LOCUS       sequence-355759-        9194 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 9194)
  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 9194)
  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-Tail-8A8G
FEATURES             Location/Qualifiers
     source          1..9194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        99..1277
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          330..1268
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1225..1245
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1290..1886
                     /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(1290..1309)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1786..1806
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1887..1940
                     /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             1941..2636
                     /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    2658..3246
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2711..2731)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3129..3140)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3318..3551
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3629..3750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3666..3685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3720..3739
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3790..3925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3945..3964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3946..3964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3974..3991)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3974..3990)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3983..4005)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4132..4587
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4219..4238)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4429..4450
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4613..4717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4718..4786,4787..5578)
                     /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(4936..4955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5749..6337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6238..6257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6491..6508
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6625..6646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6661..6666,6667..6684,6685..6691)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6699..6715
                     /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     6704..6726
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6723..6739
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6723..6739
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6758..6778
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6760..6778
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6806..7032
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7033..7213
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7260..7385
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7878..8111
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8296..8340
                     /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        8518..8766
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8518..8538
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8689..8708
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    9037..9154
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctcgaggg ggcccgggtg caaagatgga taaagtttta aacagagagg aatctttgca
       61 gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
      121 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
      181 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
      241 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
      301 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
      361 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
      421 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
      481 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
      541 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
      601 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
      661 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
      721 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
      781 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
      841 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
      901 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
      961 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     1021 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     1081 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     1141 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     1201 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     1261 catttcaggt gtcgtgacgt acggccacca tgaccgagta caagcccacg gtgcgcctcg
     1321 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     1381 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     1441 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     1501 ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     1561 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     1621 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct
     1681 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     1741 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     1801 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     1861 ggtgcatgac ccgcaagccc ggtgccgagg gcaggggaag tcttctaaca tgcggggacg
     1921 tggaggaaaa tcccggcccc gtgagcgagc tgattaagga gaacatgcac atgaagctgt
     1981 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
     2041 cctacgaggg cacccagacc atgagaatca aggcggtcga gggcggccct ctccccttcg
     2101 ccttcgacat cctggctacc agcttcatgt acggcagcaa aaccttcatc aaccacaccc
     2161 agggcatccc cgacttcttt aagcagtcct tccccgaggg cttcacatgg gagagagtca
     2221 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     2281 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
     2341 agaagaaaac actcggctgg gaggcctcca ccgagaccct gtaccccgct gacggcggcc
     2401 tggaaggcag agccgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
     2461 tgaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     2521 atgtggacag aagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     2581 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac agataaggat
     2641 ccacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2701 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2761 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2821 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2881 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2941 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3001 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3061 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3121 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3181 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3241 ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta gccacttttt
     3301 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     3361 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     3421 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     3481 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     3541 aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc
     3601 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     3661 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3721 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     3781 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3841 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3901 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
     3961 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4021 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4081 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     4141 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4201 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4261 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4321 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4381 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4441 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4501 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4561 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     4621 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4681 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4741 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4801 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4861 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4921 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4981 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5041 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5101 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5161 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5221 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5281 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5341 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5401 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5461 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5521 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5581 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5641 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5701 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5761 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5821 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5881 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5941 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6001 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6061 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6121 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6181 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6241 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6301 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6361 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6421 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6481 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6541 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6601 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6661 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6721 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     6781 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
     6841 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
     6901 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
     6961 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
     7021 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7081 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7141 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7201 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     7261 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     7321 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     7381 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     7441 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     7501 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     7561 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     7621 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     7681 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     7741 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     7801 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     7861 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     7921 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     7981 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     8041 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     8101 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     8161 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     8221 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     8281 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     8341 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     8401 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     8461 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccagagag
     8521 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     8581 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     8641 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     8701 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     8761 aacaccgggt tccatggtgt aatggttagc actctggact ctgaatccag cgatccgagt
     8821 tcaaatctcg gtggaaccta gtggcaggtg cacctgcagt ggtttgagag ctatgctgga
     8881 aacagcatag caagttcaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     8941 tcggtgctcg gaaaaaaaag aaaaaaagaa aaaaagaaaa attttttttg aattccgtaa
     9001 ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     9061 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     9121 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     9181 tccactttgg cgcc
//
