LOCUS       sequence-268920-       13898 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 13898)
  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 13898)
  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-dPspCas13b-3xEGFP-NLS-3xFlag
FEATURES             Location/Qualifiers
     source          1..13898
                     /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      6746..6755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6752..6754,6755..6757,6758..7468)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6752..6754,6755..6757,6758..7468)
                     /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(7484..7486,7487..8197)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(7484..7486,7487..8197)
                     /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(8204..8206,8207..8917)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8204..8206,8207..8917)
                     /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             8945..8992
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9011..9076
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    9083..9650
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9244..9261)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9471..9490
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9670..10269
                     /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(9670..9689)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10166..10186
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10288..10876
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10341..10361)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10759..10770)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10951..11184
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11303..11321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11389..11493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11494..11562,11563..12354)
                     /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(11712..11731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12525..13113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13014..13033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13267..13284
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13524..13545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13560..13565,13566..13583,13584..13590)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13598..13614
                     /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     13603..13625
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13622..13638
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13622..13638
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13688..13707
                     /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 cccaaaaaga agaggaaagt gaacatcccc gctctggtgg aaaaccagaa
     3481 gaagtacttt ggcacctaca gcgtgatggc catgctgaac gctcagaccg tgctggacca
     3541 catccagaag gtggccgata ttgagggcga gcagaacgag aacaacgaga atctgtggtt
     3601 tcaccccgtg atgagccacc tgtacaacgc caagaacggc tacgacaagc agcccgagaa
     3661 aaccatgttc atcatcgagc ggctgcagag ctacttccca ttcctgaaga tcatggccga
     3721 gaaccagaga gagtacagca acggcaagta caagcagaac cgcgtggaag tgaacagcaa
     3781 cgacatcttc gaggtgctga agcgcgcctt cggcgtgctg aagatgtaca gggacctgac
     3841 caacgcctac aagacctacg aggaaaagct gaacgacggc tgcgagttcc tgaccagcac
     3901 agagcaacct ctgagcggca tgatcaacaa ctactacaca gtggccctgc ggaacatgaa
     3961 cgagagatac ggctacaaga cagaggacct ggccttcatc caggacaagc ggttcaagtt
     4021 cgtgaaggac gcctacggca agaaaaagtc ccaagtgaat accggattct tcctgagcct
     4081 gcaggactac aacggcgaca cacagaagaa gctgcacctg agcggagtgg gaatcgccct
     4141 gctgatctgc ctgttcctgg acaagcagta catcaacatc tttctgagca ggctgcccat
     4201 cttctccagc tacaatgccc agagcgagga acggcggatc atcatcagat ccttcggcat
     4261 caacagcatc aagctgccca aggaccggat ccacagcgag aagtccaaca agagcgtggc
     4321 catggatatg ctcaacgaag tgaagcggtg ccccgacgag ctgttcacaa cactgtctgc
     4381 cgagaagcag tcccggttca gaatcatcag cgacgaccac aatgaagtgc tgatgaagcg
     4441 gagcagcgac agattcgtgc ctctgctgct gcagtatatc gattacggca agctgttcga
     4501 ccacatcagg ttccacgtga acatgggcaa gctgagatac ctgctgaagg ccgacaagac
     4561 ctgcatcgac ggccagacca gagtcagagt gatcgagcag cccctgaacg gcttcggcag
     4621 actggaagag gccgagacaa tgcggaagca agagaacggc accttcggca acagcggcat
     4681 ccggatcaga gacttcgaga acatgaagcg ggacgacgcc aatcctgcca actatcccta
     4741 catcgtggac acctacacac actacatcct ggaaaacaac aaggtcgaga tgtttatcaa
     4801 cgacaaagag gacagcgccc cactgctgcc cgtgatcgag gatgatagat acgtggtcaa
     4861 gacaatcccc agctgccgga tgagcaccct ggaaattcca gccatggcct tccacatgtt
     4921 tctgttcggc agcaagaaaa ccgagaagct gatcgtggac gtgcacaacc ggtacaagag
     4981 actgttccag gccatgcaga aagaagaagt gaccgccgag aatatcgcca gcttcggaat
     5041 cgccgagagc gacctgcctc agaagatcct ggatctgatc agcggcaatg cccacggcaa
     5101 ggatgtggac gccttcatca gactgaccgt ggacgacatg ctgaccgaca ccgagcggag
     5161 aatcaagaga ttcaaggacg accggaagtc cattcggagc gccgacaaca agatgggaaa
     5221 gagaggcttc aagcagatct ccacaggcaa gctggccgac ttcctggcca aggacatcgt
     5281 gctgtttcag cccagcgtga acgatggcga gaacaagatc accggcctga actaccggat
     5341 catgcagagc gccattgccg tgtacgatag cggcgacgat tacgaggcca agcagcagtt
     5401 caagctgatg ttcgagaagg cccggctgat cggcaagggc acaacagagc ctcatccatt
     5461 tctgtacaag gtgttcgccc gcagcatccc cgccaatgcc gtcgagttct acgagcgcta
     5521 cctgatcgag cggaagttct acctgaccgg cctgtccaac gagatcaaga aaggcaacag
     5581 agtggatgtg cccttcatcc ggcgggacca gaacaagtgg aaaacacccg ccatgaagac
     5641 cctgggcaga atctacagcg aggatctgcc cgtggaactg cccagacaga tgttcgacaa
     5701 tgagatcaag tcccacctga agtccctgcc acagatggaa ggcatcgact tcaacaatgc
     5761 caacgtgacc tatctgatcg ccgagtacat gaagagagtg ctggacgacg acttccagac
     5821 cttctaccag tggaaccgca actaccggta catggacatg cttaagggcg agtacgacag
     5881 aaagggctcc ctgcagcact gcttcaccag cgtggaagag agagaaggcc tctggaaaga
     5941 gcgggcctcc agaacagagc ggtacagaaa gcaggccagc aacaagatcc gcagcaaccg
     6001 gcagatgaga aacgccagca gcgaagagat cgagacaatc ctggataagc ggctgagcaa
     6061 cagccggaac gagtaccaga aaagcgagaa agtgatccgg cgctacagag tgcaggatgc
     6121 cctgctgttt ctgctggcca aaaagaccct gaccgaactg gccgatttcg acggcgagag
     6181 gttcaaactg aaagaaatca tgcccgacgc cgagaaggga atcctgagcg agatcatgcc
     6241 catgagcttc accttcgaga aaggcggcaa gaagtacacc atcaccagcg agggcatgaa
     6301 gctgaagaac tacggcgact tctttgtgct ggctagcgac aagaggatcg gcaacctgct
     6361 ggaactcgtg ggcagcgaca tcgtgtccaa agaggatatc atggaagagt tcaacaaata
     6421 cgaccagtgc aggcccgaga tcagctccat cgtgttcaac ctggaaaagt gggccttcga
     6481 cacatacccc gagctgtctg ccagagtgga ccgggaagag aaggtggact tcaagagcat
     6541 cctgaaaatc ctgctgaaca acaagaacat caacaaagag cagagcgaca tcctgcggaa
     6601 gatccggaac gccttcgatg ccaacaatta ccccgacaaa ggcgtggtgg aaatcaaggc
     6661 cctgcctgag atcgccatga gcatcaagaa ggcctttggg gagtacgcca tcatgaagga
