LOCUS       sequence-332231-       12214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of RfxCas13d in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12214)
  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 12214)
  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-RfxCas13d-msfGFP-NES-Flag
FEATURES             Location/Qualifiers
     source          1..12214
                     /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             1229..4126
                     /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             join(4136..4138,4139..4849)
                     /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     complement(4178..4199)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4786..4807
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4859..4885
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             4910..4975
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4982..5549
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5143..5160)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5370..5389
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5569..6363
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5569..6363
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5623..5642)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6233..6252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    6382..6970
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6435..6455)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6853..6864)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7045..7278
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7397..7415)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7483..7587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7588..7656,7657..8448)
                     /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(7806..7825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8619..9207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9108..9127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9361..9378
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9618..9639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9654..9659,9660..9677,9678..9684)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9692..9708
                     /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     9697..9719
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9716..9732
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9716..9732
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9782..9801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9993..10626
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10673..10798
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11295..11528
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11713..11757
                     /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    12050..12167
                     /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 ccgctagcat cgagaagaag aagagcttcg ccaagggcat
     1261 gggcgtgaag agcaccctgg tgtccggctc taaggtgtac atgaccacat ttgccgaggg
     1321 ctccgacgca cggctggaga agatcgtgga gggcgatagc atcagatccg tgaacgaggg
     1381 agaggccttc agcgccgaga tggccgacaa gaatgccggc tataagatcg gcaacgccaa
     1441 gttttcccac ccaaagggct atgccgtggt ggccaacaat ccactgtaca ccggaccagt
     1501 gcagcaggac atgctgggcc tgaaggagac actggagaag agatacttcg gcgagagcgc
     1561 cgacggcaat gataacatct gcatccaggt catccacaat atcctggata tcgagaagat
     1621 cctggccgag tatatcacca acgccgccta cgccgtgaac aatatctccg gcctggacaa
     1681 ggatatcatc ggcttcggca agttttctac cgtgtacaca tatgacgagt tcaaggaccc
     1741 cgagcaccac agggccgcct ttaacaataa cgacaagctg atcaatgcca tcaaggccca
     1801 gtacgacgag ttcgataatt ttctggataa cccccggctg ggctattttg gccaggcctt
     1861 cttttctaag gagggcagaa actatatcat caattacggc aacgagtgtt acgacatcct
     1921 ggccctgctg agcggactga ggcactgggt ggtgcacaat aacgaggagg agtctaggat
     1981 cagccgcacc tggctgtata atctggacaa gaatctggat aacgagtaca tctccacact
     2041 gaactacctg tatgatcgga tcaccaatga gctgacaaac agcttctcca agaattctgc
     2101 cgccaatgtg aactacatcg cagagaccct gggcatcaac ccagcagagt tcgcagagca
     2161 gtacttccgg ttcagcatca tgaaggagca gaagaatctg ggctttaaca tcacaaagct
     2221 gcgggaagtg atgctggaca gaaaggatat gtccgagatc cggaagaacc acaaggtgtt
     2281 cgattctatc agaaccaagg tgtatacaat gatggacttt gtgatctacc ggtactatat
     2341 cgaggaggat gccaaggtgg ccgccgccaa taagagcctg cccgacaacg agaagtctct
     2401 gagcgagaag gatatcttcg tgatcaatct gagaggctcc tttaacgacg atcagaagga
     2461 cgccctgtac tatgatgagg ccaatcggat ctggagaaag ctggagaata tcatgcacaa
     2521 catcaaggag ttcaggggca acaagacccg cgagtacaag aagaaggacg caccaaggct
     2581 gcctcgcatc ctgcctgcag gaagggacgt gagcgccttc agcaagctga tgtatgccct
     2641 gacaatgttt ctggacggca aggagatcaa tgatctgctg accacactga tcaataagtt
     2701 cgacaacatc cagtcctttc tgaaagtgat gcctctgatc ggcgtgaacg ccaagttcgt
     2761 ggaggagtac gccttcttta aggacagcgc caagatcgcc gatgagctga ggctgatcaa
     2821 gtcctttgcc aggatgggag agccaatcgc agacgcaagg agagccatgt acatcgatgc
     2881 catcagaatc ctgggcacca atctgtctta tgatgagctg aaggccctgg ccgacacctt
     2941 cagcctggac gagaatggca acaagctgaa gaagggcaag cacggcatgc gcaatttcat
     3001 catcaataac gtgatcagca acaagaggtt tcactatctg atcagatacg gcgaccctgc
     3061 acacctgcac gagatcgcca agaacgaggc cgtggtgaag ttcgtgctgg gcaggatcgc
     3121 cgatatccag aagaagcagg gccagaatgg caagaaccag atcgaccgct actatgagac
     3181 ctgcatcggc aaggataagg gcaagtccgt gtctgagaag gtggacgccc tgaccaagat
     3241 catcacaggc atgaattatg accagttcga taagaagcgg agcgtgatcg aggacaccgg
     3301 aagggagaac gcagagagag agaagtttaa gaagatcatc agcctgtatc tgacagtgat
     3361 ctaccacatc ctgaagaata tcgtgaatat caacgccaga tacgtgatcg gcttccactg
     3421 cgtggagcgc gatgcccagc tgtataagga gaagggctac gacatcaacc tgaagaagct
     3481 ggaggagaag ggctttagct ccgtgaccaa gctgtgcgca ggaatcgacg agacagcacc
     3541 cgacaagcgg aaggatgtgg agaaggagat ggccgagaga gccaaggaga gcatcgactc
     3601 cctggagtct gccaatccta agctgtacgc caactatatc aagtactccg atgagaagaa
     3661 ggccgaggag ttcacccggc agatcaatag agagaaggcc aagaccgccc tgaacgccta
     3721 cctgcggaat acaaagtgga acgtgatcat cagggaggac ctgctgcgca tcgataataa
     3781 gacctgtaca ctgttcagga acaaggccgt gcacctggag gtggcccgct atgtgcacgc
     3841 ctacatcaat gacatcgccg aggtgaactc ctactttcag ctgtaccact atatcatgca
     3901 gcggatcatc atgaacgaga gatatgagaa gtctagcggc aaggtgtctg agtacttcga
     3961 cgccgtgaat gatgagaaga agtataacga tagactgctg aagctgctgt gcgtgccttt
     4021 cggctactgt atcccaaggt ttaagaatct gagcatcgag gccctgttcg acaggaacga
     4081 ggcagccaag tttgataagg agaagaagaa ggtgagcggc aactccagtc tagaggtgag
     4141 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4201 aaacggccac aagttcagcg tgcgcggcga gggcgagggc gatgccacca acggcaagct
     4261 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4321 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcgccacga
     4381 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatca gcttcaagga
     4441 cgacggcacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4501 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4561 gtacaacttc aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     4621 ggccaacttc aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta
     4681 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4741 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4801 gttcgtgacc gccgccggga tcactcacgg catggacgag ctgtacaagt accttcaact
     4861 gcctccactt gaaagactga cactgctaga gcaacgcgtg cagctaagcg actacaaaga
     4921 ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg acaagtaagg
