LOCUS       sequence-307942-       14146 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 14146)
  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 14146)
  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-dPguCas13b-3xEGFP-NLS-3xFlag
FEATURES             Location/Qualifiers
     source          1..14146
                     /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             3431..3451
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      6995..7004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7001..7003,7004..7006,7007..7717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7001..7003,7004..7006,7007..7717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7733..7735,7736..8446)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(7733..7735,7736..8446)
                     /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(8453..8455,8456..9166)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8453..8455,8456..9166)
                     /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             9194..9241
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9260..9325
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    9332..9898
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9492..9509)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9719..9738
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9918..10517
                     /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(9918..9937)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10414..10434
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10536..11124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10589..10609)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11007..11018)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11199..11432
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11551..11569)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11637..11741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11742..11810,11811..12602)
                     /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(11960..11979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12773..13361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13262..13281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13515..13532
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13772..13793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13808..13813,13814..13831,13832..13838)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13846..13862
                     /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     13851..13873
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13870..13886
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13870..13886
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13936..13955
                     /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 accggtctgc
     3421 agctagcatg cccaagaaaa agcgcaaggt aacagagcag agcgagaggc cctacaacgg
     3481 cacctactac accctggaag ataagcactt ctgggccgcc tttctgaacc tggccagaca
     3541 caacgcctac atcaccctga cacacatcga ccggcagctg gcctacagca aggccgacat
     3601 caccaacgac caggacgtgc tgagcttcaa ggccctgtgg aagaacttcg acaacgacct
     3661 ggaaagaaag agccggctgc ggagcctgat cctgaagcac ttcagctttc tggaaggcgc
     3721 cgcttacggc aagaagctgt tcgagtctaa gagcagcggc aacaagagca gcaagaacaa
     3781 agagctgacc aagaaagaga aagaggaact gcaggccaac gctctgagcc tggacaacct
     3841 gaagtctatc ctgttcgact tcctgcagaa gctgaaggac ttccggaact actacagcgc
     3901 ctaccggcac agcggaagca gcgagctgcc tctgttcgac ggcaacatgc tgcagcggct
     3961 gtacaacgtg ttcgacgtgt ccgtgcagcg cgtgaagatc gaccacgagc acaacgatga
     4021 ggtggaccct cactaccact tcaaccacct cgtgcggaag ggcaagaagg acagatacgg
     4081 ccacaacgac aaccccagct tcaagcacca cttcgtggac ggcgaaggca tggttacaga
     4141 agccggcctg ctgttcttcg tgtccctgtt cctggaaaag cgggacgcca tctggatgca
     4201 gaagaagatc cggggcttca aaggcggcac cgagacatac cagcagatga ccaacgaggt
     4261 gttctgccgg tccagaatca gcctgcctaa gctgaagctg gaaagcctgc ggatggacga
     4321 ctggatgctg ctggacatgc tgaacgaact cgtgcggtgc cccaagcctc tgtacgacag
     4381 actgagagag gacgaccggg cctgctttag agtgcccgtg gatatcctgc ctgacgagga
     4441 cgatacagat ggcggcggag aggacccctt caagaatacc ctcgtccggc accaggacag
     4501 attcccttac ttcgccctgc ggtacttcga cctgaagaag gtgttcacca gcctgcggtt
     4561 ccacatcgat ctgggcacct accactttgc catctacaag aagatgatcg gcgagcagcc
     4621 cgaggacaga cacctgacca gaaacctgta cggcttcggc cggatccagg acttcgccga
     4681 agaacacaga cccgaggaat ggaagcggct cgtcagagat ctggactact tcgagacagg
     4741 cgacaagccc tacatcagcc agacaagccc acactaccac atcgagaagg gaaagatcgg
     4801 cctgagattc atgcccgagg gccagcatct ttggccctct ccagaagtgg gcaccaccag
     4861 aaccggcaga tctaagtacg cccaggacaa gagactgacc gccgaggcct ttctgtctgt
     4921 gcacgagctg atgcctatga tgttctacta cttcctgctg cgcgagaagt acagcgagga
     4981 agtgtctgcc gagagagtgc agggcagaat caagcgcgtg atcgaggatg tgtacgccgt
     5041 gtacgatgcc tttgccaggg acgagatcaa cacccgggat gagctggatg cctgcctggc
     5101 cgataagggc atcagaagag gccatctgcc tcgccagatg atcgccatcc tgagccaaga
     5161 gcacaaggac atggaagaga agattcggaa gaaactgcaa gagatgatgg ccgacaccga
     5221 ccaccggctg gatatgctgg atagacagac cgaccggaag atcagaatcg gccggaagaa
     5281 tgccggactg cctaagagcg gagtgatcgc cgattggctc gtgcgggaca tgatgagatt
     5341 tcagcccgtg gccaaggacg ccagcggaaa gcctctgaac aactccaagg ccaacagcac
     5401 cgagtaccgg atgctgcaga gagccctggc tctgtttgga ggcgagaaag agaggctgac
     5461 cccatacttc cggcagatga atctgaccgg cggaaacaac cctcatcctt ttctgcacga
     5521 aactcgctgg gagagccaca ccaacatcct gtccttttac cggtcctacc tgagagcccg
     5581 caaggccttc ctcgaaagga tcggcagatc cgacagagtg gaaaaccggc catttctgct
     5641 gctgaaagag cccaagaccg acagacagac actggtggct ggatggaagg gcgagttcca
     5701 tctgccaaga ggcatcttca cagaggccgt gcgggattgc ctgatcgaga tgggacacga
     5761 tgaagtggcc agctacaaag aagtgggatt catggccaag gccgtgccac tgtactttga
     5821 gagagcctgc gaggacaggg tgcagccctt ttacgacagc cccttcaacg tgggcaacag
     5881 cctgaagcct aagaagggca gattcctgag caaagaggaa cgggccgaag agtgggagcg
     5941 gggcaaagag agattccggg atctcgaagc ctggtcctat agcgccgcca gaagaatcga
     6001 ggacgccttt gccggcatcg agtatgctag ccccggaaac aagaagaaaa tcgagcagct
     6061 cctgcgggac ctgagcctgt gggaagcctt tgagagcaag ctgaaagtgc gggccgaccg
     6121 gatcaatctg gccaagctga agaaagaaat cctggaagcc caagaacacc cctaccacga
     6181 cttcaagagc tggcagaagt tcgagcgcga gctgcggctg gtcaagaacc aggatatcat
     6241 cacctggatg atgtgccgcg acctgatgga agaaaacaag gtggaaggcc tggacaccgg
     6301 cacactgtac ctgaaggaca tcagacccaa cgtgcaagag cagggcagcc tgaacgtgct
     6361 gaacagagtg aagcccatga gactgcccgt ggtggtgtac agagccgata gcagaggcca
     6421 cgtgcacaaa gaagaagccc ctctggccac cgtgtacatc gaggaaagag acaccaagct
     6481 gctgaagcag ggcaacttca agtccttcgt gaaggacagg cggctgaacg gcctgttctc
     6541 cttcgttgat actggcggac tggccatgga acagtacccc atctccaagc tgagagtgga
     6601 atacgagctg gccaagtacc agaccgccag agtgtgtgtg ttcgagctga ccctgagact
     6661 ggaagagtcc ctgctgaccc ggtatcctca tctgcccgac gagagcttca gagagatgct
     6721 ggaatcttgg agcgaccctc tgctggccaa atggcctgaa ctgcacggaa aagtgcggct
     6781 gctgatcgcc gtgcggaatg cctttagcgc caatcagtac cctatgtacg acgaggccgt
     6841 gttcagctcc atccggaagt acgatcctag cagccccgac gccatcgaag agagaatggg
     6901 cctgaatatc gcccaccgcc tgtctgagga agtgaagcag gccaaagaaa ccgtggaacg
     6961 gatcattcag gccagtctag aggcgctacc ggtcgccacc atggtgagca agggcgagga