     6721 gggagctcta gaggcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     6781 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     6841 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6901 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     6961 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     7021 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     7081 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     7141 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     7201 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     7261 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     7321 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     7381 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     7441 cactctcggc atggacgagc tgtacaagtc cggactcaga tctgtgagca agggcgagga
     7501 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     7561 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     7621 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     7681 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     7741 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     7801 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     7861 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     7921 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     7981 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     8041 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     8101 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     8161 cgccgggatc actctcggca tggacgagct gtacaagaag cttgtgagca agggcgagga
     8221 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     8281 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     8341 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     8401 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     8461 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     8521 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     8581 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     8641 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     8701 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     8761 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     8821 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     8881 cgccgggatc actctcggca tggacgagct gtacaagtcg aattctaagc caggattcca
     8941 tatgaagcga cctgccgcca caaagaaggc tggacaggct aagaagaaga aacaacgcgt
     9001 gcagctaagc gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa
     9061 ggatgacgat gacaagtaag gatccctccc ccccccctaa cgttactggc cgaagccgct
     9121 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     9181 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     9241 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     9301 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     9361 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     9421 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     9481 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     9541 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     9601 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa ggatccaccg
     9661 gagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     9721 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     9781 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     9841 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     9901 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     9961 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
    10021 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
    10081 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
    10141 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
    10201 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
    10261 ggtgcctgac gcccatcgat agatcctaat caacctctgg attacaaaat ttgtgaaaga
    10321 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
    10381 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
    10441 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
    10501 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
    10561 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
    10621 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
    10681 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
    10741 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
    10801 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
    10861 tgggccgcct ccccgcctga gatcctttaa gaccaatgac ttacaaggca gctgtagatc
    10921 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
    10981 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
    11041 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
    11101 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
    11161 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
    11221 aacttgcaaa gaaatgaata tcagagagtg agaggcccgg gttaattaag gaaagggcta
    11281 gatcattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
    11341 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
    11401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
    11461 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
    11521 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
    11581 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
    11641 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
    11701 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
    11761 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
    11821 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
    11881 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
    11941 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
    12001 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
    12061 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
    12121 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
    12181 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
    12241 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
    12301 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    12361 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    12421 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    12481 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    12541 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    12601 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    12661 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    12721 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    12781 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    12841 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    12901 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    12961 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    13021 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    13081 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    13141 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    13201 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    13261 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    13321 cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg ctgactaatt ttttttattt
    13381 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
    13441 ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc cgactggaaa
    13501 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
    13561 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
    13621 acaggaaaca gctatgacat gattacgaat ttcacaaata aagcattttt ttcactgcat
    13681 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata
    13741 actcaagcta accaaaatca tcccaaactt cccaccccat accctattac cactgccaat
    13801 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
    13861 aattgtattt gttaaatatg tactacaaac ttagtagt
//
