LOCUS       sequence-203144-       10053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA expression in mammalian cells after Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10053)
  AUTHORS   Zhu S, Li W, Liu J, Chen CH, Liao Q, Xu P, Xu H, Xiao T, Cao Z, Peng
            J, Yuan P, Brown M, Liu XS, Wei W
  TITLE     Genome-scale deletion screening of human long non-coding RNAs using 
            a paired-guide RNA CRISPR-Cas9 library.
  JOURNAL   Nat Biotechnol. 2016 Dec;34(12):1279-1286. doi: 10.1038/nbt.3715. 
            Epub 2016 Oct 31.
  PUBMED    27798563
REFERENCE   2  (bases 1 to 10053)
  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"; doi: "10.1038/nbt.3715"; 
            journalName: "Nat Biotechnol"; date: "2016-12"; volume: "34"; issue:
            "12"; pages: "1279-1286"
COMMENT     Sequence Label: pLenti-sgRNA-lib 2.0
FEATURES             Location/Qualifiers
     source          1..10053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        131..510
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        512..710
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             728..908
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    955..1080
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1577..1810
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1995..2039
                     /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"
     misc_feature    2337..2454
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2534..2782
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2534..2554
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2705..2724
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     2806..2825
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2806..2824
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2854..2978
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3407..3712)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(3509..3528)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4032..4715)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     4630..4649
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4716..4818)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(4996..5120)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     5248..5264
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5306..5339)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        5455..5758
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5759..5962
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      6001..6010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6007..6009,6010..6012,6013..6726)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6007..6009,6010..6012,6013..6723)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6052..6073)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6313..6332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6660..6681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6746..6779)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6835..7423
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6888..6908)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7306..7317)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7426..7442)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7952..8132
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8194..8782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8274..8293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8953..9744,9745..9813))
                     /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     9576..9595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9814..9918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9993..10012)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
       61 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      121 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      181 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      241 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      301 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      361 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      421 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      481 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      541 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      601 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      661 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
      721 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
      781 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
      841 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
      901 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
      961 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     1021 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     1081 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     1141 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     1201 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     1261 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     1321 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     1381 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     1441 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1501 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1561 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1621 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1681 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1741 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1801 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1861 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1921 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1981 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     2041 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     2101 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     2161 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     2221 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2281 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     2341 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2401 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2461 tattacaggg acagcagaga tccagtttgg ttagtaccgg gcccgctcta gaggtaccaa
     2521 ggtctggcag gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
     2581 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
     2641 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
     2701 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
     2761 cttgtggaaa ggacgaaaca ccgagagacg ctggcttatc gaaattaata cgactcacta
     2821 tagggagacc caagctggct agttaagcta tcaacaagtt tgtacaaaaa agctgaacga
     2881 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     2941 acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt agatggtatt
     3001 agtgacctgt agtcgaccga cagccttcca aatgttcttc gggtgatgct gccaacttag
     3061 tcgaccgaca gccttccaaa tgttcttctc aaacggaatc gtcgtatcca gcctactcgc
     3121 tattgtcctc aatgccgtat taaatcataa aaagaaataa gaaaaagagg tgcgagcctc
     3181 ttttttgtgt gacaaaataa aaacatctac ctattcatat acgctagtgt catagtcctg
     3241 aaaatcatct gcatcaagaa caatttcaca actcttatac ttttctctta caagtcgttc
     3301 ggcttcatct ggattttcag cctctatact tactaaacgt gataaagttt ctgtaatttc
     3361 tactgtatcg acctgcagac tggctgtgta taagggagcc tgacatttat attccccaga
     3421 acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt
     3481 ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat
     3541 ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac
     3601 tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca
     3661 caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc atttcaccag
     3721 cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac ctcagccatc
     3781 ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc attctgcatg
     3841 gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac tgatagctgt
     3901 cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg acatacttcg
     3961 ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg
     4021 ttatctggct tttagtaagc cggatccacg cggcgtttac gccccccctg ccactcatcg
     4081 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     4141 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     4201 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     4261 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     4321 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     4381 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     4441 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
     4501 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     4561 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     4621 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     4681 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     4741 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     4801 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     4861 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
     4921 tcggcgcaaa gtgcgtcggg tgatgctgcc aacttagtcg actacaggtc actaatacca
     4981 tctaagtagt tgattcatag tgactggata tgttgtgttt tacagtatta tgtagtctgt
     5041 tttttatgca aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca
     5101 gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaacgtct ctgtttaaga
     5161 gctatgctgg aaacagcata gcaagtttaa ataaggctag tccgttatca acttgaaaaa
     5221 gtggcaccga gtcggtgctt tttttctcga ggtcgacggt atcgataagc tcgcttcacg
     5281 agattccagc aggtcgaggg acctaataac ttcgtatagc atacattata cgaagttata
     5341 ttaagggttc caagcttaag cggccgcgtg gataaccgta ttaccgccat gcattagtta
     5401 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     5461 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5521 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5581 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     5641 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     5701 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     5761 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     5821 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     5881 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     5941 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
     6001 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     6061 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     6121 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     6181 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     6241 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     6301 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     6361 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     6421 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     6481 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     6541 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     6601 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     6661 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     6721 aagtaggaat tcgtcgaggg acctaataac ttcgtatagc atacattata cgaagttata
     6781 catgtttaag ggttccggtt ccactaggta caattcgata tcaagcttat cgataatcaa
     6841 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6901 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6961 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     7021 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     7081 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     7141 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     7201 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     7261 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     7321 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     7381 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     7441 cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     7501 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     7561 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     7621 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     7681 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     7741 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     7801 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     7861 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     7921 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     7981 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     8041 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     8101 acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc
     8161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     8221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     8281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     8341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     8401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     9001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     9061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     9121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     9181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     9241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     9301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     9361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     9421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     9961 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    10021 agccagtatc tgctccctgc ttgtgtgttg gag
//