     7021 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     7081 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     7141 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     7201 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     7261 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     7321 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     7381 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     7441 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     7501 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     7561 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     7621 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     7681 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctgtgagcaa
     7741 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     7801 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     7861 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     7921 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     7981 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     8041 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     8101 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     8161 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     8221 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     8281 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     8341 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     8401 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagaagc ttgtgagcaa
     8461 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     8521 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     8581 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     8641 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     8701 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     8761 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     8821 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     8881 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     8941 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     9001 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     9061 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     9121 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtcga attctaagcc
     9181 aggattccat atgaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa
     9241 acaacgcgtg cagctaagcg actacaaaga ccatgacggt gattataaag atcatgacat
     9301 cgattacaag gatgacgatg acaagtaagg atccctcccc ccccctaacg ttactggccg
     9361 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     9421 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     9481 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     9541 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     9601 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     9661 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     9721 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     9781 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     9841 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagg
     9901 atccaccgga gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     9961 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
    10021 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
    10081 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
    10141 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
    10201 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
    10261 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
    10321 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
    10381 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
    10441 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
    10501 gcaagcccgg tgcctgacgc ccatcgatag atcctaatca acctctggat tacaaaattt
    10561 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
    10621 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
    10681 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
    10741 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
    10801 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
    10861 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
    10921 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
    10981 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
    11041 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
    11101 tctccctttg ggccgcctcc ccgcctgaga tcctttaaga ccaatgactt acaaggcagc
    11161 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
    11221 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
    11281 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    11341 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    11401 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
    11461 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggcccgggt taattaagga
    11521 aagggctaga tcattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
    11581 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
    11641 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
    11701 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
    11761 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
    11821 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
    11881 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
    11941 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
    12001 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    12061 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
    12121 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
    12181 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
    12241 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
    12301 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
    12361 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
    12421 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
    12481 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
    12541 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
    12601 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    12661 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    12721 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    12781 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    12841 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    12901 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
    12961 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    13021 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    13081 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    13141 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    13201 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    13261 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    13321 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    13381 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
    13441 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
    13501 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
    13561 cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct gactaatttt
    13621 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
    13681 aggctttttt ggaggcctag gcttttgcaa aaagctcccc gtggcacgac aggtttcccg
    13741 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
    13801 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
    13861 aatttcacac aggaaacagc tatgacatga ttacgaattt cacaaataaa gcattttttt
    13921 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
    13981 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca
    14041 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
    14101 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
