LOCUS       sequence-408441-       12640 bp    DNA     circular SYN 14-APR-2025
DEFINITION  overexpression of PguCas13b in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12640)
  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 12640)
  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/s41592-020-01011-4"; journalName: "Nat Methods"; date: 
            "2021-01"; volume: "18"; issue: "1"; pages: "51-59"
COMMENT     Sequence Label: p23-NES-PguCas13b-msfGFP-NES-Flag
FEATURES             Location/Qualifiers
     source          1..12640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    163..280
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        328..1506
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          559..1497
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1454..1474
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(5081..5083,5084..5086,5087..5797)
                     /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="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(5126..5147)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     5734..5755
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5807..5833
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5858..5923
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5930..6497
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6091..6108)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6318..6337
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6517..7116
                     /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(6517..6536)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7013..7033
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7135..7723
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7188..7208)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7606..7617)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7798..8031
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8150..8168)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8236..8340
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8341..8409,8410..9201)
                     /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(8559..8578)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9372..9960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9861..9880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10114..10131
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10371..10392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10407..10412,10413..10430,10431..10437)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10445..10461
                     /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     10450..10472
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10469..10485
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10469..10485
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10535..10554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10746..11379
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11426..11551
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12048..12281
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12466..12510
                     /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 aacggatctc
      121 gacggtatcg ccgaattcac aaatggcagt attcatccac aattttaaaa gaaaaggggg
      181 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
      241 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
      301 cagagatcca gtttggacta gtcgtgaggc tccggtgccc gtcagtgggc agagcgcaca
      361 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
      421 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
      481 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
      541 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
      601 ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat
      661 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct
      721 tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg
      781 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
      841 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg
      901 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
      961 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
     1021 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
     1081 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
     1141 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
     1201 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
     1261 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta
     1321 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
     1381 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
     1441 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
     1501 tcgtgaagcg gccatgctgt accctgagcg gctgcggcgg atcctgaccg ctagcacaga
     1561 gcagagcgag aggccctaca acggcaccta ctacaccctg gaagataagc acttctgggc
     1621 cgcctttctg aacctggcca gacacaacgc ctacatcacc ctgacacaca tcgaccggca
     1681 gctggcctac agcaaggccg acatcaccaa cgaccaggac gtgctgagct tcaaggccct
     1741 gtggaagaac ttcgacaacg acctggaaag aaagagccgg ctgcggagcc tgatcctgaa
     1801 gcacttcagc tttctggaag gcgccgctta cggcaagaag ctgttcgagt ctaagagcag
     1861 cggcaacaag agcagcaaga acaaagagct gaccaagaaa gagaaagagg aactgcaggc
     1921 caacgctctg agcctggaca acctgaagtc tatcctgttc gacttcctgc agaagctgaa
     1981 ggacttccgg aactactaca gccactaccg gcacagcgga agcagcgagc tgcctctgtt
     2041 cgacggcaac atgctgcagc ggctgtacaa cgtgttcgac gtgtccgtgc agcgcgtgaa
     2101 gatcgaccac gagcacaacg atgaggtgga ccctcactac cacttcaacc acctcgtgcg
     2161 gaagggcaag aaggacagat acggccacaa cgacaacccc agcttcaagc accacttcgt
     2221 ggacggcgaa ggcatggtta cagaagccgg cctgctgttc ttcgtgtccc tgttcctgga
     2281 aaagcgggac gccatctgga tgcagaagaa gatccggggc ttcaaaggcg gcaccgagac
     2341 ataccagcag atgaccaacg aggtgttctg ccggtccaga atcagcctgc ctaagctgaa
     2401 gctggaaagc ctgcggatgg acgactggat gctgctggac atgctgaacg aactcgtgcg
     2461 gtgccccaag cctctgtacg acagactgag agaggacgac cgggcctgct ttagagtgcc
     2521 cgtggatatc ctgcctgacg aggacgatac agatggcggc ggagaggacc ccttcaagaa
     2581 taccctcgtc cggcaccagg acagattccc ttacttcgcc ctgcggtact tcgacctgaa
     2641 gaaggtgttc accagcctgc ggttccacat cgatctgggc acctaccact ttgccatcta
     2701 caagaagatg atcggcgagc agcccgagga cagacacctg accagaaacc tgtacggctt
     2761 cggccggatc caggacttcg ccgaagaaca cagacccgag gaatggaagc ggctcgtcag
     2821 agatctggac tacttcgaga caggcgacaa gccctacatc agccagacaa gcccacacta
     2881 ccacatcgag aagggaaaga tcggcctgag attcatgccc gagggccagc atctttggcc
     2941 ctctccagaa gtgggcacca ccagaaccgg cagatctaag tacgcccagg acaagagact
     3001 gaccgccgag gcctttctgt ctgtgcacga gctgatgcct atgatgttct actacttcct
     3061 gctgcgcgag aagtacagcg aggaagtgtc tgccgagaga gtgcagggca gaatcaagcg
     3121 cgtgatcgag gatgtgtacg ccgtgtacga tgcctttgcc agggacgaga tcaacacccg
     3181 ggatgagctg gatgcctgcc tggccgataa gggcatcaga agaggccatc tgcctcgcca
     3241 gatgatcgcc atcctgagcc aagagcacaa ggacatggaa gagaagattc ggaagaaact
     3301 gcaagagatg atggccgaca ccgaccaccg gctggatatg ctggatagac agaccgaccg
     3361 gaagatcaga atcggccgga agaatgccgg actgcctaag agcggagtga tcgccgattg
     3421 gctcgtgcgg gacatgatga gatttcagcc cgtggccaag gacgccagcg gaaagcctct
     3481 gaacaactcc aaggccaaca gcaccgagta ccggatgctg cagagagccc tggctctgtt
     3541 tggaggcgag aaagagaggc tgaccccata cttccggcag atgaatctga ccggcggaaa
     3601 caaccctcat ccttttctgc acgaaactcg ctgggagagc cacaccaaca tcctgtcctt
     3661 ttaccggtcc tacctgagag cccgcaaggc cttcctcgaa aggatcggca gatccgacag
     3721 agtggaaaac cggccatttc tgctgctgaa agagcccaag accgacagac agacactggt
     3781 ggctggatgg aagggcgagt tccatctgcc aagaggcatc ttcacagagg ccgtgcggga
     3841 ttgcctgatc gagatgggac acgatgaagt ggccagctac aaagaagtgg gattcatggc
     3901 caaggccgtg ccactgtact ttgagagagc ctgcgaggac agggtgcagc ccttttacga
     3961 cagccccttc aacgtgggca acagcctgaa gcctaagaag ggcagattcc tgagcaaaga
     4021 ggaacgggcc gaagagtggg agcggggcaa agagagattc cgggatctcg aagcctggtc
     4081 ctatagcgcc gccagaagaa tcgaggacgc ctttgccggc atcgagtatg ctagccccgg
     4141 aaacaagaag aaaatcgagc agctcctgcg ggacctgagc ctgtgggaag cctttgagag
     4201 caagctgaaa gtgcgggccg accggatcaa tctggccaag ctgaagaaag aaatcctgga
     4261 agcccaagaa cacccctacc acgacttcaa gagctggcag aagttcgagc gcgagctgcg
     4321 gctggtcaag aaccaggata tcatcacctg gatgatgtgc cgcgacctga tggaagaaaa
     4381 caaggtggaa ggcctggaca ccggcacact gtacctgaag gacatcagac ccaacgtgca
     4441 agagcagggc agcctgaacg tgctgaacag agtgaagccc atgagactgc ccgtggtggt
     4501 gtacagagcc gatagcagag gccacgtgca caaagaagaa gcccctctgg ccaccgtgta
     4561 catcgaggaa agagacacca agctgctgaa gcagggcaac ttcaagtcct tcgtgaagga
