LOCUS       sequence-332228-       11980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of EsCas13d in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11980)
  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 11980)
  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-EsCas13d-msfGFP-NES-Flag
FEATURES             Location/Qualifiers
     source          1..11980
                     /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(4097..4099,4100..4810)
                     /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="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     4747..4768
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4820..4846
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             4871..4936
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4943..5510
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5104..5121)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5331..5350
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5530..6129
                     /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(5530..5549)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6026..6046
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6148..6736
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6201..6221)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6619..6630)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6811..7044
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7163..7181)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7249..7353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7354..7422,7423..8214)
                     /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(7572..7591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8385..8973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8874..8893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9127..9144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9384..9405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9420..9425,9426..9443,9444..9450)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9458..9474
                     /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     9463..9485
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9482..9498
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9482..9498
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9548..9567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9759..10392
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10439..10564
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11061..11294
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11479..11523
                     /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    11816..11933
                     /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 ccgctagcgg caagaagatc cacgcccggg acctgagaga
     1261 gcagaggaag accgaccgga cagagaagtt cgccgatcag aacaagaaga gggaggcaga
     1321 gagggcagtg cccaagaagg acgccgccgt gagcgtgaag tccgtgagct ccgtgtctag
     1381 caagaaggat aatgtgacca agtctatggc caaggcagca ggagtgaaga gcgtgttcgc
     1441 agtgggcaac accgtgtaca tgacatcctt tggcagaggc aatgacgccg tgctggagca
     1501 gaagatcgtg gatacatctc acgagccact gaacatcgac gatcccgcct accagctgaa
     1561 tgtggtgacc atgaacggct attccgtgac aggacacagg ggagagaccg tgtctgccgt
     1621 gacagacaac ccactgcgga gattcaatgg ccgcaagaag gacgagcccg agcagtctgt
     1681 gcctaccgat atgctgtgcc tgaagcccac actggagaag aagttctttg gcaaggagtt
     1741 tgacgataac atccacatcc agctgatcta caatatcctg gacatcgaga agatcctggc
     1801 cgtgtacagc accaacgcca tctatgccct gaacaatatg agcgccgacg agaatatcga
     1861 gaactccgat ttctttatga agaggaccac agatgagacc ttcgacgatt ttgagaagaa
     1921 gaaggagtct acaaacagcc gggagaaggc cgacttcgat gcctttgaga agttcatcgg
     1981 caattacaga ctggcctatt ttgccgacgc cttctacgtg aataagaaga accccaaggg
     2041 caaggccaag aatgtgctgc gcgaggataa ggagctgtat tctgtgctga ccctgatcgg
     2101 caagctgagg cactggtgcg tgcacagcga ggagggaaga gcagagttct ggctgtacaa
     2161 gctggacgag ctgaaggacg acttcaagaa cgtgctggat gtggtgtata ataggcctgt
     2221 ggaggagatc aacaatcgct tcatcgagaa caataaggtg aacatccaga tcctgggcag
     2281 cgtgtacaag aataccgaca tcgccgagct ggtgagatcc tactatgagt tcctgatcac
     2341 aaagaagtat aagaacatgg gcttttccat caagaagctg cgggagtcta tgctggaggg
     2401 caagggctac gccgacaagg agtatgattc tgtgagaaat aagctgtacc agatgaccga
     2461 cttcatcctg tacacaggct atatcaacga ggacagcgat agggccgacg atctggtgaa
     2521 taccctgcgc tcctctctga aggaggacga taagaccaca gtgtattgca aggaggccga
     2581 ctacctgtgg aagaagtatc gggagtccat cagagaggtg gccgatgccc tggacggcga
     2641 taacatcaag aagctgagca agtccaatat cgagatccag gaggacaagc tgaggaagtg
     2701 ttttatcagc tacgccgatt ctgtgagcga gttcaccaag ctgatctatc tgctgacacg
     2761 ctttctgtcc ggcaaggaga tcaacgacct ggtgaccaca ctgatcaata agtttgataa
     2821 catcaggtct ttcctggaga tcatggacga gctgggcctg gatcgcacct tcacagccga
     2881 gtacagcttc tttgagggct ccaccaagta tctggccgag ctggtggagc tgaacagctt
     2941 tgtgaagtcc tgctctttcg acatcaatgc caagaggacc atgtaccgcg acgccctgga
     3001 tatcctgggc atcgagtccg acaagacaga ggaggatatc gagaagatga tcgacaacat
     3061 cctgcagatc gacgccaatg gcgataagaa gctgaagaag aacaatggcc tgcggaattt
     3121 catcgcctct aacgtgatcg atagcaatcg gtttaagtac ctggtgagat atggcaaccc
     3181 taagaagatc agggagaccg ccaagtgcaa gcctgccgtg cgcttcgtgc tgaatgagat
     3241 cccagatgcc cagatcgagc ggtactatga ggcctgctgt ccaaagaaca ccgccctgtg
     3301 cagcgccaat aagaggcgcg agaagctggc cgacatgatc gccgagatca agtttgagaa
     3361 cttctccgat gccggcaact accagaaggc caatgtgacc tccagaacat ctgaggccga
     3421 gatcaagagg aagaaccagg ccatcatccg cctgtacctg accgtgatgt atatcatgct
     3481 gaagaatctg gtgaacgtga atgccagata cgtgatcgcc tttcactgcg tggagagaga
     3541 cacaaagctg tatgccgaga gcggcctgga ggtcggcaac atcgagaaga ataagaccaa
     3601 cctgacaatg gccgtgatgg gcgtgaagct ggagaatggc atcatcaaga ccgagtttga
     3661 caagtccttc gccgagaatg ccgccaaccg ctacctgcgg aacgccagat ggtataagct
     3721 gatcctggat aatctgaaga agtctgagcg ggccgtggtg aatgagttca gaaacaccgt
     3781 gtgccacctg aacgccatca ggaacatcaa tatcaacatc aaggagatca aggaggtgga
     3841 gaactacttt gccctgtacc actatctgat ccagaagcac ctggagaatc ggttcgccga
     3901 caagaaggtg gagagagaca ccggcgactt catctccaag ctggaggagc acaagaccta
     3961 ctgcaaggac tttgtgaagg cctattgtac acccttcggc tacaacctgg tgcgctataa
     4021 gaatctgaca atcgacggcc tgttcgataa gaactaccct ggcaaggacg atagcgacga
     4081 gcagaagagt ctagaggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     4141 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg
     4201 cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     4261 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     4321 cgaccacatg aagcgccacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     4381 gcgcaccatc agcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga
     4441 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4501 catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata tcaccgccga
     4561 caagcagaag aacggcatca aggccaactt caagatccgc cacaacgtgg aggacggcag
     4621 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4681 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
     4741 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactcacg gcatggacga
     4801 gctgtacaag taccttcaac tgcctccact tgaaagactg acactgctag agcaacgcgt
     4861 gcagctaagc gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa
