LOCUS       sequence-332222-       12379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of PspCas13b in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12379)
  AUTHORS   Li S, Li X, Xue W, Zhang L, Yang LZ, Cao SM, Lei YN, Liu CX, Guo SK,
            Shan L, Wu M, Tao X, Zhang JL, Gao X, Zhang J, Wei J, Li J, Yang L, 
            Chen LL
  TITLE     Screening for functional circular RNAs using the CRISPR-Cas13 
            system.
  JOURNAL   Nat Methods. 2021 Jan;18(1):51-59. doi: 10.1038/s41592-020-01011-4. 
            Epub 2020 Dec 7.
  PUBMED    33288960
REFERENCE   2  (bases 1 to 12379)
  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/s41592-020-01011-4"; journalName: "Nat Methods"; date: 
            "2021-01"; volume: "18"; issue: "1"; pages: "51-59"
COMMENT     Sequence Label: p23-NES-PspCas13b-msfGFP-NES-Flag
FEATURES             Location/Qualifiers
     source          1..12379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(4490..4492,4493..4495,4496..4498,4499..5209)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MKVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKD
                     DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGI
                     KANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVL
                     LEFVTAAGITHGMDELYK"
     primer_bind     complement(4538..4559)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     5146..5167
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5219..5245
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5270..5335
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5342..5909
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5503..5520)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5730..5749
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5929..6528
                     /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(5929..5948)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6425..6445
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6547..7135
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6600..6620)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7018..7029)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7210..7443
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7562..7580)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7648..7752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7753..7821,7822..8613)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7971..7990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8784..9372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9273..9292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9526..9543
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9783..9804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9819..9824,9825..9842,9843..9849)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9857..9873
                     /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)."
     primer_bind     9862..9884
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9881..9897
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9881..9897
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9947..9966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10158..10791
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10838..10963
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11460..11693
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11878..11922
                     /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    12215..12332
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa gcggccatgc tgtaccctga
     1201 gcggctgcgg cggatcctga ccgctagcaa catccccgct ctggtggaaa accagaagaa
     1261 gtactttggc acctacagcg tgatggccat gctgaacgct cagaccgtgc tggaccacat
     1321 ccagaaggtg gccgatattg agggcgagca gaacgagaac aacgagaatc tgtggtttca
     1381 ccccgtgatg agccacctgt acaacgccaa gaacggctac gacaagcagc ccgagaaaac
     1441 catgttcatc atcgagcggc tgcagagcta cttcccattc ctgaagatca tggccgagaa
     1501 ccagagagag tacagcaacg gcaagtacaa gcagaaccgc gtggaagtga acagcaacga
     1561 catcttcgag gtgctgaagc gcgccttcgg cgtgctgaag atgtacaggg acctgaccaa
     1621 ccactacaag acctacgagg aaaagctgaa cgacggctgc gagttcctga ccagcacaga
     1681 gcaacctctg agcggcatga tcaacaacta ctacacagtg gccctgcgga acatgaacga
     1741 gagatacggc tacaagacag aggacctggc cttcatccag gacaagcggt tcaagttcgt
     1801 gaaggacgcc tacggcaaga aaaagtccca agtgaatacc ggattcttcc tgagcctgca
     1861 ggactacaac ggcgacacac agaagaagct gcacctgagc ggagtgggaa tcgccctgct
     1921 gatctgcctg ttcctggaca agcagtacat caacatcttt ctgagcaggc tgcccatctt
     1981 ctccagctac aatgcccaga gcgaggaacg gcggatcatc atcagatcct tcggcatcaa
     2041 cagcatcaag ctgcccaagg accggatcca cagcgagaag tccaacaaga gcgtggccat
     2101 ggatatgctc aacgaagtga agcggtgccc cgacgagctg ttcacaacac tgtctgccga
     2161 gaagcagtcc cggttcagaa tcatcagcga cgaccacaat gaagtgctga tgaagcggag
     2221 cagcgacaga ttcgtgcctc tgctgctgca gtatatcgat tacggcaagc tgttcgacca
     2281 catcaggttc cacgtgaaca tgggcaagct gagatacctg ctgaaggccg acaagacctg
     2341 catcgacggc cagaccagag tcagagtgat cgagcagccc ctgaacggct tcggcagact
     2401 ggaagaggcc gagacaatgc ggaagcaaga gaacggcacc ttcggcaaca gcggcatccg
     2461 gatcagagac ttcgagaaca tgaagcggga cgacgccaat cctgccaact atccctacat
     2521 cgtggacacc tacacacact acatcctgga aaacaacaag gtcgagatgt ttatcaacga
     2581 caaagaggac agcgccccac tgctgcccgt gatcgaggat gatagatacg tggtcaagac
     2641 aatccccagc tgccggatga gcaccctgga aattccagcc atggccttcc acatgtttct
     2701 gttcggcagc aagaaaaccg agaagctgat cgtggacgtg cacaaccggt acaagagact
     2761 gttccaggcc atgcagaaag aagaagtgac cgccgagaat atcgccagct tcggaatcgc
     2821 cgagagcgac ctgcctcaga agatcctgga tctgatcagc ggcaatgccc acggcaagga
     2881 tgtggacgcc ttcatcagac tgaccgtgga cgacatgctg accgacaccg agcggagaat
     2941 caagagattc aaggacgacc ggaagtccat tcggagcgcc gacaacaaga tgggaaagag
     3001 aggcttcaag cagatctcca caggcaagct ggccgacttc ctggccaagg acatcgtgct
     3061 gtttcagccc agcgtgaacg atggcgagaa caagatcacc ggcctgaact accggatcat
     3121 gcagagcgcc attgccgtgt acgatagcgg cgacgattac gaggccaagc agcagttcaa
     3181 gctgatgttc gagaaggccc ggctgatcgg caagggcaca acagagcctc atccatttct
     3241 gtacaaggtg ttcgcccgca gcatccccgc caatgccgtc gagttctacg agcgctacct
     3301 gatcgagcgg aagttctacc tgaccggcct gtccaacgag atcaagaaag gcaacagagt
     3361 ggatgtgccc ttcatccggc gggaccagaa caagtggaaa acacccgcca tgaagaccct
     3421 gggcagaatc tacagcgagg atctgcccgt ggaactgccc agacagatgt tcgacaatga
     3481 gatcaagtcc cacctgaagt ccctgccaca gatggaaggc atcgacttca acaatgccaa
     3541 cgtgacctat ctgatcgccg agtacatgaa gagagtgctg gacgacgact tccagacctt
     3601 ctaccagtgg aaccgcaact accggtacat ggacatgctt aagggcgagt acgacagaaa
     3661 gggctccctg cagcactgct tcaccagcgt ggaagagaga gaaggcctct ggaaagagcg
     3721 ggcctccaga acagagcggt acagaaagca ggccagcaac aagatccgca gcaaccggca
     3781 gatgagaaac gccagcagcg aagagatcga gacaatcctg gataagcggc tgagcaacag
     3841 ccggaacgag taccagaaaa gcgagaaagt gatccggcgc tacagagtgc aggatgccct
     3901 gctgtttctg ctggccaaaa agaccctgac cgaactggcc gatttcgacg gcgagaggtt
     3961 caaactgaaa gaaatcatgc ccgacgccga gaagggaatc ctgagcgaga tcatgcccat
     4021 gagcttcacc ttcgagaaag gcggcaagaa gtacaccatc accagcgagg gcatgaagct
     4081 gaagaactac ggcgacttct ttgtgctggc tagcgacaag aggatcggca acctgctgga
     4141 actcgtgggc agcgacatcg tgtccaaaga ggatatcatg gaagagttca acaaatacga
     4201 ccagtgcagg cccgagatca gctccatcgt gttcaacctg gaaaagtggg ccttcgacac
     4261 ataccccgag ctgtctgcca gagtggaccg ggaagagaag gtggacttca agagcatcct
     4321 gaaaatcctg ctgaacaaca agaacatcaa caaagagcag agcgacatcc tgcggaagat
     4381 ccggaacgcc ttcgatcaca acaattaccc cgacaaaggc gtggtggaaa tcaaggccct
     4441 gcctgagatc gccatgagca tcaagaaggc ctttggggag tacgccatca tgaaggtgag
     4501 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4561 aaacggccac aagttcagcg tgcgcggcga gggcgagggc gatgccacca acggcaagct
     4621 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4681 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcgccacga
     4741 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatca gcttcaagga
