LOCUS       sequence-375405-       12762 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Overexpression of NLS-dPba5Cas13b-EGFP-NLS in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12762)
  AUTHORS   Yang LZ, Gao BQ, Huang Y, Wang Y, Yang L, Chen LL
  TITLE     Multi-color RNA imaging with CRISPR-Cas13b systems in living cells.
  JOURNAL   Cell Insight. 2022 Jun 15;1(4):100044. doi: 
            10.1016/j.cellin.2022.100044. eCollection 2022 Aug.
  PUBMED    37192858
REFERENCE   2  (bases 1 to 12762)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Insight."; date: "2022-06-15"; volume: "1(4):100044. doi"; pages: " 
            10.1016/j.cellin.2022.100044. eCollection 2022 Aug"
COMMENT     Sequence Label: pHAGE-IRES-puro-NLS-dPba5Cas13b-EGFP-NLS
FEATURES             Location/Qualifiers
     source          1..12762
                     /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"
     regulatory      7059..7068
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7065..7067,7068..7070,7071..7781)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7065..7067,7068..7070,7071..7781)
                     /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(7110..7131)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7371..7390)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7718..7739
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7809..7856
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7875..7940
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    7947..8514
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8108..8125)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8335..8354
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8534..9133
                     /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(8534..8553)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9030..9050
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9152..9740
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9205..9225)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9623..9634)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9815..10048
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10167..10185)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10253..10357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10358..10426,10427..11218)
                     /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(10576..10595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11389..11977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11878..11897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12131..12148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12388..12409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12424..12429,12430..12447,12448..12454)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12462..12478
                     /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     12467..12489
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12486..12502
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12486..12502
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12552..12571
                     /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 atggtggagt
     3481 tcgataagaa tagaaagtac atctacgccg cctactgtaa catggcccag cacaacttca
     3541 tcagcaccct gaagcacatc gccaagtgtc tgtctctgga tctggagctg accgataaaa
     3601 gcaagaaaat catcgacgag ctgaaggata aggtgaacag cctgacacag gataagaagc
     3661 tgaagttcta cgaactgctg aacaagcact tccctttcct ggcccccatg attgacagcg
     3721 ataagagcgc caaggattac aacgagatca aggccatcaa tgacatcctc ccacagatcg
     3781 gcaccgtggt ggatttctac gccgactaca ccaccgccta cgaccccgac gaggagaagt
     3841 ccgtggacta cctgcatccc agcgagcaga ccctggtgcc cagactgaag gagctgctga
     3901 aggctgagat ccgcaccgtg cagcagaggt ttaactatgt ggaggatgag gtgctgttca
     3961 ttaagaacaa ggagatggac aaggagagat ataggtactg cctgtttaag aagctgtgcc
     4021 ggaccgataa gaaggtgttc tcaaggaggg gcctggtgtt tttcatcagc ctgttcctgg
     4081 agaagaaata cattaacgag atgctcaata agaccaagga cttttacgcc aaggacgacc
     4141 tgttcaacga taagcgcaaa cacattatct ttgaatctct ggccgtgtac agaattaggc
     4201 tgcccaggaa tcggtacgag agccagcagg atgacgtggg cctggccctg gacatggtga
     4261 acgagctgca gcggtgccct aaggaactgt tcgacctgct gtccaccgag gaccagatca
     4321 aattcagacc agacgcttct gatgaggact cctccaagac agccatcatg attcgccatg
     4381 ccgaccggtt ccctaagctc gccatgaact atattgatca gtgcaatgtg tttaactcca
     4441 tcaggttcca ggtgaacctg ggcaagtacc gctacgcctt ctacaacaag aagtgtattg
     4501 acggcctgag ccatgtgagg tccctgcaga aggagctgca cgggttcggg aagctgaatg
     4561 agattgagaa ggctaggctg gacacatggg gaaacctgat ccgggacttc gaggatatca
     4621 agaccaacac cattgacaca atgccctaca ttaatgacca gagagccagc tacatcatta
     4681 acaacaaccg gatcggcctg tactgggagg ataaggatga cgtgaatgag cctaagctcg
     4741 tgctgccaaa gctgcctaat gagcctaccg ccaagaacct gcggcagcag aaggccaacg
     4801 gcgagcggat cgtgctcctg cagtctccca agtgcttcat gtccacccac gagctgcctg
     4861 ctctgatctt ctaccacatc atccggaaga agcaccagaa aaccgatagc cctgacgtgg
     4921 agtccatcat tatcaaatgg gtggaggggt tccggaactt tatccaggag gtcatggatg
     4981 gcaagattac ctcccagaac gctcaggagc gggccaatca gctgggcatt aactacgccc
     5041 agcatctgcc taagaagatc caggagtacc ttgatgccaa ccaggataac aataacgcca
     5101 tcaccgccaa tctgcagaag aagttgaaga acagactgac cacccacatt gatgagacca
     5161 agaagctgat cgaaaagtac aacaacgtga agcacgtgga ctcaaagaaa aataagcgcg
     5221 gcaccaagcg gttcgtggag atcaaacagg ggcgcctggg cgcctggctg gcccaggaca
     5281 ttattgccct gcagcctgcc cctgccgacg gcaacaataa gctgaccggc ctgaactttc
     5341 agctgctgca gagcgccctg agcatctatg ataactttga caacctgaag cggctgctga
     5401 tcgccgccca cctgatccag tccaagggca atcacccctt tctcagcgag attgtgtcta
     5461 agagccctca gaccatcggg gacttctacc aggcctacct gaaggctaaa ctgaattggc
     5521 tgaactccct gctgagcgag gaggatctga ttaagtaccc ttttctgaca aggggcgtta
     5581 acaagtggaa gttcagggac gaggagtatt acaagagcat catgcagcag tacctggagc
     5641 tccccgccga gctgcccaga ggcctgtttg aaacagccat caaggacatc ctgatcgact
     5701 gcttcggccg cgggtttgtg aggggcaaca atcgcgagga ggccaacgtg gccttcctga
     5761 tcgccaagta tttcaagaag gtttttaacg atgactacca ggagttttat actcagcccg
     5821 atggcaatta caagaggcac tactacttct tcaaactgct gctgcagtcc gataaggact
     5881 acggcaagac cgtgaaggaa atcgaggaca tcctgcgcac cgacatgagc ctgaccctgc
     5941 tgagtaaaac caccatcgag aatgacgacg acagtgacaa ctctgataac agcgacctga
     6001 aggagtactt tgaggagatc aataagaagc tggccaaaag cctcgccaag gaccccactc
     6061 tggacaagag gcacaggatc gaggacatta tcagcaagag cgagaccctc tctaagctgc
     6121 caaactccca gcagaagaaa ctcctgagga ttagcagcaa gcgacacggc gtgcccatca
     6181 atgacggcct ggtgaagaaa cacatcaaga agatcacctc caactccgag aaggaggagg
     6241 agctgaagaa gctgagccac gccctgcaca agctgaagtc tacagagagg ctgatcagga
     6301 gatatagagt gcaggacatc atcattttca tgatggctag ggagatcctg ttcgagaacg
     6361 ataagaacct gtttgagaat agatttgaag agttcaagct gaagaacatc aggcctatta
