LOCUS       sequence-332219-       12574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of LwaCas13a in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12574)
  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 12574)
  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-LwaCas13a-msfGFP-NES-Flag
FEATURES             Location/Qualifiers
     source          1..12574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2223..3401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3349..3369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3451..6903
                     /codon_start=1
                     /product="Leptotrichia wadei CRISPR-Cas effector that acts 
                     as an RNA-guided RNAse (Abudayyeh et al., 2017)"
                     /label=LwCas13a
                     /note="previously called C2c2"
                     /translation="KVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIYI
                     KNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEYD
                     LKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINENN
                     VEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEKY
                     KIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLDK
                     EELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLLN
                     KLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETEN
                     ENDITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNEI
                     EDFFANIDEAISSIRHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLKI
                     FKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFWS
                     VPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQK
                     FENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIES
                     NNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKYT
                     ENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDFE
                     LEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEKI
                     ADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAIG
                     NIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEEI
                     FNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIRNYI
                     AHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKKI
                     EIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
     CDS             join(6913..6915,6916..7626)
                     /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(6955..6976)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     7563..7584
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7636..7662
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             7687..7752
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    7759..8326
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7920..7937)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8147..8166
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8346..8945
                     /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(8346..8365)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8842..8862
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8964..9552
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9017..9037)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9435..9446)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9627..9860
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9979..9997)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10065..10169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10170..10238,10239..11030)
                     /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(10388..10407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11201..11789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11690..11709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11943..11960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12200..12221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12236..12241,12242..12259,12260..12266)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12274..12290
                     /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     12279..12301
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12298..12314
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12298..12314
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12364..12383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccat gctgtaccct
     3421 gagcggctgc ggcggatcct gaccgctagc aaagtgacca aggtcgacgg catcagccac
     3481 aagaagtaca tcgaagaggg caagctcgtg aagtccacca gcgaggaaaa ccggaccagc
     3541 gagagactga gcgagctgct gagcatccgg ctggacatct acatcaagaa ccccgacaac
     3601 gcctccgagg aagagaaccg gatcagaaga gagaacctga agaagttctt tagcaacaag
     3661 gtgctgcacc tgaaggacag cgtgctgtat ctgaagaacc ggaaagaaaa gaacgccgtg
     3721 caggacaaga actatagcga agaggacatc agcgagtacg acctgaaaaa caagaacagc
     3781 ttctccgtgc tgaagaagat cctgctgaac gaggacgtga actctgagga actggaaatc
     3841 tttcggaagg acgtggaagc caagctgaac aagatcaaca gcctgaagta cagcttcgaa
     3901 gagaacaagg ccaactacca gaagatcaac gagaacaacg tggaaaaagt gggcggcaag
     3961 agcaagcgga acatcatcta cgactactac agagagagcg ccaagcgcaa cgactacatc
     4021 aacaacgtgc aggaagcctt cgacaagctg tataagaaag aggatatcga gaaactgttt
     4081 ttcctgatcg agaacagcaa gaagcacgag aagtacaaga tccgcgagta ctatcacaag
     4141 atcatcggcc ggaagaacga caaagagaac ttcgccaaga ttatctacga agagatccag
     4201 aacgtgaaca acatcaaaga gctgattgag aagatccccg acatgtctga gctgaagaaa
     4261 agccaggtgt tctacaagta ctacctggac aaagaggaac tgaacgacaa gaatattaag
     4321 tacgccttct gccacttcgt ggaaatcgag atgtcccagc tgctgaaaaa ctacgtgtac
     4381 aagcggctga gcaacatcag caacgataag atcaagcgga tcttcgagta ccagaatctg
     4441 aaaaagctga tcgaaaacaa actgctgaac aagctggaca cctacgtgcg gaactgcggc
     4501 aagtacaact actatctgca agtgggcgag atcgccacct ccgactttat cgcccggaac
     4561 cggcagaacg aggccttcct gagaaacatc atcggcgtgt ccagcgtggc ctacttcagc
     4621 ctgaggaaca tcctggaaac cgagaacgag aacgatatca ccggccggat gcggggcaag
     4681 accgtgaaga acaacaaggg cgaagagaaa tacgtgtccg gcgaggtgga caagatctac
     4741 aatgagaaca agcagaacga agtgaaagaa aatctgaaga tgttctacag ctacgacttc
     4801 aacatggaca acaagaacga gatcgaggac ttcttcgcca acatcgacga ggccatcagc
     4861 agcatcagac acggcatcgt gcacttcaac ctggaactgg aaggcaagga catcttcgcc
     4921 ttcaagaata tcgcccccag cgagatctcc aagaagatgt ttcagaacga aatcaacgaa
     4981 aagaagctga agctgaaaat cttcaagcag ctgaacagcg ccaacgtgtt caactactac
     5041 gagaaggatg tgatcatcaa gtacctgaag aataccaagt tcaacttcgt gaacaaaaac
     5101 atccccttcg tgcccagctt caccaagctg tacaacaaga ttgaggacct gcggaatacc
     5161 ctgaagtttt tttggagcgt gcccaaggac aaagaagaga aggacgccca gatctacctg
     5221 ctgaagaata tctactacgg cgagttcctg aacaagttcg tgaaaaactc caaggtgttc
     5281 tttaagatca ccaatgaagt gatcaagatt aacaagcagc ggaaccagaa aaccggccac
     5341 tacaagtatc agaagttcga gaacatcgag aaaaccgtgc ccgtggaata cctggccatc
     5401 atccagagca gagagatgat caacaaccag gacaaagagg aaaagaatac ctacatcgac
     5461 tttattcagc agattttcct gaagggcttc atcgactacc tgaacaagaa caatctgaag
     5521 tatatcgaga gcaacaacaa caatgacaac aacgacatct tctccaagat caagatcaaa
     5581 aaggataaca aagagaagta cgacaagatc ctgaagaact atgagaagca caatcggaac
     5641 aaagaaatcc ctcacgagat caatgagttc gtgcgcgaga tcaagctggg gaagattctg
     5701 aagtacaccg agaatctgaa catgttttac ctgatcctga agctgctgaa ccacaaagag
     5761 ctgaccaacc tgaagggcag cctggaaaag taccagtccg ccaacaaaga agaaaccttc
