LOCUS       sequence-268923-       12645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12645)
  AUTHORS   Yang LZ, Wang Y, Li SQ, Yao RW, Luan PF, Wu H, Carmichael GG, Chen 
            LL
  TITLE     Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 Systems.
  JOURNAL   Mol Cell. 2019 Dec 19;76(6):981-997.e7. doi: 
            10.1016/j.molcel.2019.10.024. Epub 2019 Nov 19.
  PUBMED    31757757
REFERENCE   2  (bases 1 to 12645)
  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: 
            "10.1016/j.molcel.2019.10.024"; journalName: "Mol Cell"; date: 
            "2019-12-19- 19"; volume: "76"; issue: "6"; pages: "981-997.e7"
COMMENT     Sequence Label: pHAGE-IRES-puro-NLS-dLwaCas13a-EGFP-NLS-3xFlag
FEATURES             Location/Qualifiers
     source          1..12645
                     /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             3414..3461
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3468..6920
                     /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
                     EDFFANIDEAISSIAHGIVAFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLKI
                     FKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFWS
                     VPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQK
                     FENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIES
                     NNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKYT
                     ENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDFE
                     LEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEKI
                     ADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAIG
                     NIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEEI
                     FNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIANYI
                     AAFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKKI
                     EIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
     regulatory      6942..6951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6948..6950,6951..6953,6954..7664)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6948..6950,6951..6953,6954..7664)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6993..7014)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7254..7273)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7601..7622
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7692..7739
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7758..7823
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    7830..8397
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7991..8008)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8218..8237
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8417..9016
                     /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(8417..8436)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8913..8933
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9035..9623
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9088..9108)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9506..9517)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9698..9931
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10050..10068)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10136..10240
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10241..10309,10310..11101)
                     /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(10459..10478)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11272..11860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11761..11780
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12014..12031
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12271..12292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12307..12312,12313..12330,12331..12337)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12345..12361
                     /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     12350..12372
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12369..12385
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12369..12385
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12435..12454
                     /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 aagcggccgc atgaagcgac
     3421 ctgccgccac aaagaaggct ggacaggcta agaagaagaa agctagcaaa gtgaccaagg
     3481 tcgacggcat cagccacaag aagtacatcg aagagggcaa gctcgtgaag tccaccagcg
     3541 aggaaaaccg gaccagcgag agactgagcg agctgctgag catccggctg gacatctaca
     3601 tcaagaaccc cgacaacgcc tccgaggaag agaaccggat cagaagagag aacctgaaga
     3661 agttctttag caacaaggtg ctgcacctga aggacagcgt gctgtatctg aagaaccgga
     3721 aagaaaagaa cgccgtgcag gacaagaact atagcgaaga ggacatcagc gagtacgacc
     3781 tgaaaaacaa gaacagcttc tccgtgctga agaagatcct gctgaacgag gacgtgaact
     3841 ctgaggaact ggaaatcttt cggaaggacg tggaagccaa gctgaacaag atcaacagcc
     3901 tgaagtacag cttcgaagag aacaaggcca actaccagaa gatcaacgag aacaacgtgg
     3961 aaaaagtggg cggcaagagc aagcggaaca tcatctacga ctactacaga gagagcgcca
     4021 agcgcaacga ctacatcaac aacgtgcagg aagccttcga caagctgtat aagaaagagg
     4081 atatcgagaa actgtttttc ctgatcgaga acagcaagaa gcacgagaag tacaagatcc
     4141 gcgagtacta tcacaagatc atcggccgga agaacgacaa agagaacttc gccaagatta
     4201 tctacgaaga gatccagaac gtgaacaaca tcaaagagct gattgagaag atccccgaca
     4261 tgtctgagct gaagaaaagc caggtgttct acaagtacta cctggacaaa gaggaactga
     4321 acgacaagaa tattaagtac gccttctgcc acttcgtgga aatcgagatg tcccagctgc
     4381 tgaaaaacta cgtgtacaag cggctgagca acatcagcaa cgataagatc aagcggatct
     4441 tcgagtacca gaatctgaaa aagctgatcg aaaacaaact gctgaacaag ctggacacct
     4501 acgtgcggaa ctgcggcaag tacaactact atctgcaagt gggcgagatc gccacctccg
     4561 actttatcgc ccggaaccgg cagaacgagg ccttcctgag aaacatcatc ggcgtgtcca
     4621 gcgtggccta cttcagcctg aggaacatcc tggaaaccga gaacgagaac gatatcaccg
     4681 gccggatgcg gggcaagacc gtgaagaaca acaagggcga agagaaatac gtgtccggcg
     4741 aggtggacaa gatctacaat gagaacaagc agaacgaagt gaaagaaaat ctgaagatgt
     4801 tctacagcta cgacttcaac atggacaaca agaacgagat cgaggacttc ttcgccaaca
     4861 tcgacgaggc catcagcagc atcgcccacg gcatcgtggc cttcaacctg gaactggaag
     4921 gcaaggacat cttcgccttc aagaatatcg cccccagcga gatctccaag aagatgtttc
     4981 agaacgaaat caacgaaaag aagctgaagc tgaaaatctt caagcagctg aacagcgcca
     5041 acgtgttcaa ctactacgag aaggatgtga tcatcaagta cctgaagaat accaagttca
     5101 acttcgtgaa caaaaacatc cccttcgtgc ccagcttcac caagctgtac aacaagattg
     5161 aggacctgcg gaataccctg aagttttttt ggagcgtgcc caaggacaaa gaagagaagg
     5221 acgcccagat ctacctgctg aagaatatct actacggcga gttcctgaac aagttcgtga
     5281 aaaactccaa ggtgttcttt aagatcacca atgaagtgat caagattaac aagcagcgga
     5341 accagaaaac cggccactac aagtatcaga agttcgagaa catcgagaaa accgtgcccg
     5401 tggaatacct ggccatcatc cagagcagag agatgatcaa caaccaggac aaagaggaaa
     5461 agaataccta catcgacttt attcagcaga ttttcctgaa gggcttcatc gactacctga
     5521 acaagaacaa tctgaagtat atcgagagca acaacaacaa tgacaacaac gacatcttct
     5581 ccaagatcaa gatcaaaaag gataacaaag agaagtacga caagatcctg aagaactatg
     5641 agaagcacaa tcggaacaaa gaaatccctc acgagatcaa tgagttcgtg cgcgagatca
     5701 agctggggaa gattctgaag tacaccgaga atctgaacat gttttacctg atcctgaagc
     5761 tgctgaacca caaagagctg accaacctga agggcagcct ggaaaagtac cagtccgcca
     5821 acaaagaaga aaccttcagc gacgagctgg aactgatcaa cctgctgaac ctggacaaca