     6421 ggaggcagga gggcgtggga gctctggaga ttaaggtccc cttcgccatc accctgcaca
     6481 tcaagggcag caacgagagc gtagtgatcc ggcagaaaag catcaagctg aaaaactacg
     6541 gcgacttcca caagtttctg tacgataacc ggatcgagac cctgatccct tacctgcggg
     6601 acgacgccaa tgccgtgaag gtgaccattg atagggatga actggaacac gagtttgaga
     6661 cctatgataa ggtgaggctg gacgtgtttg aatctgtgca caagatcgag aggctgattc
     6721 tggagcggta ccccgagctg cacgacaaaa ggagctccct gtactatttt gagaagaagg
     6781 gcgtggagaa gccaatcccc aagaagaata atttcaggca gctgatcgag tgcgagcccc
     6841 agttcgccgt gctggacaag gatgactgcc tgaacatcgc caacagcttc agcgccaact
     6901 cctacaattc taaggaccat gagaaggtga acatcccaag cggcaaaatc ggcgagatcg
     6961 ccgagctgat ctccgagcgg atgaaaacct gcctgaacaa cttcgaaaag aaaacccgcc
     7021 aggagaatga ctccgagagt ctagaggcgc taccggtcgc caccatggtg agcaagggcg
     7081 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     7141 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     7201 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     7261 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     7321 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     7381 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     7441 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     7501 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     7561 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     7621 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     7681 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     7741 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtcgaattct aagccaggat
     7801 tccatatgaa gcgacctgcc gccacaaaga aggctggaca ggctaagaag aagaaacaac
     7861 gcgtgcagct aagcgactac aaagaccatg acggtgatta taaagatcat gacatcgatt
     7921 acaaggatga cgatgacaag taaggatccc tccccccccc ctaacgttac tggccgaagc
     7981 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     8041 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     8101 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     8161 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     8221 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     8281 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     8341 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     8401 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     8461 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataaggatcc
     8521 accggagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     8581 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     8641 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     8701 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     8761 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     8821 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     8881 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     8941 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     9001 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     9061 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     9121 gcccggtgcc tgacgcccat cgatagatcc taatcaacct ctggattaca aaatttgtga
     9181 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     9241 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     9301 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     9361 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     9421 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     9481 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     9541 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     9601 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     9661 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     9721 cctttgggcc gcctccccgc ctgagatcct ttaagaccaa tgacttacaa ggcagctgta
     9781 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     9841 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     9901 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     9961 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    10021 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
    10081 ttataacttg caaagaaatg aatatcagag agtgagaggc ccgggttaat taaggaaagg
    10141 gctagatcat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
    10201 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
    10261 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
    10321 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
    10381 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
    10441 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
    10501 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
    10561 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
    10621 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
    10681 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
    10741 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
    10801 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
    10861 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
    10921 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
    10981 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
    11041 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
    11101 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
    11161 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
    11221 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
    11281 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    11341 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
    11401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    11461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    11521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    11581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    11641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    11701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    11761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    11821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    11881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    11941 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
    12001 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
    12061 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
    12121 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
    12181 tctccccgcg cgttggccga ttcattaatg cagcaagctc atggctgact aatttttttt
    12241 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
    12301 ttttttggag gcctaggctt ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg
    12361 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
    12421 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
    12481 tcacacagga aacagctatg acatgattac gaatttcaca aataaagcat ttttttcact
    12541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
    12601 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
    12661 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
    12721 aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
