LOCUS       sequence-375412-       12765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of NLS-dMisCas13b-SNAPf-NLS-3xFlag in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12765)
  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 12765)
  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"; 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-NLS-dMisCas13b-SNAPf-NLS-3xFlag
FEATURES             Location/Qualifiers
     source          1..12765
                     /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             7260..7805
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             7812..7859
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7878..7943
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    7950..8517
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8111..8128)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8338..8357
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8537..9136
                     /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(8537..8556)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9033..9053
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9155..9743
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9208..9228)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9626..9637)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9818..10051
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10170..10188)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10256..10360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10361..10429,10430..11221)
                     /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(10579..10598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11392..11980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11881..11900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12134..12151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12391..12412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12427..12432,12433..12450,12451..12457)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12465..12481
                     /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     12470..12492
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12489..12505
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12489..12505
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12555..12574
                     /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 ttcgtgagat
     3481 atagccctga gtcctggcct aggcacttcc acggcctgca gcagagactg cccaccccca
     3541 tcggccagca caccttcctg agtgataact ttaacaccat ggacatccgg ctgagtaaga
     3601 atagagccct gctggcagct tacctgaata tggcccagca gaacgtgtac gtgaccctgc
     3661 tgcacatctc ccagctgatg ggagtggagg agaccaaaga cctgtctaac cgcatgtacg
     3721 actttcagat tctgcaggag cttaagaccg ggaagaaggg cgccgagtct gagcagatgg
     3781 tgaagtctga gagagtgcag aagatcattc tcctgctgta tcgcaacttc cctttcctga
     3841 aggccttcgt ggaggctaag tacaagcagc aggagaagaa caatgggggc aaaaacaagg
     3901 gcggcggaaa aaacacacag aagaataaga aaacccctga caatacaagc aacgaggagc
     3961 agctggagtt cactgagagt gagctgggca agctgcccat tccaccacat gagtacagaa
     4021 ccatcctgtg tgatgtgttc aggctgatca atgacctggc caatgagtac acagccttca
     4081 gccctgagaa acgcactggc ggcgctaagg agtcccacga aaagtgccag aagaagatgc
     4141 tgagctacct gtacctgtgc ctggacggag gcagaagggt ggtgaaggag cggttcaagg
     4201 acaccttcca agagaaggac tttaactttc tcacccaggg cccaagatac agcctccaga
     4261 aagacggaga taagaagaag gccgaggaaa gacctaattt ttactacaga ctgaaggatg
     4321 accagggact gttctcttac gtcggtatcc tcctgttcat cagcctgttc ctgcacaaga
     4381 ggtatatttc cgagatgctg gcccagtccg ggctgtttta tacacctcgg tactggaagg
     4441 gccagccagt gaacgtggaa ctggagagga ggatcatcag tgagatcttt tccatctatc
     4501 gcatcaggct gcccaaggag agaatcgact ccaccagacc agattacgcc ctcggcctgg
     4561 atatgctgaa tgagctgcag aagtgcccta atgagctgta tgagaccttc tcacctaaag
     4621 accaggctga gtttagagtg aagcgggagt atgacgagag cggcgaaggc acagaagagg
     4681 gcgaggtgct gctgaaaaga ttcgaggaca gattccctta tttcgccatt cgctatctgg
     4741 acgagaatag agtgttcgac gacatcagat tccaggtgag catcggcaag tataggtaca
     4801 aattctacaa taagctgtgc atcgacgtgg tgagcagcaa tcggaacgat gactgctatc
     4861 gcctgaggtc cctgcagaaa gagattaatg gcttcggcag actgaatgaa gtggagtacg
     4921 agagaatcct gagatacggg tctcacatcc acgtggagaa gcctgagggc gacgacaccc
     4981 cctacctgac tgaccatcat accagctacc tgtttaacgg caataggatc ggcctgctgt
     5041 tcaaccagga gaacggcagg cacgatctgg aggagatcag aggaaactgc ctgtatctgc
     5101 ctcacctgac ttttgcatcc tcctccgagg gaaaggatgc caggcgcgtg ccccccatcg
     5161 cctggctgtc cacctatgag ctgcctgccc tgatcttcca ccacctgctg tgcagacact
     5221 atggcctgga gagaactgcc accgagagca tcatcaagga atgtgtgggc ctgtaccacc
     5281 ggctgtttac tgatatccgc gacggtaaac tgctccccgg agaggccctg ggcatccagg
     5341 ccgaggagtc cacaaggagc gagattagta ggatcaagaa gggtgaaccc ctgccccaga
     5401 acatctctga aaagattgaa accttcctga gcactcacta ctgcaccgaa caggtgtgcc
     5461 tgcacatcca ggacattcct aagaaaatca tcgattacct gatcgggaaa aacatcgacc
     5521 aggacatgag cggactcttt aagcagagaa gcagagagcg gattaataag atgtatgagt
     5581 ccactaaaat tcgcctcaag aagttcaaga ccgacctgga cctgcagggc aacggcgaca
     5641 ataagtttgg aaagaagagg tacactgaca ttaggccagg acagctggcc gactatctgg
     5701 ccaaggacct gctgttcttc cagccatatg acgacgagaa gaagaataag gtcactggac
     5761 tgaactatca gatcctgcag gccgcactgg ccatctacaa tcacaacctg gatgagctca
     5821 gacgactctt caaaaatgcc cggctgctcg tgcccgagaa tcctaagatg gagcaccctt
     5881 tcctgggctc tgtgctgcac aagtcccccc ggaccactct gcagttctac gcctactacc
     5941 tgctcgagaa gatctcttac ctgaaaaaga gactcgacga tcctaacgag acattcgccg
     6001 atctgtttag atataagcag aggcagaaat ggcagactag agacagccag tggtataggg
     6061 ccttggctgg caggtacatg gctgagatga tggaagaggg caccatgaat tttaaacccg
     6121 tggagctgcc aagagggctg tttgaaagac ctatcaagcg gctgctgacc aatatcctga
     6181 ctaagcggac cgacagcgat agccaggccc tgctgaagat cgtggagagc cccaggtgca
     6241 acgtgagcta cctgattgct cagtatttca ctctgattct gcacgacgat tcccagccct
     6301 tttaccaccc taacaaacgc tacaaaagga cctaccagta tttcaacatc ctgaacaata
     6361 gaaaggagcg gaataagctc gtgcaggtgt actacactct ggatgagatg gaggcttact
     6421 ttaaggatac cgaatctctg aagaagaggt ataaggacta cctgactagc tccgaggcta
     6481 agaagcaccc agcccccgag cagaggctga agaagtatct gtcagagtat cgcaagaatg
     6541 agaccaccat tagacggtac agagtgcagg acatgctggt gtttctgatg gccaaaattc
     6601 tgctgctgga aaagggcggc ctcgaagccg gcgagctgga gcagtacaag ctgtcccaca
     6661 ttcagcccac caccggccag aacagcggcc agaacatcct gagcctgcag gtgcccttta
     6721 acatcaccgt gaagctggac gatggcacag agagaagcat cacccaggaa gtgatcaagg
