LOCUS       sequence-412471-       10526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing Cas9, GpNLuc, and sgRNA targeting 
            murine MLKL gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10526)
  AUTHORS   Cho MG, Kumar RJ, Lin CC, Boyer JA, Shahir JA, Fagan-Solis K, 
            Simpson DA, Fan C, Foster CE, Goddard AM, Lerner LM, Ellington SW, 
            Wang Q, Wang Y, Ho AY, Liu P, Perou CM, Zhang Q, McGinty RK, Purvis 
            JE, Gupta GP
  TITLE     MRE11 liberates cGAS from nucleosome sequestration during 
            tumorigenesis.
  JOURNAL   Nature. 2024 Jan 10. doi: 10.1038/s41586-023-06889-6.
  PUBMED    38200309
REFERENCE   2  (bases 1 to 10526)
  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: "Nature. 
            2024 Jan 10. doi: 10.1038/s41586-023-06889-6."
COMMENT     Sequence Label: Lenti-sgRNA-Cre-GpNLuc-sgmMLKL
FEATURES             Location/Qualifiers
     source          1..10526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     228..248
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(257..685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(367..388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     579..598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    731..955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     944..961
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             987..1220
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    1292..1880
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1398..1409
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1807..1827
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1896..2411)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             complement(join(2427..3137,3138..3140,3141..3143))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2427..3137,3138..3140,3141..3143))
                     /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(2469..2490)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2818..2837
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(3144..3200)
                     /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"
     CDS             complement(3201..4226)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             complement(4227..4247)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(4273..4484)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(4546..4621)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4642..4890)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4700..4719)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4870..4890)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    4941..5058
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5356..5400)
                     /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    5585..5818
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6311..6436
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6483..6663
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6681..6879)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6706..6726)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6881..7260
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7432..7451
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7526..7630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7631..7699,7700..8491)
                     /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     complement(7849..7868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8662..9250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9151..9170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9404..9421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9538..9559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9574..9579,9580..9597,9598..9604)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9612..9628
                     /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     9617..9639
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9636..9652
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9636..9652
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    9701..9822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9712..9731)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9766..9785
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(9952..10326)
                     /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"
     promoter        complement(10345..10392)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(10440..10526,1..243))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(10454..10526,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(10508..10526,1))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      241 cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
      301 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
      361 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
      421 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
      481 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
      541 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
      601 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
      661 cttaatgcgc cgctacaggg cgcgtgggga taccccctag agccccagct ggttctttcc
      721 gcctcagaag ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc
      781 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
      841 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
      901 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag
      961 gctgatcagc gggtttaaac gggccctgct agagattttc cacactgact aaaagggtct
     1021 gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt gaagcactca
     1081 aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga gctcccaggc
     1141 tcagatctgg tctaaccaga gagacccagt acaagcaaaa agcagatctt gtcttcgttg
     1201 ggagtgaatt agcccttcca gtcccccctt ttcttttaaa aagtggctaa gatctacagc
     1261 tgccttgtaa gtcattggtc ttaaagtcga cgcggggagg cggcccaaag ggagatccga
     1321 ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca gagccggcag caggccgcgg
     1381 gaaggaaggt ccgctggatt gagggccgaa gggacgtagc agaaggacgt cccgcgcaga
     1441 atccaggtgg caacacaggc gagcagccaa ggaaaggacg atgatttccc cgacaacacc
     1501 acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag gcaggcggcg
     1561 atgagttccg ccgtggcaat agggaggggg aaagcgaaag tcccggaaag gagctgacag
     1621 gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa acacagtgca caccacgcca
     1681 cgttgcctga caacgggcca caactcctca taaagagaca gcaaccagga tttatacaag
     1741 gaggagaaaa tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg
     1801 tatccacata gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt
     1861 ttgtaatcca gaggttgatt gtcgacttaa cgcgtttacg ccagaatgcg ttcgcacagc
     1921 cgccagccgg tcactccgtt gatggttact cggaacagca gggagccgtc ggggttgatc
     1981 aggcgctcgt cgataatttt gttgccgttc cacagggtcc ctgttacagt gatctttttg
     2041 ccgtcgaaca cggcgatgcc ttcatacggc cgtccgaaat agtcgatcat gttcggcgta
     2101 accccgtcga ttaccagtgt gccatagtgc aggatcacct taaagtgatg atcatccaca
     2161 gggtacacca ccttaaaaat tttttcgatc tggcccattt ggtcgccgct cagaccttca
     2221 tacgggatga tgacatggat gtcgatcttc agcccatttt caccgctcag gacaatcctt
     2281 tggatcggag ttacggacac cccgagattc tgaaacaaac tggacacacc tccctgttca
     2341 aggacttggt ccaggttgta gccggctgtc tgtcgccagt ccccaacgaa atcttcgagt
     2401 gtgaagacca tacctccgga gatatccttg tacagctcgt ccatgccgag agtgatcccg
     2461 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     2521 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     2581 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     2641 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     2701 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     2761 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     2821 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     2881 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     2941 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     3001 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     3061 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     3121 agctcctcgc ccttgctcac catcggtcca ggattctctt cgacatctcc ggcttgtttc
