LOCUS       sequence-375392-       12975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of NLS-dMisCas13b-EGFP-NLS in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12975)
  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 12975)
  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-IRES-puro-NLS-dMisCas13b-EGFP-NLS
FEATURES             Location/Qualifiers
     source          1..12975
                     /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      7272..7281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7278..7280,7281..7283,7284..7994)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7278..7280,7281..7283,7284..7994)
                     /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(7584..7603)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7931..7952
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8022..8069
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8088..8153
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    8160..8727
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8321..8338)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8548..8567
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8747..9346
                     /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(8747..8766)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9243..9263
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9365..9953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9418..9438)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9836..9847)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10028..10261
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10380..10398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10466..10570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10571..10639,10640..11431)
                     /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(10789..10808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11602..12190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12091..12110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12344..12361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12601..12622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12637..12642,12643..12660,12661..12667)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12675..12691
                     /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     12680..12702
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12699..12715
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12699..12715
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12765..12784
                     /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 agtctagagg
     7261 cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     7321 tcctggtcga gctggacgga gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     7381 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     7441 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     7501 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     7561 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     7621 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     7681 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     7741 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     7801 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     7861 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     7921 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     7981 acgagctgta caagtcgaat tctaagccag gattccatat gaagcgacct gccgccacaa
     8041 agaaggctgg acaggctaag aagaagaaac aacgcgtgca gctaagcgac tacaaagacc
     8101 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagtaaggat
     8161 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     8221 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     8281 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     8341 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     8401 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     8461 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     8521 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     8581 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     8641 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     8701 gttttccttt gaaaaacacg atgataagga tccaccggag cttaccatga ccgagtacaa
     8761 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     8821 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     8881 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     8941 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     9001 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     9061 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     9121 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     9181 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     9241 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     9301 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc catcgataga
     9361 tcctaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     9421 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     9481 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     9541 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     9601 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     9661 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     9721 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     9781 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     9841 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     9901 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat
     9961 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
    10021 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
    10081 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
    10141 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
    10201 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
    10261 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
    10321 gagagtgaga ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg
    10381 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
    10441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
    10501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
    10561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
    10621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
    10681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
    10741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
    10801 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
    10861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
    10921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
    10981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
    11041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
    11101 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
    11161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
    11221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
    11281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
    11341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
    11401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
    11461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
    11521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
    11581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
    11641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
    11701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
    11761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
    11821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
    11881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
    11941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
    12001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
    12061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
    12121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
    12181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
    12241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
    12301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
    12361 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
    12421 atgcagcaag ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
    12481 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
    12541 aagctccccg tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
    12601 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
    12661 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
    12721 tacgaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    12781 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
    12841 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta
    12901 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac
    12961 tacaaactta gtagt
//