     5821 agcgacgagc tggaactgat caacctgctg aacctggaca acaacagagt gaccgaggac
     5881 ttcgagctgg aagccaacga gatcggcaag ttcctggact tcaacgaaaa caaaatcaag
     5941 gaccggaaag agctgaaaaa gttcgacacc aacaagatct atttcgacgg cgagaacatc
     6001 atcaagcacc gggccttcta caatatcaag aaatacggca tgctgaatct gctggaaaag
     6061 atcgccgata aggccaagta taagatcagc ctgaaagaac tgaaagagta cagcaacaag
     6121 aagaatgaga ttgaaaagaa ctacaccatg cagcagaacc tgcaccggaa gtacgccaga
     6181 cccaagaagg acgaaaagtt caacgacgag gactacaaag agtatgagaa ggccatcggc
     6241 aacatccaga agtacaccca cctgaagaac aaggtggaat tcaatgagct gaacctgctg
     6301 cagggcctgc tgctgaagat cctgcaccgg ctcgtgggct acaccagcat ctgggagcgg
     6361 gacctgagat tccggctgaa gggcgagttt cccgagaacc actacatcga ggaaattttc
     6421 aatttcgaca actccaagaa tgtgaagtac aaaagcggcc agatcgtgga aaagtatatc
     6481 aacttctaca aagaactgta caaggacaat gtggaaaagc ggagcatcta ctccgacaag
     6541 aaagtgaaga aactgaagca ggaaaaaaag gacctgtaca tccggaacta cattgcccac
     6601 ttcaactaca tcccccacgc cgagattagc ctgctggaag tgctggaaaa cctgcggaag
     6661 ctgctgtcct acgaccggaa gctgaagaac gccatcatga agtccatcgt ggacattctg
     6721 aaagaatacg gcttcgtggc caccttcaag atcggcgctg acaagaagat cgaaatccag
     6781 accctggaat cagagaagat cgtgcacctg aagaatctga agaaaaagaa actgatgacc
     6841 gaccggaaca gcgaggaact gtgcgaactc gtgaaagtca tgttcgagta caaggccctg
     6901 gagagtctag aggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     6961 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat
     7021 gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     7081 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     7141 cacatgaagc gccacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     7201 accatcagct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc
     7261 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     7321 ctggggcaca agctggagta caacttcaac agccacaacg tctatatcac cgccgacaag
     7381 cagaagaacg gcatcaaggc caacttcaag atccgccaca acgtggagga cggcagcgtg
     7441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     7501 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     7561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcacggcat ggacgagctg
     7621 tacaagtacc ttcaactgcc tccacttgaa agactgacac tgctagagca acgcgtgcag
     7681 ctaagcgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     7741 gacgatgaca agtaaggatc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     7801 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     7861 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     7921 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     7981 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     8041 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     8101 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     8161 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     8221 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     8281 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataaggat ccaccggagc
     8341 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     8401 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     8461 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     8521 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     8581 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     8641 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     8701 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     8761 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     8821 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     8881 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     8941 cctgacgccc atcgatagat cctaatcaac ctctggatta caaaatttgt gaaagattga
     9001 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     9061 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     9121 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     9181 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     9241 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     9301 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     9361 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     9421 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     9481 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     9541 ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg tagatcttag
     9601 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     9661 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     9721 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     9781 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     9841 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     9901 tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa gggctagatc
     9961 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
    10021 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
    10081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
    10141 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
    10201 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
    10261 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
    10321 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
    10381 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
    10441 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
    10501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
    10561 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
    10621 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
    10681 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
    10741 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
    10801 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
    10861 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
    10921 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
    10981 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
    11041 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
    11101 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
    11161 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
    11221 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
    11281 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
    11341 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
    11401 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
    11461 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
    11521 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
    11581 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
    11641 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
    11701 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
    11761 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    11821 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
    11881 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
    11941 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
    12001 cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt ttatttatgc
    12061 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
    12121 aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac tggaaagcgg
    12181 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
    12241 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
    12301 gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca ctgcattcta
    12361 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc
    12421 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc
    12481 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt
    12541 gtatttgtta aatatgtact acaaacttag tagt
//
