LOCUS       sequence-173711-       10054 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 10054)
  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 10054)
  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..10054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2643..2890
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2813..2832
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     2914..2933
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2914..2932
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2962..3086
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3515..3820)
                     /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(3617..3636)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4140..4823)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     4738..4757
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4824..4926)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(5104..5228)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     5357..5373
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5415..5448)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        5564..5867
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5868..6071
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      6110..6119
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6116..6118,6119..6121,6122..6835)
                     /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(6116..6118,6119..6121,6122..6832)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6161..6182)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6422..6441)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6769..6790
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6854..6887)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6943..7531
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6996..7016)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7414..7425)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7534..7550)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             8060..8240
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8302..8890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8382..8401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9061..9852,9853..9921))
                     /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     9684..9703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9922..10026)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag aggtaccaag gtcgggcagg
     2641 aagagggcct atttccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     2701 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     2761 aaagtaataa tttcttgggt atttgcagtt tttaaaatta tgttttaaaa tggactatca
     2821 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     2881 acgaaacacc gagagacgct ggcttatcga aattaatacg actcactata gggagaccca
     2941 agctggctag ttaagctatc aacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat
     3001 gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt
     3061 aaaacacaac atatccagtc actatgaatc aactacttag atggtattag tgacctgtag
     3121 tcgaccgaca gccttccaaa tgttcttcgg gtgatgctgc caacttagtc gaccgacagc
     3181 cttccaaatg ttcttctcaa acggaatcgt cgtatccagc ctactcgcta ttgtcctcaa
     3241 tgccgtatta aatcataaaa agaaataaga aaaagaggtg cgagcctctt ttttgtgtga
     3301 caaaataaaa acatctacct attcatatac gctagtgtca tagtcctgaa aatcatctgc
     3361 atcaagaaca atttcacaac tcttatactt ttctcttaca agtcgttcgg cttcatctgg
     3421 attttcagcc tctatactta ctaaacgtga taaagtttct gtaatttcta ctgtatcgac
     3481 ctgcagactg gctgtgtata agggagcctg acatttatat tccccagaac atcaggttaa
     3541 tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg
     3601 agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca
     3661 ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg gccaggggga
     3721 tcaccatccg tcgcccgggc gtgtcaataa tatcactctg tacatccaca aacagacgat
     3781 aacggctctc tcttttatag gtgtaaacct taaactgcat ttcaccagcc cctgttctcg
     3841 tcagcaaaag agccgttcat ttcaataaac cgggcgacct cagccatccc ttcctgattt
     3901 tccgctttcc agcgttcggc acgcagacga cgggcttcat tctgcatggt tgtgcttacc
     3961 agaccggaga tattgacatc atatatgcct tgagcaactg atagctgtcg ctgtcaactg
     4021 tcactgtaat acgctgcttc atagcatacc tctttttgac atacttcggg tatacatatc
     4081 agtatatatt cttataccgc aaaaatcagc gcgcaaatac gcatactgtt atctggcttt
     4141 tagtaagccg gatccacgcg gcgtttacgc cccccctgcc actcatcgca gtactgttgt
     4201 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     4261 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     4321 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     4381 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     4441 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     4501 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     4561 acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca
     4621 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     4681 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     4741 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     4801 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     4861 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     4921 cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca
     4981 ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt
     5041 gcgtcgggtg atgctgccaa cttagtcgac tacaggtcac taataccatc taagtagttg
     5101 attcatagtg actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa
     5161 atctaattta atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac
     5221 aaagtggttg atctagaggg cccgcggttc gaacgtctct gtttaagagc tatgctggaa
     5281 acagcatagc aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     5341 cggtgctttt ttttctcgag gtcgacggta tcgataagct cgcttcacga gattccagca
     5401 ggtcgaggga cctaataact tcgtatagca tacattatac gaagttatat taagggttcc
     5461 aagcttaagc ggccgcgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     5521 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     5581 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     5641 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     5701 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     5761 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     5821 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     5881 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     5941 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     6001 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     6061 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     6121 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     6181 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     6241 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     6301 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     6361 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     6421 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     6481 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     6541 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     6601 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     6661 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     6721 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     6781 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtagaattc
     6841 gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca tgtttaaggg
     6901 ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac
     6961 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     7021 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     7081 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     7141 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     7201 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     7261 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     7321 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     7381 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     7441 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     7501 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagacct
     7561 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg
     7621 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag
     7681 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     7741 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac
     7801 acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact
     7861 gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa
     7921 tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     7981 gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga
     8041 gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     8101 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     8161 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     8221 agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     8281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     8341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     8401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     8461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     8521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8701 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     8761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8821 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8881 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8941 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     9001 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     9061 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     9121 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     9181 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     9241 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     9301 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     9361 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     9421 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     9481 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     9541 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     9601 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     9661 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     9721 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9781 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9841 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9901 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9961 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
    10021 tccgcgcaca tttccccgaa aagtgccacc tgac
//
