LOCUS       sequence-332394-        9118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses programmed gRNA scaffold from U6 promoter. Programmed gRNA
            scaffold for streamlined scRNA-seq in CRISPR screen (Direct-seq). 
            3rd generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9118)
  AUTHORS   Song Q, Ni K, Liu M, Li Y, Wang L, Wang Y, Liu Y, Yu Z, Qi Y, Lu Z, 
            Ma L
  TITLE     Direct-seq: programmed gRNA scaffold for streamlined scRNA-seq in 
            CRISPR screen.
  JOURNAL   Genome Biol. 2020 Jun 8;21(1):136. doi: 10.1186/s13059-020-02044-w.
  PUBMED    32513233
REFERENCE   2  (bases 1 to 9118)
  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: "Genome 
            Biol."; date: "2020-06-8"; volume: "21(1):136. doi"; pages: " 
            10.1186/s13059-020-02044-w"
COMMENT     Sequence Label: Direct-seq_lentiGuide-Puro-Tetra Loop-8A8G
FEATURES             Location/Qualifiers
     source          1..9118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        115..1293
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          346..1284
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1241..1261
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1306..1902
                     /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(1306..1325)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1802..1822
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1903..1956
                     /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             1957..2652
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     misc_feature    2674..3262
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2727..2747)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3145..3156)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3334..3567
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3645..3766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3682..3701)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3736..3755
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3806..3941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3961..3980)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3962..3980)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3990..4007)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3990..4006)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3999..4021)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4148..4603
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4235..4254)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4445..4466
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4629..4733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4734..4802,4803..5594)
                     /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(4952..4971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5765..6353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6254..6273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6507..6524
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6641..6662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6677..6682,6683..6700,6701..6707)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6715..6731
                     /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     6720..6742
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6739..6755
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6739..6755
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6774..6794
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6776..6794
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6822..7048
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7049..7229
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7276..7401
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7894..8127
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8312..8356
                     /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"
     promoter        8534..8782
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8534..8554
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8705..8724
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    8977..9094
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gatccacttt ggcgccggct cgagggggcc cgggtgcaaa gatggataaa gttttaaaca
       61 gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc
      121 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      181 gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      241 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      301 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      361 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      421 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      481 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
      541 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      601 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
      661 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
      721 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
      781 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
      841 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
      901 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
      961 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1021 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1081 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1141 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1201 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1261 caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgac cgagtacaag
     1321 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     1381 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     1441 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     1501 gtcgcggacg acggcgccgc cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     1561 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     1621 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     1681 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     1741 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     1801 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     1861 gaaggaccgc gcacctggtg catgacccgc aagcccggtg ccgagggcag gggaagtctt
     1921 ctaacatgcg gggacgtgga ggaaaatccc ggccccgtga gcgagctgat taaggagaac
     1981 atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc
     2041 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggc ggtcgagggc
     2101 ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcaaaacc
     2161 ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc cgagggcttc
     2221 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
     2281 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc
     2341 aacggccctg tgatgcagaa gaaaacactc ggctgggagg cctccaccga gaccctgtac
     2401 cccgctgacg gcggcctgga aggcagagcc gacatggccc tgaagctcgt gggcgggggc
     2461 cacctgatct gcaacttgaa gaccacatac agatccaaga aacccgctaa gaacctcaag
     2521 atgcccggcg tctactatgt ggacagaaga ctggaaagaa tcaaggaggc cgacaaagag
     2581 acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg
     2641 gggcacagat aaggatccac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     2701 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2761 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2821 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2881 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2941 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3001 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3061 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3121 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3181 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3241 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     3301 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     3361 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     3421 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     3481 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     3541 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
     3601 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     3661 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3721 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     3781 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3841 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3901 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     3961 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     4021 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4081 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4141 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4201 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4261 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4321 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4381 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4441 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4501 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4561 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     4621 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4681 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4741 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4801 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4861 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4921 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4981 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5041 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5101 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5161 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5221 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5281 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5341 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5401 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5461 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5521 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5581 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5641 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5701 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5761 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5821 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5881 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5941 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6001 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6061 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6121 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6181 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6241 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6301 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6361 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6421 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6481 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6541 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6601 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     6661 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     6721 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     6781 accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact
     6841 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
     6901 gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
     6961 acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt
     7021 atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg
     7081 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7141 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7201 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     7261 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     7321 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     7381 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     7441 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     7501 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     7561 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     7621 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     7681 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     7741 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     7801 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     7861 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     7921 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     7981 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     8041 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     8101 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     8161 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     8221 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     8281 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     8341 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     8401 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     8461 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     8521 gaggggaccc agagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
     8581 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
     8641 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
     8701 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
     8761 cttgtggaaa ggacgaaaca ccggagacga gtgatactcg tctctgtttg agagctatgc
     8821 tggcgaaaaa aaagaaaaaa agaaaaaaag aaaaaccagc atagcaagtt caaataaggc
     8881 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttttg aattccgtaa
     8941 ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     9001 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     9061 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcaga
//
