LOCUS       sequence-437114-       10325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The backbone lentiviral dual-CRISPR plasmid for making the 
            dual-CRISPR screening libraries..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10325)
  AUTHORS   Li Y, Tan M, Akkari-Henic A, Zhang L, Kip M, Sun S, Sepers JJ, Xu N,
            Ariyurek Y, Kloet SL, Davis RP, Mikkers H, Gruber JJ, Snyder MP, Li 
            X, Pang B
  TITLE     Genome-wide Cas9-mediated screening of essential non-coding 
            regulatory elements via libraries of paired single-guide RNAs.
  JOURNAL   Nat Biomed Eng. 2024 May 22. doi: 10.1038/s41551-024-01204-8.
  PUBMED    38778183
REFERENCE   2  (bases 1 to 10325)
  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: "Nat Biomed 
            Eng. 2024 May 22. doi: 10.1038/s41551-024-01204-8."
COMMENT     Sequence Label: Lenti-dual-CRISPR-U6-H1(pBP43)
FEATURES             Location/Qualifiers
     source          1..10325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(194..238)
                     /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    423..656
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1149..1274
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1321..1501
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1502..1728)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(1756..1776)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1756..1774)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1795..1811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1795..1811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1808..1830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1819..1835
                     /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)."
     promoter        complement(join(1843..1849,1850..1867,1868..1873))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1888..1909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2026..2043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2197..2785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2277..2296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2956..3747,3748..3816))
                     /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     3579..3598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3817..3921)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3947..4402)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4084..4105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4296..4315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4529..4551
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4543..4560
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4544..4560
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4570..4589
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4570..4588
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      4609..4744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4784..4905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4795..4814)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4849..4868
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             4983..5216
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    5288..5876
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5394..5405
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5803..5823
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5892..6491)
                     /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(5975..5995)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6472..6491
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6504..7682)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6513..7451
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6536..6556)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    7821..7938
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7988..8202
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     8123..8142
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        complement(join(10093..10325,1..16))
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(10151..10170)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(join(10321..10325,1..16))
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tgggaaatag gccctctctg ggtcccctcg gggttgggag gtgggtctga aacgataatg
       61 gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact attcttaaac
      121 ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa tttgttatgt
      181 taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt tcattctttt
      241 cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt
      301 ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat ttattactcc
      361 aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc
      421 ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc tggagctgct
      481 tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat
      541 tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc agcgtcattg
      601 acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa gctcctattc
      661 ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt gctactccta
      721 atggttcaat ttttactact ttatatttat ataattcact tctccaattg tccctcatat
      781 ctcctcctcc aggtctgaag atcagcggcc gcttgctgtg cggtggtctt acttttgttt
      841 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
      901 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
      961 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
     1021 ggattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
     1081 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
     1141 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
     1201 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
     1261 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
     1321 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     1381 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     1441 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     1501 cgtttatgta tcgagctagg cacttaaata caatatctct gcaatgcggc aattcagtgg
     1561 ttcgtccaat ccatgtcaga cccgtctgtt gccttcctaa taaggcacga tcgtaccacc
     1621 ttacttccac caatcggcat gcacggtgct ttttctctcc ttgtaaggca tgttgctaac
     1681 tcatcgttac catgttgcaa gactacaaga gtattgcata agactacatt aagcttgcag
     1741 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     1801 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1861 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1921 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1981 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2041 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2101 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2161 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2221 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2281 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2341 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2401 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2461 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2521 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2581 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2641 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2701 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2821 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2881 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2941 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3001 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3061 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3121 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3181 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3241 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3301 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3361 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3421 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3481 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3541 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3601 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3661 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3721 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3781 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3841 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3901 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     3961 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4021 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4081 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4141 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4201 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4261 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4321 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4381 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4441 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     4501 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     4561 gagcgcgcgt aatacgactc actatagggc gaattgggta cgtccctagg cctccaaaaa
