LOCUS       sequence-237577-       12991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CasRx (NLS-RfxCas13d-NLS) with 2A-EGFP for RNA targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12991)
  AUTHORS   Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD
  TITLE     Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR 
            Effectors.
  JOURNAL   Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 
            10.1016/j.cell.2018.02.033.
  PUBMED    29551272
REFERENCE   2  (bases 1 to 12991)
  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"; journalName: "Cell. 2018 
            Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033."
COMMENT     Sequence Label: pXR001: EF1a-CasRx-2A-EGFP
FEATURES             Location/Qualifiers
     source          1..12991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    61..178
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        303..1481
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          534..1472
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1429..1449
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1500..1520
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1530..4427
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             4437..4457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4467..4493
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4509..4562
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4563..4565,4566..5279)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4563..4565,4566..5276)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4605..4626)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4866..4885)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5213..5234
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5304..5892
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5357..5377)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5775..5786)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5895..5911)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5896..5912)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6417..6597
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6623..6640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6629..6853
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6899..7327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6986..7005)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7196..7217
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7336..7356)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7341..7670
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7521..7656
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7583..7602
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7718..7765
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7784..8158
                     /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    8288..8409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8325..8344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8379..8398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8458..8474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8458..8474)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8471..8493)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8482..8498
                     /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(8506..8512,8513..8530,8531..8536))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8551..8572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8689..8706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8860..9448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8940..8959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9619..10410,10411..10479))
                     /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     10242..10261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10480..10584)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10659..10678)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10850..11229
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11231..11429
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11384..11404
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11447..11627
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11674..11799
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12292..12525
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12710..12754
                     /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 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
       61 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
      121 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
      181 ggtttattac agggacagca gagatccagt ttggttaatt aatgcaaaga tggataaagt
      241 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      301 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      361 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      421 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      481 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      541 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      601 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      661 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      721 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      781 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      841 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      901 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      961 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     1021 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     1081 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     1141 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1201 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1261 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1321 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1381 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1441 cagtggttca aagttttttt cttccatttc aggtgtcgtg acgtacggcc accatgagcc
     1501 ccaagaagaa gagaaaggtg gaggccagca tcgaaaaaaa aaagtccttc gccaagggca
     1561 tgggcgtgaa gtccacactc gtgtccggct ccaaagtgta catgacaacc ttcgccgaag
     1621 gcagcgacgc caggctggaa aagatcgtgg agggcgacag catcaggagc gtgaatgagg
     1681 gcgaggcctt cagcgctgaa atggccgata aaaacgccgg ctataagatc ggcaacgcca
     1741 aattcagcca tcctaagggc tacgccgtgg tggctaacaa ccctctgtat acaggacccg
     1801 tccagcagga tatgctcggc ctgaaggaaa ctctggaaaa gaggtacttc ggcgagagcg
     1861 ctgatggcaa tgacaatatt tgtatccagg tgatccataa catcctggac attgaaaaaa
     1921 tcctcgccga atacattacc aacgccgcct acgccgtcaa caatatctcc ggcctggata
     1981 aggacattat tggattcggc aagttctcca cagtgtatac ctacgacgaa ttcaaagacc
     2041 ccgagcacca tagggccgct ttcaacaata acgataagct catcaacgcc atcaaggccc
     2101 agtatgacga gttcgacaac ttcctcgata accccagact cggctatttc ggccaggcct
     2161 ttttcagcaa ggagggcaga aattacatca tcaattacgg caacgaatgc tatgacattc
     2221 tggccctcct gagcggactg aggcactggg tggtccataa caacgaagaa gagtccagga
     2281 tctccaggac ctggctctac aacctcgata agaacctcga caacgaatac atctccaccc
     2341 tcaactacct ctacgacagg atcaccaatg agctgaccaa ctccttctcc aagaactccg
     2401 ccgccaacgt gaactatatt gccgaaactc tgggaatcaa ccctgccgaa ttcgccgaac
     2461 aatatttcag attcagcatt atgaaagagc agaaaaacct cggattcaat atcaccaagc
     2521 tcagggaagt gatgctggac aggaaggata tgtccgagat caggaaaaat cataaggtgt
     2581 tcgactccat caggaccaag gtctacacca tgatggactt tgtgatttat aggtattaca
     2641 tcgaagagga tgccaaggtg gctgccgcca ataagtccct ccccgataat gagaagtccc
     2701 tgagcgagaa ggatatcttt gtgattaacc tgaggggctc cttcaacgac gaccagaagg
     2761 atgccctcta ctacgatgaa gctaatagaa tttggagaaa gctcgaaaat atcatgcaca
     2821 acatcaagga atttagggga aacaagacaa gagagtataa gaagaaggac gcccctagac
     2881 tgcccagaat cctgcccgct ggccgtgatg tttccgcctt cagcaaactc atgtatgccc
     2941 tgaccatgtt cctggatggc aaggagatca acgacctcct gaccaccctg attaataaat
     3001 tcgataacat ccagagcttc ctgaaggtga tgcctctcat cggagtcaac gctaagttcg
     3061 tggaggaata cgcctttttc aaagactccg ccaagatcgc cgatgagctg aggctgatca
     3121 agtccttcgc tagaatggga gaacctattg ccgatgccag gagggccatg tatatcgacg
     3181 ccatccgtat tttaggaacc aacctgtcct atgatgagct caaggccctc gccgacacct
     3241 tttccctgga cgagaacgga aacaagctca agaaaggcaa gcacggcatg agaaatttca
     3301 ttattaataa cgtgatcagc aataaaaggt tccactacct gatcagatac ggtgatcctg
     3361 cccacctcca tgagatcgcc aaaaacgagg ccgtggtgaa gttcgtgctc ggcaggatcg
     3421 ctgacatcca gaaaaaacag ggccagaacg gcaagaacca gatcgacagg tactacgaaa
     3481 cttgtatcgg aaaggataag ggcaagagcg tgagcgaaaa ggtggacgct ctcacaaaga
     3541 tcatcaccgg aatgaactac gaccaattcg acaagaaaag gagcgtcatt gaggacaccg
     3601 gcagggaaaa cgccgagagg gagaagttta aaaagatcat cagcctgtac ctcaccgtga
     3661 tctaccacat cctcaagaat attgtcaata tcaacgccag gtacgtcatc ggattccatt
     3721 gcgtcgagcg tgatgctcaa ctgtacaagg agaaaggcta cgacatcaat ctcaagaaac
     3781 tggaagagaa gggattcagc tccgtcacca agctctgcgc tggcattgat gaaactgccc
     3841 ccgataagag aaaggacgtg gaaaaggaga tggctgaaag agccaaggag agcattgaca
     3901 gcctcgagag cgccaacccc aagctgtatg ccaattacat caaatacagc gacgagaaga
     3961 aagccgagga gttcaccagg cagattaaca gggagaaggc caaaaccgcc ctgaacgcct
     4021 acctgaggaa caccaagtgg aatgtgatca tcagggagga cctcctgaga attgacaaca