     4621 caggcggctg aacggcctgt tctccttcgt tgatactggc ggactggcca tggaacagta
     4681 ccccatctcc aagctgagag tggaatacga gctggccaag taccagaccg ccagagtgtg
     4741 tgtgttcgag ctgaccctga gactggaaga gtccctgctg acccggtatc ctcatctgcc
     4801 cgacgagagc ttcagagaga tgctggaatc ttggagcgac cctctgctgg ccaaatggcc
     4861 tgaactgcac ggaaaagtgc ggctgctgat cgccgtgcgg aatgccttta gccacaatca
     4921 gtaccctatg tacgacgagg ccgtgttcag ctccatccgg aagtacgatc ctagcagccc
     4981 cgacgccatc gaagagagaa tgggcctgaa tatcgcccac cgcctgtctg aggaagtgaa
     5041 gcaggccaaa gaaaccgtgg aacggatcat tcaggccagt atggtgagca agggcgagga
     5101 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     5161 gttcagcgtg cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     5221 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     5281 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cgccacgact tcttcaagtc
     5341 cgccatgccc gaaggctacg tccaggagcg caccatcagc ttcaaggacg acggcaccta
     5401 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     5461 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     5521 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     5581 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     5641 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     5701 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     5761 cgccgggatc actcacggca tggacgagct gtacaagtac cttcaactgc ctccacttga
     5821 aagactgaca ctgctagagc aacgcgtgca gctaagcgac tacaaagacc atgacggtga
     5881 ttataaagat catgacatcg attacaagga tgacgatgac aagtaaggat ccctcccccc
     5941 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     6001 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     6061 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     6121 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     6181 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     6241 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     6301 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     6361 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     6421 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     6481 gaaaaacacg atgataagga tccaccggag cttaccatga ccgagtacaa gcccacggtg
     6541 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     6601 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     6661 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     6721 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     6781 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     6841 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     6901 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     6961 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     7021 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     7081 cgcacctggt gcatgacccg caagcccggt gcctgacgcc catcgataga tcctaatcaa
     7141 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     7201 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     7261 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     7321 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     7381 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     7441 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     7501 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     7561 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     7621 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     7681 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac
     7741 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     7801 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     7861 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7921 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7981 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     8041 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     8101 ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata
     8161 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     8221 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     8281 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     8341 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     8401 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     8461 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     8521 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     8581 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     8641 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     8701 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8761 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     8821 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     8881 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     8941 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     9001 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     9061 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     9121 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     9181 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     9241 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     9301 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     9361 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     9421 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     9481 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     9541 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     9601 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     9661 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9721 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9781 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     9841 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     9901 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     9961 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
    10021 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
    10081 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
    10141 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag
    10201 ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
    10261 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg
    10321 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
    10381 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
    10441 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc
    10501 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    10561 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
    10621 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
    10681 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
    10741 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta
    10801 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
    10861 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
    10921 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
    10981 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
    11041 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
    11101 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
    11161 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
    11221 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
    11281 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
    11341 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
    11401 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
    11461 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
    11521 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
    11581 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
    11641 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
    11701 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
    11761 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
    11821 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
    11881 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
    11941 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
    12001 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
    12061 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
    12121 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
    12181 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
    12241 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
    12301 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
    12361 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
    12421 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
    12481 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
    12541 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
    12601 gctgtacttt ctatagtgaa tagagttagg cagggatatt
//
