LOCUS       sequence-429074-        9254 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector encoding RfxCas13d targeting SURF2 guide array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9254)
  AUTHORS   Tieu V, Sotillo E, Bjelajac JR, Chen C, Malipatlolla M, Guerrero JA,
            Xu P, Quinn PJ, Fisher C, Klysz D, Mackall CL, Qi LS
  TITLE     A versatile CRISPR-Cas13d platform for multiplexed transcriptomic 
            regulation and metabolic engineering in primary human T cells.
  JOURNAL   Cell. 2024 Feb 29;187(5):1278-1295.e20. doi: 
            10.1016/j.cell.2024.01.035. Epub 2024 Feb 21.
  PUBMED    38387457
REFERENCE   2  (bases 1 to 9254)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2024.01.035"; journalName: "Cell"; date: 
            "2024-02-29- 29"; volume: "187"; issue: "5"; pages: "1278-1295.e20"
COMMENT     Sequence Label: pSLQ7535 pHR (hU6-crSURF2-EFS-PuroR-WPRE)
FEATURES             Location/Qualifiers
     source          1..9254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    168..285
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        336..584
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     336..356
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     507..526
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        586..615
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d 
                     gRNA (Konermann et al., 2018)"
     misc_RNA        639..674
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        698..733
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        757..792
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        816..851
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        875..910
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        934..969
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        993..1028
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        1052..1087
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        1111..1146
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     promoter        1232..1443
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             1459..2058
                     /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(1459..1478)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1955..1975
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2074..2662
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2127..2147)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2545..2556)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2734..2967
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2993..3010)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2999..3223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3269..3697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3356..3375)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3566..3587
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3706..3726)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3711..4040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3891..4026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3953..3972
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4088..4135
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4154..4528
                     /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    4658..4779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4695..4714)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4749..4768
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4828..4844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4828..4844)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4841..4863)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4852..4868
                     /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(4876..4882,4883..4900,4901..4906))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4921..4942
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5059..5076)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5230..5818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5310..5329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5989..6780,6781..6849))
                     /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     6612..6631
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6850..6954)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7029..7048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7220..7599
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7601..7799
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7754..7774
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7817..7997
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8044..8169
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8662..8895
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9080..9124
                     /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"
ORIGIN
        1 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
       61 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
      121 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
      181 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
      241 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
      301 gacagcagag atccagtttg gttaattaag gtaccgaggg cctatttccc atgattcctt
      361 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
      421 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
      481 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
      541 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgaaccc ctaccaactg
      601 gtcggggttt gaaacgagca gttcaaaatg acccagtcca agtaaacccc taccaactgg
      661 tcggggtttg aaacagcaac agacgtaaga accagagcaa gtaaacccct accaactggt
      721 cggggtttga aacaaaccaa aggaacatga ccagcccaag taaaccccta ccaactggtc
      781 ggggtttgaa acatagttga tagtaatcca gccatcaagt aaacccctac caactggtcg
      841 gggtttgaaa cagacaattg tgtcgctgcc atcgcaagta aacccctacc aactggtcgg
      901 ggtttgaaac aataatgctg ttgttgaagg cgtcaagtaa acccctacca actggtcggg
      961 gtttgaaaca aagagagagt agcgcgagca cacaagtaaa cccctaccaa ctggtcgggg
     1021 tttgaaaccc atcatgtagg ttgccgcaca gcaagtaaac ccctaccaac tggtcggggt
     1081 ttgaaacgca gttgtgtgac acggaagcgg caagtaaacc cctaccaact ggtcggggtt
     1141 tgaaaccaga agaaaagtca gaggacacct ttttttgaat tcgctagcta ggtcttgaaa
     1201 ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     1261 gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta
     1321 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     1381 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     1441 caggaccggt tctagagcat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     1501 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     1561 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     1621 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     1681 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     1741 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     1801 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc gcccgaccac
     1861 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     1921 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     1981 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     2041 cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     2101 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2161 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2221 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2281 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2341 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2401 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2461 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2521 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2581 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2641 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     2701 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     2761 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     2821 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     2881 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     2941 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
     3001 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3061 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3121 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3181 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3241 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3301 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3361 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3421 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3481 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3541 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3601 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3661 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3961 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4021 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4081 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4141 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     4201 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     4261 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     4321 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     4381 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     4441 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     4501 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     4561 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4621 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4681 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4741 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4801 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4861 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4921 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4981 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5041 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5101 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5161 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5221 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5281 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5341 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5401 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5461 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5521 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5581 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5641 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5701 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5761 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5821 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5881 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5941 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6001 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6061 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6121 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6181 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6241 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6301 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6361 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6421 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6481 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6541 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6601 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6661 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6721 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6781 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6841 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6901 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6961 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     7021 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7081 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7141 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     7201 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     7261 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     7321 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     7381 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     7441 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     7501 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     7561 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     7621 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     7681 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     7741 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc
     7801 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     7861 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     7921 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     7981 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     8041 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     8101 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     8161 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     8221 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     8281 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     8341 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     8401 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     8461 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     8521 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     8581 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     8641 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     8701 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     8761 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     8821 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     8881 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     8941 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     9001 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     9061 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     9121 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     9181 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     9241 taggcaggga tatt
//