     5881 acagagtgac cgaggacttc gagctggaag ccaacgagat cggcaagttc ctggacttca
     5941 acgaaaacaa aatcaaggac cggaaagagc tgaaaaagtt cgacaccaac aagatctatt
     6001 tcgacggcga gaacatcatc aagcaccggg ccttctacaa tatcaagaaa tacggcatgc
     6061 tgaatctgct ggaaaagatc gccgataagg ccaagtataa gatcagcctg aaagaactga
     6121 aagagtacag caacaagaag aatgagattg aaaagaacta caccatgcag cagaacctgc
     6181 accggaagta cgccagaccc aagaaggacg aaaagttcaa cgacgaggac tacaaagagt
     6241 atgagaaggc catcggcaac atccagaagt acacccacct gaagaacaag gtggaattca
     6301 atgagctgaa cctgctgcag ggcctgctgc tgaagatcct gcaccggctc gtgggctaca
     6361 ccagcatctg ggagcgggac ctgagattcc ggctgaaggg cgagtttccc gagaaccact
     6421 acatcgagga aattttcaat ttcgacaact ccaagaatgt gaagtacaaa agcggccaga
     6481 tcgtggaaaa gtatatcaac ttctacaaag aactgtacaa ggacaatgtg gaaaagcgga
     6541 gcatctactc cgacaagaaa gtgaagaaac tgaagcagga aaaaaaggac ctgtacatcg
     6601 ccaactacat tgccgccttc aactacatcc cccacgccga gattagcctg ctggaagtgc
     6661 tggaaaacct gcggaagctg ctgtcctacg accggaagct gaagaacgcc atcatgaagt
     6721 ccatcgtgga cattctgaaa gaatacggct tcgtggccac cttcaagatc ggcgctgaca
     6781 agaagatcga aatccagacc ctggaatcag agaagatcgt gcacctgaag aatctgaaga
     6841 aaaagaaact gatgaccgac cggaacagcg aggaactgtg cgaactcgtg aaagtcatgt
     6901 tcgagtacaa ggccctggaa ggtctagagg cgctaccggt cgccaccatg gtgagcaagg
     6961 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     7021 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     7081 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     7141 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     7201 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     7261 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     7321 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     7381 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     7441 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     7501 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     7561 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     7621 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtcgaat tctaagccag
     7681 gattccatat gaagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaac
     7741 aacgcgtgca gctaagcgac tacaaagacc atgacggtga ttataaagat catgacatcg
     7801 attacaagga tgacgatgac aagtaaggat ccctcccccc cccctaacgt tactggccga
     7861 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     7921 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     7981 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     8041 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     8101 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     8161 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     8221 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     8281 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     8341 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagga
     8401 tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     8461 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     8521 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     8581 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     8641 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     8701 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     8761 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     8821 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     8881 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     8941 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     9001 caagcccggt gcctgacgcc catcgataga tcctaatcaa cctctggatt acaaaatttg
     9061 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     9121 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9181 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9241 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9301 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     9361 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     9421 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     9481 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     9541 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     9601 ctccctttgg gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct
     9661 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     9721 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     9781 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     9841 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     9901 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     9961 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa
    10021 agggctagat cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
    10081 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
    10141 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
    10201 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
    10261 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
    10321 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
    10381 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
    10441 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
    10501 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
    10561 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
    10621 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
    10681 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
    10741 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
    10801 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
    10861 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
    10921 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
    10981 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
    11041 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
    11101 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
    11161 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
    11221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    11281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    11341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    11401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    11461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    11521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    11581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    11641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    11701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    11761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    11821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    11881 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    11941 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    12001 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
    12061 gcctctcccc gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt
    12121 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
    12181 ggcttttttg gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga
    12241 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
    12301 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
    12361 atttcacaca ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc
    12421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
    12481 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
    12541 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt
    12601 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