     3181 agcagagaga agtttgttgc atcgccatct tccagcaggc gcaccattgc ccctgtttca
     3241 ctatccaggt tacggatata gttcatgaca atatttacat tggtccagcc accagcttgc
     3301 atgatctccg gtattgaaac tccagcgcgg gccatatctc gcgcggctcc gacacgggca
     3361 ctgtgtccag accaggccag gtatctctga ccagagtcat ccttagcgcc gtaaatcaat
     3421 cgatgagttg cttcaaaaat cccttccagg gcgcgagttg atagctggct ggtggcagat
     3481 ggcgcggcaa caccattttt tctgacccgg caaaacaggt agttattcgg atcatcagct
     3541 acaccagata cggaaatcca tcgctcgacc agtttagtta cccccaggct aagtgccttc
     3601 tctacacctg cggtgctaac cagcgttttc gttctgccaa tatggattaa cattctccca
     3661 ccgtcagtac gtgagatatc tttaaccctg atcctggcaa tttcggctat acgtaacagg
     3721 gtgttataag caatccccag aaatgccaga ttacgtatat cctggcagcg atcgctattt
     3781 tccatgagtg aacgaacctg gtcgaaatca gtgcgttcga acgctagagc ctgttttgca
     3841 cgttcaccgg catcaacgtt ttcttttcgg atccgccgca taaccagtga aacagcattg
     3901 ctgtcacttg gtcgtggcag cccggaccga cgatgaagca tgtttagctg gcccaaatgt
     3961 tgctggatag tttttactgc cagaccgcgc gcctgaagat atagaagata atcgcgaaca
     4021 tcttcaggtt ctgcgggaaa ccatttccgg ttattcaact tgcaccatgc cgcccacgac
     4081 cggcaaacgg actgaagcat tttccaggta tgctcagaaa acgcctggcg atccctgaac
     4141 atgtccatca ggttcttgcg aacctcatca ctcgttgcat cgaccggtaa tgcaggcaaa
     4201 ttttggtgta cggtcagtaa attggacacc ttcctcttct tcttgggcat ggtagatcct
     4261 ctagaaccgg tcctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac
     4321 tactgcactt atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac
     4381 atcactttcc cagtttaccc cgcgccacct tctctaggca ccggatcaat tgccgacccc
     4441 tccccccaac ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga
     4501 gccaattccc actcctttca agacctagct agcgaattca aaaaagcacc gactcggtgc
     4561 cactttttca agttgataac ggactagcct tattttaact tgctatttct agctctaaaa
     4621 cgcgtctagg aaaccgtgtg cggtgtttcg tcctttccac aagatatata aagccaagaa
     4681 atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac ataattttaa
     4741 aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt
     4801 tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc gtatatgcaa
     4861 atatgaagga atcatgggaa ataggccctc ggtaccttaa ttaaccaaac tggatctctg
     4921 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
     4981 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
     5041 cccccttttc ttttaaaatt gtggatgaat actgccattt gtctgcagaa ttggcgcacg
     5101 cagtgccgat ccgttcacta atcgaatgga tctgtctctg tctctctctc caccttcttc
     5161 ttctattcct tcgggcctgt cgggtcccct cggggttggg aggtgggtct gaaacgataa
     5221 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     5281 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     5341 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     5401 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     5461 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     5521 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     5581 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     5641 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     5701 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     5761 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     5821 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     5881 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     5941 atctcctcct ccaggtctga agatcagcgg ccgcttgctg tgcggtggtc ttacttttgt
     6001 tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat
     6061 gcacacaata gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct
     6121 gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc
     6181 caggattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat
     6241 attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc
     6301 gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt
     6361 ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag
     6421 tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc
     6481 actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     6541 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     6601 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     6661 cccagtacag gcaaaacgcg ctgcttatat agacctccca ccgtacacgc ctaccgccca
     6721 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     6781 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     6841 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     6901 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     6961 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     7021 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     7081 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     7141 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     7201 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
     7261 aacgcgtata tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat
     7321 tcttcatgca attgtcggtc aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact
     7381 actcagcgac ctccaacaca caagcaggga gcagatactg gcttaactat gcggcatcag
     7441 agcagattgt actgagagtg caccataggg gatcgggaga tctcccgatc cgtcgacgtc
     7501 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     7561 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     7621 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7681 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7741 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7801 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7861 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7921 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7981 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     8041 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     8101 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     8161 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     8221 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     8281 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     8341 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     8401 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     8461 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     8521 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     8581 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     8641 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     8701 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     8761 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     8821 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8881 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8941 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     9001 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     9061 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     9121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     9181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     9241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     9301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     9361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     9421 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     9481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     9541 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     9601 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     9661 cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt
     9721 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     9781 attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc
     9841 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     9901 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtagcacgt gtcagtcctg
     9961 ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc
    10021 ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga
    10081 cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag
    10141 ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg
    10201 gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca
    10261 gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt
    10321 ggccatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
    10381 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
    10441 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
    10501 gaggcggcct cggcctctgc ataaat
//