     4981 atccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     5041 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     5101 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     5161 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     5221 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     5281 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     5341 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     5401 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     5461 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     5521 tggttttcct ttgaaaaaca cgatgataag gatccaccgg agcttaccat gattgaacaa
     5581 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5641 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5701 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     5761 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5821 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5881 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5941 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6001 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6061 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     6121 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6181 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6241 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6301 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6361 tgacgcccat cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact
     6421 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     6481 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     6541 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     6601 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     6661 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     6721 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     6781 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     6841 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     6901 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     6961 gcctccccgc ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     7021 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     7081 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     7141 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     7201 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     7261 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     7321 caaagaaatg aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat
     7381 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     7441 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     7501 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     7561 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     7621 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     7681 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     7741 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     7801 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     7861 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     7921 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     7981 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     8041 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     8101 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     8161 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     8221 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     8281 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     8341 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     8401 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     8461 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     8521 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     8581 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     8641 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     8701 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     8761 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     8821 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     8881 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     8941 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     9001 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     9061 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     9121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     9181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     9241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     9301 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     9361 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     9421 cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag
     9481 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     9541 gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc
     9601 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     9661 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     9721 aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt
     9781 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
     9841 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
     9901 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
     9961 atttgttaaa tatgtactac aaacttagta gttggaaggg ctaattcact cccaaagaag
    10021 acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa
    10081 ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt
    10141 accagttgag ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca
    10201 ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga
    10261 cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg
    10321 ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct
    10381 gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct
    10441 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
    10501 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
    10561 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
    10621 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
    10681 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
    10741 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
    10801 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
    10861 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
    10921 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
    10981 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
    11041 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
    11101 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct
    11161 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
    11221 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
    11281 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
    11341 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
    11401 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
    11461 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
    11521 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
    11581 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
    11641 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
    11701 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
    11761 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
    11821 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
    11881 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
    11941 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
    12001 ggatctcgac ggtatcgccg aattcacaaa tggcagtatt catccacaat tttaaaagaa
    12061 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
    12121 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
    12181 gggacagcag agatccagtt tggactagtc gtga
//
