pHAGE-IRES-puro-NLS-NLS-dPspCas13b-3xsfGFP-NLS

LOCUS       pHAGE-IRES-puro-NLS-NLS-dPspCas13b-3xsfGFP-NLS       13819 bp    DNA     circular SYN 14-APR-2025
DEFINITION  overexpression in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13819)
  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 13819)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.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-NLS-dPspCas13b-3xsfGFP-NLS
FEATURES             Location/Qualifiers
     source          1..13819
                     /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             3471..3491
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6777..7490
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             7509..8222
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             8241..8954
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             8961..8981
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    8998..9571
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9165..9182)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9392..9411
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9591..10190
                     /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(9591..9610)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10087..10107
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10209..10797
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10262..10282)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10680..10691)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10872..11105
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11224..11242)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11310..11414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11415..11483,11484..12275)
                     /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(11633..11652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12446..13034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12935..12954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13188..13205
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13445..13466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13481..13486,13487..13504,13505..13511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13519..13535
                     /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     13524..13546
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13543..13559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13543..13559
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13609..13628
                     /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 agctagcatg cccaaaaaga
     3481 agaggaaagt gaacatcccc gccctggtgg agaatcagaa gaagtacttc ggcacctatt
     3541 ccgtgatggc catgctgaac gcccagacag tgctggatca catccagaag gtggccgaca
     3601 tcgagggcga gcagaacgag aacaatgaga atctgtggtt tcaccctgtg atgagccacc
     3661 tgtacaacgc caagaatggc tatgataagc agcctgaaaa gaccatgttc atcatcgaga
     3721 gactgcagtc ctacttccca tttctgaaga tcatggccga gaatcagcgc gagtactcta
     3781 acggcaagta taagcagaat cgggtggagg tgaacagcaa tgacatcttc gaggtgctga
     3841 agagagcctt tggcgtgctg aagatgtaca gggatctgac caacgcctac aagacatatg
     3901 aggagaagct gaatgacggc tgcgagtttc tgaccagcac agagcagcct ctgtccggca
     3961 tgatcaacaa ttactatacc gtggccctga gaaacatgaa tgagaggtac ggctacaaga
     4021 cagaggatct ggccttcatc caggacaaga ggttcaagtt tgtgaaggac gcctacggca
     4081 agaagaagag ccaggtgaac accggcttct ttctgtctct gcaggattat aatggcgaca
     4141 cacagaagaa gctgcacctg agcggagtgg gaatcgccct gctgatctgc ctgtttctgg
     4201 ataagcagta catcaacatc ttcctgagca gactgccaat ctttagctcc tataatgccc
     4261 agtccgagga gcggagaatc atcatccgca gcttcggcat caactccatc aagctgccaa
     4321 aggacagaat ccactccgag aagtctaaca agagcgtggc catggatatg ctgaatgagg
     4381 tgaagaggtg tcccgacgag ctgttcacca cactgtctgc cgagaagcag agccggttta
     4441 gaatcatctc cgacgatcac aacgaggtgc tgatgaagag atctagcgac aggtttgtgc
     4501 ctctgctgct gcagtacatc gattatggca agctgttcga ccacatccgc tttcacgtga
     4561 acatgggcaa gctgcggtac ctgctgaagg ccgataagac ctgcatcgac ggccagacaa
     4621 gggtgcgcgt gatcgagcag ccactgaatg gattcggccg cctggaggag gcagagacca
     4681 tgcggaagca ggagaacggc acattcggca atagcggcat ccggatcaga gattttgaga
     4741 acatgaagcg ggacgatgcc aaccctgcca attacccata tatcgtggac acctacacac
     4801 actatatcct ggagaacaat aaggtggaga tgtttatcaa tgataaggag gactccgccc
     4861 cactgctgcc cgtgatcgag gacgatagat acgtggtgaa gaccatccca agctgtagga
     4921 tgtccacact ggagatcccc gccatggcct tccacatgtt cctgtttggc tctaagaaga
     4981 ccgagaagct gatcgtggac gtgcacaacc ggtataagag actgttccag gccatgcaga
     5041 aggaggaggt gacagccgag aatatcgcct cttttggcat cgccgagagc gatctgcccc
     5101 agaagatcct ggacctgatc tccggcaacg cccacggcaa ggatgtggac gccttcatcc
     5161 gcctgaccgt ggacgatatg ctgaccgata cagagaggcg catcaagcgg tttaaggacg
     5221 atcgcaagtc catccggtct gccgacaata agatgggcaa gcgcggcttc aagcagatct
     5281 ctaccggcaa gctggccgat ttcctggcca aggacatcgt gctgtttcag ccaagcgtga
     5341 acgatggcga gaataagatc acaggcctga actaccggat catgcagagc gccatcgccg
     5401 tgtacgactc cggcgacgat tatgaggcca agcagcagtt caagctgatg tttgagaagg
     5461 ccagactgat cggcaagggc accacagagc cccacccttt cctgtacaag gtgtttgcaa
     5521 ggtccatccc agcaaacgca gtggagttct acgagcggta cctgatcgag cggaagtttt
     5581 atctgaccgg cctgtctaac gagatcaaga agggcaatcg cgtggatgtg cccttcatcc
     5641 ggagagacca gaataagtgg aagacccctg ccatgaagac actgggcaga atctacagcg
     5701 aggatctgcc cgtggagctg cctaggcaga tgtttgacaa cgagatcaag tctcacctga
     5761 agagcctgcc tcagatggag ggcatcgact tcaacaatgc caatgtgacc tacctgatcg
     5821 ccgagtatat gaagcgggtg ctggacgatg acttccagac cttctaccag tggaacagga
     5881 attaccgcta tatggatatg ctgaagggcg agtatgacag aaagggctct ctgcagcact
     5941 gcttcaccag cgtggaggag agggagggac tgtggaagga gagggcatcc agaacagaga
     6001 ggtaccgcaa gcaggcctcc aacaagatca ggtctaatcg ccagatgcgg aacgcctcct
     6061 ctgaggagat cgagaccatc ctggataaga gactgtctaa cagcaggaat gagtaccaga
     6121 agagcgagaa agtgatcagg cgctatcgcg tgcaggacgc cctgctgttc ctgctggcca
     6181 agaagaccct gacagagctg gccgatttcg acggcgagcg gtttaagctg aaggagatca
     6241 tgccagatgc cgagaagggc atcctgtccg agatcatgcc catgtctttc acatttgaga
     6301 agggcggcaa gaagtacacc atcacaagcg agggcatgaa gctgaagaac tatggcgatt
     6361 tctttgtgct ggcctccgac aagagaatcg gcaatctgct ggagctggtg ggctccgata
     6421 tcgtgtctaa ggaggacatc atggaggagt ttaacaagta cgatcagtgt aggcccgaga
     6481 tcagctccat cgtgttcaat ctggagaagt gggcctttga cacctatcct gagctgtccg
     6541 ccagagtgga tagggaggag aaggtggact tcaagtctat cctgaagatc ctgctgaaca
     6601 ataagaacat caataaggag cagtctgaca tcctgcgcaa gatccggaac gcctttgatg
     6661 ccaacaatta ccctgacaag ggcgtggtgg agatcaaggc cctgccagag atcgccatga
     6721 gcatcaagaa ggccttcggc gagtatgcca tcatgaaggg aggtggatcc ggaagtatgc
     6781 gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg
     6841 tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac
     6901 tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg actctggtaa
     6961 cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg
     7021 acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg
     7081 atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc
     7141 gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg
