LOCUS       sequence-375155-        9699 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For cloning of guide RNAs compatible with RfxCas13d. Contains a 5' 
            direct repeat and smRNA, evopreQ1, capture sequences and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9699)
  AUTHORS   Wessels HH, Mendez-Mancilla A, Hao Y, Papalexi E, Mauck WM 3rd, Lu 
            L, Morris JA, Mimitou EP, Smibert P, Sanjana NE, Satija R
  TITLE     Efficient combinatorial targeting of RNA transcripts in single cells
            with Cas13 RNA Perturb-seq.
  JOURNAL   Nat Methods. 2023 Jan;20(1):86-94. doi: 10.1038/s41592-022-01705-x. 
            Epub 2022 Dec 22.
  PUBMED    36550277
REFERENCE   2  (bases 1 to 9699)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-022-01705-x"; journalName: "Nat Methods"; date: 
            "2023-01"; volume: "20"; issue: "1"; pages: "86-94"
COMMENT     Sequence Label: pLentiRNAGuide_003
FEATURES             Location/Qualifiers
     source          1..9699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(235..339)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(414..433)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        605..984
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        986..1184
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1139..1159
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1202..1382
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1429..1554
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2047..2280
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2465..2509
                     /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    2807..2924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2975..3223
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2975..2995
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3146..3165
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3226..3254
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d 
                     gRNA (Konermann et al., 2018)"
     misc_RNA        3271..3306
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     promoter        3484..3695
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3709..3718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3718..3759
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             join(3766..3768,3769..4479)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3808..3829)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4069..4088)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4416..4437
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4489..4545
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4546..5142
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5039..5059
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5158..5746
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5211..5231)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5629..5640)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5818..6051
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6077..6094)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6083..6307
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6353..6781
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6440..6459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6650..6671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6790..6810)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6795..7124
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6975..7110
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7037..7056
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7172..7219
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7238..7612
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7742..7863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7779..7798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7833..7852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7912..7928)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7912..7928)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7925..7947)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7936..7952
                     /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(7960..7966,7967..7984,7985..7990))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8005..8026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8143..8160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8314..8902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8394..8413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9073..9699,1..165,166..234))
                     /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     join(9696..9699,1..16)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
       61 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
      121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
      181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      361 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
      421 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
      481 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
      541 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
      601 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      661 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      721 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      781 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      841 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      901 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      961 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     1021 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     1081 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     1141 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac
     1201 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     1261 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     1321 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     1381 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     1441 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     1501 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     1561 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     1621 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     1681 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     1741 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     1801 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggtagag
     1861 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     1921 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1981 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     2041 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     2101 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     2161 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     2221 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     2281 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     2341 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     2401 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     2461 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     2521 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     2581 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2641 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2701 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2761 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2821 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2881 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2941 acagcagaga tccagtttgg ttaattaagg taccgagggc ctatttccca tgattccttc
     3001 atatttgcat atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa
     3061 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
     3121 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
     3181 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgtacccc taccaactgg
     3241 tcggggtttg aaacggagac ggacgtctca caagtaaacc cctaccaact ggtcggggtt
     3301 tgaaaccctt ggcacccgag aattccatga agagctacga tatcatgctc ttctgcttta
     3361 aggccggtcc tagcaagatc ttgacgcggt tctatctagt tacgcgttaa accaactaga
     3421 aattttttga attcgctagc taggtcttga aaggagtggg aattgactcc ggtgcccgtc
     3481 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     3541 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     3601 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     3661 ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagagc caccatgggt
     3721 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtacggtgag caagggcgag
     3781 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3841 aagttcagcg tgtctggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3901 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3961 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     4021 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4081 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4141 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4201 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggcgaacttc
     4261 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4321 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4381 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4441 gccgccggga tcactctcgg catggacgag ctgtacaagg gatccggcgc aacaaacttc
     4501 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgaccga gtacaagccc
     4561 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     4621 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     4681 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4741 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4801 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4861 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4921 accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4981 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5041 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5101 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat
     5161 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     5221 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5281 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5341 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5401 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5461 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5521 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     5581 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5641 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     5701 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     5761 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5821 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     5881 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5941 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6001 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt
     6061 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6121 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6181 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     6241 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     6301 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     6361 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6421 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6481 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6541 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6601 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6661 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6721 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6781 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     6841 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     6901 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     6961 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     7021 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     7081 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     7141 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     7201 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     7261 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     7321 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     7381 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     7441 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     7501 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     7561 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     7621 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     7681 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     7741 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7861 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7921 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7981 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     8041 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     8101 cgcggggaga ggcggtttgc gtattgggcg ctcatccgct tcctcgctca ctgactcgct
     8161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     8221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     8281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     8341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     8401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     8461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     8521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     9001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     9061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     9121 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     9181 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     9241 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     9301 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     9361 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     9421 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     9481 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     9541 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9601 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9661 gcgaccgagt tgctcttgcc cggcgtcaat acgggataa
//