     6781 tgaaaaatta tggcgacttc tttagatttc tctatgatga ccgggtggca tccctgctgc
     6841 cttatactca ggaggacatt cccctggagc gggccctgct ggagaaggag ctggagggat
     6901 acgacaggga gagaatccct attttcaaga gcgtgcactg catggagaag gagatcctga
     6961 ggctgtgccc tgaactgaat gatccccagt ccagccagta caagtatacc acagatattg
     7021 gcactgagaa gatcgtgaag gagaatttca gggccatgct gactgagctg tgtagccggg
     7081 gcctgctccc tgagaacgag atcgagatga tggtggagat cgccaatagc ttttccgcca
     7141 atagatacgc ctgcgtgaac ctgaagggca agaatatccc aatgatcgcc atctccatcc
     7201 agcggaagtt cgacggcgtg gtgggcagaa ccatcggaaa gaataagaac agtctagaaa
     7261 tggacaaaga ctgcgaaatg aagcgcacca ccctggatag ccctctgggc aagctggaac
     7321 tgtctgggtg cgaacagggc ctgcaccgta tcatcttcct gggcaaagga acatctgccg
     7381 ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg cggaccagag ccactgatgc
     7441 aggccaccgc ctggctcaac gcctactttc accagcctga ggccatcgag gagttccctg
     7501 tgccagccct gcaccaccca gtgttccagc aggagagctt tacccgccag gtgctgtgga
     7561 aactgctgaa agtggtgaag ttcggagagg tcatcagcta cagccacctg gccgccctgg
     7621 ccggcaatcc cgccgccacc gccgccgtga aaaccgccct gagcggaaat cccgtgccca
     7681 ttctgatccc ctgccaccgg gtggtgcagg gcgacctgga cgtggggggc tacgagggcg
     7741 ggctcgccgt gaaagagtgg ctgctggccc acgagggcca cagactgggc aagcctgggc
     7801 tgggtcatat gaagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaac
     7861 aacgcgtgca gctaagcgac tacaaagacc atgacggtga ttataaagat catgacatcg
     7921 attacaagga tgacgatgac aagtaaggat ccctcccccc cccctaacgt tactggccga
     7981 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     8041 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     8101 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     8161 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     8221 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     8281 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     8341 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     8401 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     8461 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagga
     8521 tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     8581 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     8641 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     8701 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     8761 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     8821 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     8881 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     8941 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     9001 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     9061 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     9121 caagcccggt gcctgacgcc catcgataga tcctaatcaa cctctggatt acaaaatttg
     9181 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     9241 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9301 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9361 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9421 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     9481 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     9541 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     9601 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     9661 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     9721 ctccctttgg gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct
     9781 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     9841 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     9901 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     9961 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
    10021 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
    10081 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa
    10141 agggctagat cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
    10201 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
    10261 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
    10321 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
    10381 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
    10441 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
    10501 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
    10561 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
    10621 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
    10681 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
    10741 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
    10801 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
    10861 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
    10921 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
    10981 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
    11041 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
    11101 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
    11161 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
    11221 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
    11281 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
    11341 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    11401 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    11461 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    11521 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    11581 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    11641 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    11701 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    11761 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    11821 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    11881 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    11941 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    12001 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    12061 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    12121 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
    12181 gcctctcccc gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt
    12241 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
    12301 ggcttttttg gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga
    12361 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
    12421 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
    12481 atttcacaca ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc
    12541 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
    12601 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
    12661 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt
    12721 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