     7201 aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta
     7261 aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact
     7321 accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga
     7381 gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg
     7441 agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa tctggcggcc
     7501 gctctggcat gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac
     7561 tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa
     7621 ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg gtaccttggc
     7681 cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata
     7741 tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga
     7801 tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata
     7861 ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaagacggc aatatcctgg
     7921 gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa
     7981 aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc
     8041 tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca
     8101 atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata
     8161 tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca
     8221 aaggtagcat cgatggtagc atgcgtaaag gcgaagagct gttcactggt gtcgtcccta
     8281 ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg
     8341 aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc
     8401 cggtaccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt
     8461 atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc
     8521 aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat
     8581 ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg
     8641 gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg
     8701 ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg
     8761 gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc
     8821 tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga
     8881 aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg
     8941 atgaactgta caaactcgag ccaaagaaaa agcggaaagt gtaatctaga ggatctgccc
     9001 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     9061 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     9121 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     9181 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     9241 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     9301 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     9361 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaatgg
     9421 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     9481 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     9541 cgtggttttc ctttgaaaaa cacgatgata aggatccacc ggagcttacc atgaccgagt
     9601 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     9661 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     9721 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     9781 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     9841 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     9901 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     9961 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
    10021 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
    10081 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
    10141 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccatcga
    10201 tagatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
    10261 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
    10321 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
    10381 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
    10441 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
    10501 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
    10561 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
    10621 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
    10681 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
    10741 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
    10801 agatccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
    10861 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
    10921 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
    10981 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
    11041 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
    11101 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
    11161 atcagagagt gagaggcccg ggttaattaa ggaaagggct agatcattct tgaagacgaa
    11221 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
    11281 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
    11341 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
    11401 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
    11461 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
    11521 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
    11581 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
    11641 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
    11701 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
    11761 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
    11821 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
    11881 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
    11941 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
    12001 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
    12061 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
    12121 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
    12181 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
    12241 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
    12301 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
    12361 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
    12421 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
    12481 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
    12541 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
    12601 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
    12661 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
    12721 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
    12781 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
    12841 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
    12901 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
    12961 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
    13021 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
    13081 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
    13141 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
    13201 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
    13261 attaatgcag caagctcatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
    13321 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
    13381 caaaaagctc cccgtggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
    13441 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
    13501 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca
    13561 tgattacgaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
    13621 aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc
    13681 atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac
    13741 tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat
    13801 gtactacaaa cttagtagt
//