LOCUS       sequence-332423-        9198 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 9198)
  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 9198)
  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-tRNA-Loop2-8A8G
FEATURES             Location/Qualifiers
     source          1..9198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..1274
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          327..1265
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1222..1242
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1287..1883
                     /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(1287..1306)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1783..1803
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1884..1937
                     /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             1938..2633
                     /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    2655..3243
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2708..2728)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3126..3137)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3315..3548
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3626..3747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3717..3736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3787..3922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3942..3961)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3943..3961)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3971..3988)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3971..3987)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3980..4002)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4129..4584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4216..4235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4426..4447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4610..4714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4715..4783,4784..5575)
                     /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(4933..4952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5746..6334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6235..6254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6488..6505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6622..6643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6658..6663,6664..6681,6682..6688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6696..6712
                     /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     6701..6723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6720..6736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6720..6736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6755..6775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6757..6775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6803..7029
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7030..7210
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7257..7382
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7875..8108
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8293..8337
                     /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        8515..8763
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8515..8535
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8686..8705
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    9038..9155
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tcgagggggc ccgggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct
       61 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg
      181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      301 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
      361 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
      421 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
      481 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
      541 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
      601 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
      661 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
      721 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
      781 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
      841 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
      901 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
      961 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1021 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1081 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1141 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1201 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1261 ttcaggtgtc gtgacgtacg gccaccatga ccgagtacaa gcccacggtg cgcctcgcca
     1321 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     1381 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     1441 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     1501 ccgtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     1561 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     1621 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc
     1681 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     1741 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     1801 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     1861 gcatgacccg caagcccggt gccgagggca ggggaagtct tctaacatgc ggggacgtgg
     1921 aggaaaatcc cggccccgtg agcgagctga ttaaggagaa catgcacatg aagctgtaca
     1981 tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct
     2041 acgagggcac ccagaccatg agaatcaagg cggtcgaggg cggccctctc cccttcgcct
     2101 tcgacatcct ggctaccagc ttcatgtacg gcagcaaaac cttcatcaac cacacccagg
     2161 gcatccccga cttctttaag cagtccttcc ccgagggctt cacatgggag agagtcacca
     2221 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     2281 tcatctacaa cgtcaagatc agaggggtga acttcccatc caacggccct gtgatgcaga
     2341 agaaaacact cggctgggag gcctccaccg agaccctgta ccccgctgac ggcggcctgg
     2401 aaggcagagc cgacatggcc ctgaagctcg tgggcggggg ccacctgatc tgcaacttga
     2461 agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc gtctactatg
     2521 tggacagaag actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg
     2581 aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaga taaggatcca
     2641 cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2701 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2761 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2821 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2881 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2941 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3001 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3061 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3121 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3181 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3241 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     3301 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     3361 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     3421 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     3481 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     3541 ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     3601 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     3661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3721 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     3781 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3841 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3901 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     3961 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4021 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4081 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     4141 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4201 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4261 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4321 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4381 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4441 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4501 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4561 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4621 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4681 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4741 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4801 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4861 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4921 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4981 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5041 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5101 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5161 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5221 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5281 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5341 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5401 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5461 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5521 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5581 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5641 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5701 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5761 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5821 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5881 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5941 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6001 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6061 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6121 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6181 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6241 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6301 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6361 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6421 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6481 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6541 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6601 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6661 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6721 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     6781 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
     6841 gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg
     6901 gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg
     6961 attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga
     7021 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     7081 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     7141 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     7201 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     7261 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     7321 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     7381 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     7441 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     7501 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     7561 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     7621 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     7681 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     7741 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     7801 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     7861 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     7921 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     7981 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     8041 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     8101 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     8161 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     8221 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     8281 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     8341 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     8401 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     8461 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cagagagggc
     8521 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
     8581 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
     8641 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
     8701 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
     8761 accgggttcc atggtgtaat ggttagcact ctggactctg aatccagcga tccgagttca
     8821 aatctcggtg gaacctagtg gcaggtgcac ctgcagtggt ttgagagcta tgctggaaac
     8881 agcatagcaa gttcaaataa ggctagtccg ttatcaactt ggccccgaaa aaaaagaaaa
     8941 aaagaaaaaa agaaaaaggg ccaagtggca ccgagtcggt gctttttttt gaattccgta
     9001 actagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     9061 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     9121 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     9181 atccactttg gcgccggc
//