     4081 agacatgtac cctgttcaga aacaaggccg tccacctgga agtggccagg tatgtccacg
     4141 cctatatcaa cgacattgcc gaggtcaatt cctacttcca actgtaccat tacatcatgc
     4201 agagaattat catgaatgag aggtacgaga aaagcagcgg aaaggtgtcc gagtacttcg
     4261 acgctgtgaa tgacgagaag aagtacaacg ataggctcct gaaactgctg tgtgtgcctt
     4321 tcggctactg tatccccagg tttaagaacc tgagcatcga ggccctgttc gataggaacg
     4381 aggccgccaa gttcgacaag gagaaaaaga aggtgtccgg caattccgga tccggaccta
     4441 agaaaaagag gaaggtggcg gccgcttacc catacgatgt tccagattac gctgctagcg
     4501 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     4561 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     4621 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     4681 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     4741 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     4801 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     4861 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     4921 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     4981 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     5041 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     5101 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     5161 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     5221 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     5281 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     5341 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5401 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5461 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5521 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5581 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     5641 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     5701 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     5761 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     5821 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     5881 ccgcctcccc gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa
     5941 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     6001 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     6061 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6121 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     6181 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     6241 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
     6301 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
     6361 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
     6421 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6481 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6541 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
     6601 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6661 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6721 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6781 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     6841 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     6901 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6961 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7021 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7081 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7141 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7201 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7261 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7321 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     7381 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     7441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     7501 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     7561 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     7621 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     7681 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     7741 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     7801 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     7861 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     7921 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     7981 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     8041 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     8101 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     8161 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     8221 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     8281 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8401 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     8461 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     8521 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     8581 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     8641 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     8701 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     8761 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8821 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8881 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     8941 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     9001 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     9061 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     9121 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     9181 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     9241 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     9301 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     9361 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     9421 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     9481 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     9541 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     9601 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     9661 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     9721 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     9781 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     9841 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     9901 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     9961 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    10021 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    10081 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    10141 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    10201 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    10261 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    10321 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    10381 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    10441 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    10501 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    10561 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
    10621 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
    10681 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
    10741 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
    10801 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
    10861 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
    10921 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
    10981 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
    11041 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
    11101 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
    11161 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
    11221 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
    11281 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
    11341 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
    11401 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
    11461 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
    11521 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
    11581 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
    11641 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
    11701 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
    11761 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
    11821 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
    11881 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
    11941 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
    12001 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
    12061 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
    12121 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
    12181 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
    12241 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
    12301 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
    12361 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
    12421 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
    12481 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
    12541 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
    12601 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
    12661 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
    12721 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
    12781 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
    12841 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
    12901 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
    12961 agagagagac agagacagat ccattcgatt a
//
