LOCUS       sequence-413573-        8945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral backbone for expressing U6 driven hybrid guide (hg)RNAs 
            with BveI cloning sites, mCherry and puromycin selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8945)
  AUTHORS   Xiao MS, Damodaran AP, Kumari B, Dickson E, Xing K, On TA, Parab N, 
            King HE, Perez AR, Guiblet WM, Duncan G, Che A, Chari R, Andresson 
            T, Vidigal JA, Weatheritt RJ, Aregger M, Gonatopoulos-Pournatzis T
  TITLE     Genome-scale exon perturbation screens uncover exons critical for 
            cell fitness.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2553-2572.e19. doi: 
            10.1016/j.molcel.2024.05.024. Epub 2024 Jun 24.
  PUBMED    38917794
REFERENCE   2  (bases 1 to 8945)
  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"; doi: 
            "10.1016/j.molcel.2024.05.024"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2553-2572.e19"
COMMENT     Sequence Label: pLCHKOv3-mCherry
FEATURES             Location/Qualifiers
     source          1..8945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..507
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     426..444
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             529..1125
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(529..548)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1025..1045
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1141..1194
                     /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             1201..1908
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1343..1361)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1611..1630
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1923..2511
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1976..1996)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2394..2405)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2583..2816
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2894..3015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2985..3004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3055..3190
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3210..3229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3211..3229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3239..3256)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3239..3255)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3248..3270)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3397..3852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3484..3503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3694..3715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3878..3982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3983..4051,4052..4843)
                     /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(4201..4220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5014..5602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5503..5522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5756..5773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5890..5911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5926..5931,5932..5949,5950..5956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5964..5980
                     /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     5969..5991
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5988..6004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5988..6004
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6023..6043
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6025..6043
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6071..6297
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6298..6478
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6525..6650
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7143..7376
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7561..7605
                     /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"
     primer_bind     8333..8352
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        complement(8466..8714)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(8524..8543)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(8694..8714)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    8756..8873
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcagctc acactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat gaccgagtac
      541 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
      601 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
      661 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
      721 tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag cgtcgaagcg
      781 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
      841 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
      901 ctggccaccg tcggtgtttc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
      961 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     1021 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     1081 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccacgcg tggcagtgga
     1141 gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccactcgag
     1201 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1261 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1321 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1381 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     1441 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1501 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     1561 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1621 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     1681 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     1741 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     1801 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     1861 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaacg cgttaagtcg
     1921 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     1981 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2041 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2101 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2161 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2221 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2281 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2341 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2401 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2461 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt
     2521 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     2581 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     2641 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     2701 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     2761 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtac
     2821 gtatagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     2881 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2941 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3001 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
     3061 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3121 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3181 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     3241 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3301 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3361 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3421 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3481 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3541 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3601 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3661 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3721 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3781 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3841 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4201 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4501 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5161 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5581 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5641 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5701 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5761 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5821 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5881 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5941 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     6001 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     6061 gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt
     6121 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
     6181 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
     6241 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
     6301 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6361 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6421 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6481 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     6541 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     6601 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     6661 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     6721 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     6781 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     6841 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     6901 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     6961 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     7021 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     7081 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     7141 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     7201 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     7261 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     7321 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     7381 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     7441 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     7501 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     7561 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     7621 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     7681 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     7741 ttatcgtttc agacccacct cccaaccccg aggggaccct gcaggattta aatccgaggg
     7801 gacccagaga attcaaaaaa gtaggcaggt gcttactaaa agccagataa cagtatgcgt
     7861 atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     7921 caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     7981 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     8041 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     8101 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat
     8161 gcagtttaag ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     8221 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     8281 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     8341 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     8401 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaacctgc
     8461 agagcggtac ctcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa
     8521 gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca
     8581 agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa
     8641 ttaattctaa ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg
     8701 ggaaataggc cctcgaattc tagatcttga gacaaatggc agtattcatc cacaatttta
     8761 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     8821 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     8881 attacaggga cagcagagat ccactttggc gccggctcga gatttaaatc ctgcaggcga
     8941 ggggg
//