     4921 ggatgacgat gacaagtaag gatccctccc ccccccctaa cgttactggc cgaagccgct
     4981 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     5041 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     5101 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     5161 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     5221 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     5281 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     5341 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     5401 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     5461 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa ggatccaccg
     5521 gagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5581 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5641 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5701 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5761 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5821 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5881 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5941 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     6001 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     6061 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     6121 ggtgcctgac gcccatcgat agatcctaat caacctctgg attacaaaat ttgtgaaaga
     6181 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6241 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6301 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6361 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6421 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6481 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6541 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6601 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6661 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6721 tgggccgcct ccccgcctga gatcctttaa gaccaatgac ttacaaggca gctgtagatc
     6781 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     6841 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6901 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6961 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     7021 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     7081 aacttgcaaa gaaatgaata tcagagagtg agaggcccgg gttaattaag gaaagggcta
     7141 gatcattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     7201 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7261 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7321 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7381 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7441 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7501 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7561 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     7621 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7681 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7741 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     7801 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7861 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     7921 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7981 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8041 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     8101 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8161 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8221 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8281 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8341 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8401 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8461 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8521 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     8581 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8641 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8701 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8761 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8821 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8881 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8941 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     9001 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     9061 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9121 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9181 cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg ctgactaatt ttttttattt
     9241 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     9301 ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa
     9361 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     9421 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     9481 acaggaaaca gctatgacat gattacgaat ttcacaaata aagcattttt ttcactgcat
     9541 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata
     9601 actcaagcta accaaaatca tcccaaactt cccaccccat accctattac cactgccaat
     9661 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
     9721 aattgtattt gttaaatatg tactacaaac ttagtagttg gaagggctaa ttcactccca
     9781 aagaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta
     9841 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa
     9901 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt
     9961 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag
    10021 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa
    10081 gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg
    10141 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt
    10201 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
    10261 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
    10321 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
    10381 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
    10441 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
    10501 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
    10561 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
    10621 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
    10681 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
    10741 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
    10801 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
    10861 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc gctgatcttc
    10921 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
    10981 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
    11041 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
    11101 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
    11161 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
    11221 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
    11281 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
    11341 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
    11401 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
    11461 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
    11521 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
    11581 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
    11641 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
    11701 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
    11761 gtgaacggat ctcgacggta tcgccgaatt cacaaatggc agtattcatc cacaatttta
    11821 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
    11881 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
    11941 attacaggga cagcagagat ccagtttgga ctagtcgtga
//