     4621 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     4681 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     4741 ggatgggcgg agttaggggc gggatagcta gagccagaca tgataagata cattgatgag
     4801 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4861 gctattgctt tatttgtaac cattataagc tgcaataaac aagttcctct cactctctga
     4921 tattcatttc tttgcaagtt ataaatactg aataataaga tgacatgaac tactatacgt
     4981 actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     5041 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     5101 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     5161 cccagtacaa gcaaaaagca gatcttgtct tcgttgggag tgaattagcc cttccagtcc
     5221 ccccttttct tttaaaaagt ggctaagatc tacagctgcc ttgtaagtca ttggtcttaa
     5281 agtcgacgcg gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga
     5341 cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg
     5401 gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc
     5461 agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc
     5521 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg
     5581 agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg
     5641 ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac
     5701 tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     5761 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     5821 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattgtcg
     5881 acttaacgcg ttcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     5941 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     6001 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     6061 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agactccgac ggtggccagg
     6121 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     6181 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     6241 gcttcgacgc tctccggcgt ggtccagacc gccacggcgg cgccgtcgtc cgcgacccac
     6301 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     6361 tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     6421 gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     6481 tactcggtca tggtggccgt acgtcacgac acctgaaatg gaagaaaaaa actttgaacc
     6541 actgtctgag gcttgagaat gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag
     6601 aattacatca agtgccaagc tggcctaact tcagtctcca cccactcagt gtggggaaac
     6661 tccatcgcat aaaacccctc cccccaacct aaagacgacg tactccaaaa gctcgagaac
     6721 taatcgaggt gcctggacgg cgcccggtac tccgtggagt cacatgaagc gacggctgag
     6781 gacggaaagg cccttttcct ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc
     6841 gtcctccatt ttgagctccc tgcagcaggg ccgggaagcg gccatctttc cgctcacgca
     6901 actggtgccg accgggccag ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg
     6961 ccaggcacca gagcaggccg gccagcttga gactaccccc gtccgattct cggtggccgc
     7021 gctcgcaggc cccgcctcgc cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc
     7081 gcggccccaa aaaccgaaat accagtgtgc agatcttggc ccgcatttac aagactatct
     7141 tgccagaaaa aaagcgtcgc agcaggtcat caaaaatttt aaatggctag agacttatcg
     7201 aaagcagcga gacaggcgcg aaggtgccac cagattcgca cgcggcggcc ccagcgccca
     7261 ggccaggcct caactcaagc acgaggcgaa ggggctcctt aagcgcaagg cctcgaactc
     7321 tcccacccac ttccaacccg aagctcggga tcaagaatca cgtactgcag ccaggtggaa
     7381 gtaattcaag gcacgcaagg gccataaccc gtaaagaggc caggcccgcg ggaaccacac
     7441 acggcactta cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact
     7501 actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca
     7561 tcactttccc agtttacccc gcgccacctt ctctaggcac cggatcaatt gccgacccct
     7621 ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgacg ggcaccggag
     7681 ccaattccca ctcctttcaa gacctagaag gtccattagc tgcaaagatt cctctctgtt
     7741 taaaacttta tccatctttg cacccgggcc ccctcgagcc ggcgccaaag tggatctctg
     7801 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
     7861 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
     7921 cccccttttc ttttaaaatt gtggatgaat actgccattt gtctcaagat ctagttacgc
     7981 caagcttgaa cgctgacgtc atcaacccgc tccaaggaat cgcgggccca gtgtcactag
     8041 gcgggaacac ccagcgcgcg tgcgccctgg caggaagatg gctgtgaggg acaggggagt
     8101 ggcgccctgc aatatttgca tgtcgctatg tgttctggga aatcaccata aacgtgaaat
     8161 gtctttggat ttgggaatct tataagttct gtatgagacc actctttccc gagagacgta
     8221 caaaaaagag caagaagcta aaaaagattt aaaaattatt tttagcgcag ttaatggaac
     8281 aggaactaaa tttaccccaa aaatattacg tgaatcagga tataacgtta ttgaggttga
     8341 agagcatgca tttgaagatg aaacatttaa aaatgttgta aatccaaatc cagaatttga
     8401 tcctgcatga aaaataccgc ttgaatatgg tattaaacat gatgcagata ttattattat
     8461 gaatgaccca gatgctgaca gatttggaat ggcaataaaa catgatggtc attttgtaag
     8521 attagatgga aatcaaacag gaccaatttt aattgattga aaattatcaa atctaaaacg
     8581 cttaaatagc attccaaaaa atccggctct atattcaagt tttgtaacaa gtgatttggg
     8641 tgatagaatc gctcatgaaa aatatggagt taatattgta aaaactttaa ctggatttaa
     8701 atgaatgggt agagaaattg ctaaagaaga agataacgga ttaaattttg tttttgctta
     8761 tgaagaaagt tatggatatg taattgatga ctcagctaga gataaagatg gaatacaagc
     8821 ttctatatta atagcagagg ctgcttgatt ttataaaaaa caaaataaaa cattagtaga
     8881 ctatttagaa gatttattta aagaaatggg tgcatattac actttcactt taaacttgaa
     8941 ttttaaacca gaagaaaaga aattaaaaat tgaaccatta atgaaatcat tgagagcaac
     9001 acccttaact caaattgctg gacttaaagt tgttaatgtt gaagactaca tcgatggaat
     9061 gtataatatg ccaggacaag acttactaaa attttattta gaagataagt catgatttgc
     9121 tgttcgccca agtggaactg aacctaaact aaaaatttat tttataggtg ttggtgaatc
     9181 tgttcaaaac gctaaagtta aagtagacga aattattaaa gaattaaaat taaaaatgaa
     9241 tatataggag aaaaaatgaa actaaacaaa tatatagatc acacattatt aaaacaagat
     9301 gctacgaaag ctgaaattaa acaattatgt gatgaagcaa ttgaatttga ttttgcaaca
     9361 gtttgtgtta attcatattg aacaagctat tgtaaagaat tattaaaagg cacaaatgta
     9421 ggaataacaa atgttgtagg ttttcctcta ggtgcatgca caacagctac aaaagcattc
     9481 gaagtttctg aagcaattaa agatggtgca acagaaattg atatggtatt aaatattggt
     9541 gcattaaaag acaaaaatta tgaattagtt ttagaagaca tgaaagctgt aaaaaaagca
     9601 gctggatcac atgttgttaa atgtattatg gaaaattgtt tattaacaaa agaagaaatc
     9661 atgaaagctt gtgaaatagc tgttgaagct ggattagaat ttgttaaaac atcaacagga
     9721 ttttcaaaat caggtgcaac atttgaagat gttaaactaa tgaagtcagt tgttaaagac
     9781 aatgctttag ttaaagcagc tggtggagtt agaacatttg aagatgctca aaaaatgatt
     9841 gaagcaggag ctgaccgctt aggaacaagt ggtggagtag ctattattaa aggtgaagaa
     9901 aacaacgcga gttactaaaa ctagcgtttt tttattttgc tcatttttat taaaagtttg
     9961 caaaaaggaa cataaaaatt ctaattattg atactaaagt tattaaaaag aagattttgg
    10021 ttgattttat aaaggtcata gaatataata ttttagcatg tgtattttgt gtgctcattt
    10081 acaaccgtct ccggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata
    10141 ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa
    10201 ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac
    10261 agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg
    10321 aatca
//
