LOCUS       sequence-332449-        9190 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 9190)
  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 9190)
  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-Tetra Loop-8A8G
FEATURES             Location/Qualifiers
     source          1..9190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    92..209
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        348..1526
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          579..1517
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1474..1494
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1539..2135
                     /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(1539..1558)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2035..2055
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2136..2189
                     /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             2190..2885
                     /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    2907..3495
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2960..2980)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3378..3389)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3567..3800
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3878..3999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3915..3934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3969..3988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4039..4174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4194..4213)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4195..4213)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4223..4240)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4223..4239)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4232..4254)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4381..4836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4468..4487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4678..4699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4862..4966
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4967..5035,5036..5827)
                     /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(5185..5204)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5998..6586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6487..6506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6740..6757
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6874..6895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6910..6915,6916..6933,6934..6940)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6948..6964
                     /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     6953..6975
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6972..6988
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6972..6988
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7007..7027
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7009..7027
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7055..7281
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7282..7462
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7509..7634
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8127..8360
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8545..8589
                     /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        8767..9015
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8767..8787
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8938..8957
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttttgaattc cgtaactaga
       61 tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
      121 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
      181 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
      241 tttggcgccg gctcgagggg gcccgggtgc aaagatggat aaagttttaa acagagagga
      301 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
      361 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      421 attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      481 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      541 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
      601 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
      661 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      721 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
      781 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
      841 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
      901 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
      961 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     1021 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     1081 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     1141 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     1201 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     1261 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1321 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1381 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1441 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1501 tttttcttcc atttcaggtg tcgtgacgta cggccaccat gaccgagtac aagcccacgg
     1561 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     1621 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     1681 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     1741 acgacggcgc cgccgtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     1801 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     1861 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     1921 tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     1981 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     2041 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     2101 cgcgcacctg gtgcatgacc cgcaagcccg gtgccgaggg caggggaagt cttctaacat
     2161 gcggggacgt ggaggaaaat cccggccccg tgagcgagct gattaaggag aacatgcaca
     2221 tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg
     2281 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggcggtcgag ggcggccctc
     2341 tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaaa accttcatca
     2401 accacaccca gggcatcccc gacttcttta agcagtcctt ccccgagggc ttcacatggg
     2461 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     2521 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc
     2581 ctgtgatgca gaagaaaaca ctcggctggg aggcctccac cgagaccctg taccccgctg
     2641 acggcggcct ggaaggcaga gccgacatgg ccctgaagct cgtgggcggg ggccacctga
     2701 tctgcaactt gaagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg
     2761 gcgtctacta tgtggacaga agactggaaa gaatcaagga ggccgacaaa gagacctacg
     2821 tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca
     2881 gataaggatc cacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     2941 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3001 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3061 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3121 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3181 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3241 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3301 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3361 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3421 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3481 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
     3541 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     3601 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     3661 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     3721 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     3781 agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt
     3841 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     3901 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3961 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     4021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4141 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     4201 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4261 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4321 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     4381 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4441 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4501 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4561 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4621 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4681 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4741 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4801 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4921 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4981 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5041 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5101 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5161 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5221 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5281 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5341 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5401 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5461 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5521 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5581 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5641 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5701 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5761 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5821 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5881 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5941 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6001 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6061 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6121 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6241 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6301 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6361 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6421 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6481 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6541 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6601 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6661 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6721 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6781 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6841 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6901 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6961 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     7021 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     7081 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     7141 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     7201 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     7261 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     7321 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7381 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7441 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     7501 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     7561 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     7621 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     7681 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     7741 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     7801 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     7861 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     7921 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     7981 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     8041 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     8101 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     8161 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     8221 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     8281 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     8341 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     8401 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     8461 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     8521 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     8581 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     8641 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     8701 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     8761 cccagagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     8821 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     8881 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     8941 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     9001 aaaggacgaa acaccgggtt ccatggtgta atggttagca ctctggactc tgaatccagc
     9061 gatccgagtt caaatctcgg tggaacctag tggcaggtgc acctgcagtg gtttgagagc
     9121 tatgctggcg aaaaaaaaga aaaaaagaaa aaaagaaaaa ccagcatagc aagttcaaat
     9181 aaggctagtc
//
