LOCUS       sequence-309444-        9121 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 9121)
  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 9121)
  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-Tail-8A8G
FEATURES             Location/Qualifiers
     source          1..9121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..1265
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1256
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1213..1233
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1278..1874
                     /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(1278..1297)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1774..1794
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1875..1928
                     /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             1929..2624
                     /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    2646..3234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2699..2719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3117..3128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3306..3539
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3617..3738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3654..3673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3708..3727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3778..3913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3933..3952)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3934..3952)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3962..3979)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3962..3978)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3971..3993)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4120..4575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4207..4226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4417..4438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4601..4705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4706..4774,4775..5566)
                     /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(4924..4943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5737..6325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6226..6245
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6479..6496
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6613..6634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6649..6654,6655..6672,6673..6679)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6687..6703
                     /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     6692..6714
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6711..6727
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6711..6727
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6746..6766
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6748..6766
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6794..7020
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7021..7201
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7248..7373
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7866..8099
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8284..8328
                     /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        8506..8754
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8506..8526
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8677..8696
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    8952..9069
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
      421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg
     1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
     1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     1861 gcaagcccgg tgccgagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc
     1921 ccggccccgt gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca
     1981 ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     2041 cccagaccat gagaatcaag gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc
     2101 tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg
     2161 acttctttaa gcagtccttc cccgagggct tcacatggga gagagtcacc acatacgaag
     2221 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     2281 acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac
     2341 tcggctggga ggcctccacc gagaccctgt accccgctga cggcggcctg gaaggcagag
     2401 ccgacatggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttg aagaccacat
     2461 acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat gtggacagaa
     2521 gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac gaggtggctg
     2581 tggccagata ctgcgacctc cctagcaaac tggggcacag ataaggatcc acgcgttaag
     2641 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3181 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac
     3241 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     3301 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     3361 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     3421 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     3481 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     3541 tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     3601 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3661 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3721 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     3781 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3841 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3901 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     3961 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4021 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4081 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     4141 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4201 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4261 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4321 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4381 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4441 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4501 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4561 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4621 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4681 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4741 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4801 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4861 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4921 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4981 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5041 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5101 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5161 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5221 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5281 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5341 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5401 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5461 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5521 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5581 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5641 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5701 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5761 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5821 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5881 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5941 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6001 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6061 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6121 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6181 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6241 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6301 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6361 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6421 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6481 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6541 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6601 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6661 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6721 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     6781 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
     6841 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     6901 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
     6961 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
     7021 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7081 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7141 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7201 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     7261 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     7321 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     7381 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     7441 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     7501 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     7561 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     7621 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     7681 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     7741 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     7801 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     7861 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     7921 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     7981 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     8041 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     8101 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     8161 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     8221 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     8281 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     8341 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     8401 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     8461 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccagagaggg cctatttccc
     8521 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tagaattaat
     8581 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     8641 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     8701 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccggagac
     8761 gagtgatact cgtctctgtt tgagagctat gctggaaaca gcatagcaag ttcaaataag
     8821 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctcggaaa aaaaagaaaa
     8881 aaagaaaaaa agaaaaattt ttttgaattc cgtaactaga tcttgagaca aatggcagta
     8941 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     9001 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     9061 caaaattttc gggtttatta cagggacagc agagatccac tttggcgccg gctcgagggg
     9121 g
//
