctccctaa caaatacgta
     7321 aagttcttgt atttagcatc tcattacaca aaattcacag gtaaggagga agatcgagaa
     7381 aaaaagcgct cctatgtaga gtcacacctg tattactttg acgagattat gcagattatc
     7441 gttgagtatt ctaaccggta cattctcgcc gacagcaatc tgattaaaat tcagaacttg
     7501 tacaaagaga aggataactt tagtatcgag gagcaagcca ttaatatgct caatctcttc
     7561 acttttacag atctcggcgc gccagccgct ttcaagttct ttaacggaga tatagatcgg
     7621 aagcggtaca gctctaccaa cgagatcatt aattctactc tgatttacca gagtcccaca
     7681 gggttatacg agaccaggat cgacctcagt aagctggggg gcaaaaccgg tcctaagaaa
     7741 aagcggaaag tggctagcat ggatgctaag tcactaactg cctggtcccg gacactggtg
     7801 accttcaagg atgtatttgt ggacttcacc agggaggagt ggaagctgct ggacactgct
     7861 cagcagatcg tgtacagaaa tgtgatgctg gagaactata agaacctggt ttccttgggt
     7921 tatcagctta ctaagccaga tgtgatcctc cggttggaga agggagaaga gcccgctagc
     7981 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctggacca
     8041 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     8101 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     8161 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     8221 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     8281 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     8341 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     8401 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     8461 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     8521 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     8581 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     8641 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     8701 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggga
     8761 tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga
     8821 ccgaagacct tcaacatctc tcagcaggat ctggagctgg tggaggtcgc cactgagaag
     8881 atcaccatgc tctatgagga caacaagcac catgtcgggg cggccatcag gaccaagact
     8941 ggggagatca tctctgctgt ccacattgag gcctacattg gcagggtcac tgtctgtgct
     9001 gaagccattg ccattgggtc tgctgtgagc aacgggcaga aggactttga caccattgtg
     9061 gctgtcaggc acccctactc tgatgaggtg gacagatcca tcagggtggt cagcccctgt
     9121 ggcatgtgca gagagctcat ctctgactat gctcctgact gctttgtgct cattgagatg
     9181 aatggcaagc tggtcaaaac caccattgag gaactcatcc ccctcaagta caccaggaac
     9241 taaagttgat aatagataac ttcgtatagc atacattata cgaagttatg aattcgatat
     9301 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     9361 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     9421 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     9481 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     9541 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     9601 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     9661 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     9721 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     9781 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     9841 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     9901 gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     9961 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
    10021 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
    10081 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
    10141 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
    10201 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
    10261 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
    10321 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
    10381 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
    10441 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
    10501 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
    10561 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa
    10621 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
    10681 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
    10741 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
    10801 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
    10861 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
    10921 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
    10981 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
    11041 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
    11101 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
    11161 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
    11221 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
    11281 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
    11341 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
    11401 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
    11461 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
    11521 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
    11581 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
    11641 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
    11701 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
    11761 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
    11821 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
    11881 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
    11941 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
    12001 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
    12061 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
    12121 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
    12181 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
    12241 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
    12301 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
    12361 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
    12421 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
    12481 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
    12541 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
    12601 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
    12661 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
    12721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
    12781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
    12841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
    12901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
    12961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
    13021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
    13081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
    13141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
    13201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
    13261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
    13321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
    13381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
    13441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
    13501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
    13561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
    13621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
    13681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
    13741 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
    13801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
    13861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
    13921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
    13981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
    14041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    14101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    14161 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    14221 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    14281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    14341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    14401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    14461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    14521 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    14581 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    14641 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    14701 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    14761 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
    14821 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
    14881 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
    14941 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
    15001 caataatgac gtatgt
//