     4801 cgacggcacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4861 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4921 gtacaacttc aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     4981 ggccaacttc aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta
     5041 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5101 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5161 gttcgtgacc gccgccggga tcactcacgg catggacgag ctgtacaagt accttcaact
     5221 gcctccactt gaaagactga cactgctaga gcaacgcgtg cagctaagcg actacaaaga
     5281 ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg acaagtaagg
     5341 atccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     5401 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     5461 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     5521 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     5581 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     5641 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     5701 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     5761 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     5821 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     5881 tggttttcct ttgaaaaaca cgatgataag gatccaccgg agcttaccat gaccgagtac
     5941 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     6001 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     6061 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     6121 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     6181 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     6241 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     6301 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     6361 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     6421 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     6481 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccatcgata
     6541 gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     6601 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     6661 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     6721 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     6781 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     6841 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     6901 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     6961 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     7021 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     7081 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgag
     7141 atcctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     7201 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     7261 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     7321 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     7381 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7441 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     7501 cagagagtga gaggcccggg ttaattaagg aaagggctag atcattcttg aagacgaaag
     7561 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     7621 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     7681 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     7741 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     7801 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     7861 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7921 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7981 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     8041 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     8101 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     8161 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     8221 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     8281 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     8341 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     8401 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     8461 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     8521 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     8581 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     8641 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     8701 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     8761 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     8821 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     8881 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     8941 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     9001 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     9061 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     9121 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     9181 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     9241 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     9301 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     9361 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     9421 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     9481 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     9541 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     9601 taatgcagca agctcatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     9661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     9721 aaaagctccc cgtggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     9781 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     9841 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg
     9901 attacgaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9961 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
    10021 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
    10081 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
    10141 actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga
    10201 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
    10261 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
    10321 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
    10381 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
    10441 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
    10501 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
    10561 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
    10621 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
    10681 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
    10741 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
    10801 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
    10861 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
    10921 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
    10981 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
    11041 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
    11101 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
    11161 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
    11221 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
    11281 taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg
    11341 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
    11401 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
    11461 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
    11521 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
    11581 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
    11641 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
    11701 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
    11761 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
    11821 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
    11881 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
    11941 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
    12001 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
    12061 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
    12121 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
    12181 cgccgaattc acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
    12241 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
    12301 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
    12361 cagtttggac tagtcgtga
//
