LOCUS       sequence-478684-       13137 bp    DNA     circular SYN 15-JUL-2025
DEFINITION  Overexpression of dAsCas12a  in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13137)
  AUTHORS   Yang LZ, Min YH, Liu YX, Gao BQ, Liu XQ, Huang Y, Wang H, Yang L, 
            Liu ZJ, Chen LL
  TITLE     CRISPR-array-mediated imaging of non-repetitive and multiplex 
            genomic loci in living cells.
  JOURNAL   Nat Methods. 2024 Sep;21(9):1646-1657. doi: 
            10.1038/s41592-024-02333-3. Epub 2024 Jul 4.
  PUBMED    38965442
REFERENCE   2  (bases 1 to 13137)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02333-3"; journalName: "Nat Methods"; date: 
            "2024-09"; volume: "21"; issue: "9"; pages: "1646-1657"
COMMENT     Sequence Label: pHAGE-IRES-puro-NLS-dAsCas12a-EGFP-NLS-3xFlag
FEATURES             Location/Qualifiers
     source          1..13137
                     /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             7392..7412
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7434..7443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7440..7442,7443..7445,7446..8156)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7440..7442,7443..7445,7446..8156)
                     /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(7485..7506)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7746..7765)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8093..8114
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8184..8231
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8250..8315
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    8322..8889
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8483..8500)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8710..8729
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8909..9508
                     /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(8909..8928)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9405..9425
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9527..10115
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9580..9600)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9998..10009)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10190..10423
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10542..10560)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10628..10732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10733..10801,10802..11593)
                     /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(10951..10970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11764..12352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12253..12272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12506..12523
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12763..12784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12799..12804,12805..12822,12823..12829)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12837..12853
                     /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     12842..12864
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12861..12877
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12861..12877
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12927..12946
                     /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 agctagcact cagttcgagg
     3481 gcttcacaaa cctgtatcaa gtgtccaaga ccctccgctt cgagctgatc cctcagggta
     3541 agaccctgaa gcatatacag gagcagggtt tcatcgagga ggacaaggcc aggaatgatc
     3601 actacaagga gctgaagccc atcattgatc gcatctacaa gacatacgca gatcaatgtc
     3661 tgcaactcgt ccagctggat tgggagaatc tgtcagccgc aatcgacagt tatcggaaag
     3721 agaagacaga agaaaccagg aacgccctca tcgaggaaca agcaacctac cggaatgcaa
     3781 tacacgacta tttcatcggc aggactgaca atctgactga cgccatcaat aagagacacg
     3841 cagaaatcta taagggcctc tttaaggccg aactcttcaa cggcaaagtc ctcaaacagc
     3901 tgggcacagt taccactacc gagcacgaga atgccctgct ccggagcttc gacaagttca
     3961 ccacttactt ctctgggttc tacgagaatc ggaagaacgt gttcagcgca gaggacatct
     4021 ccacagccat tccacaccgg atcgtccagg ataactttcc caaattcaaa gagaattgcc
     4081 atatctttac caggctgatc actgccgttc catctctgag ggaacacttt gagaacgtga
     4141 agaaagccat cgggatattc gtgtctactt ccatcgagga agtgttctct ttccctttct
     4201 acaatcagct gctgacccag acccagatcg acctctataa ccagctgctg ggaggtatca
     4261 gcagggaggc aggcaccgag aagatcaagg gactgaatga ggtcctgaac ctcgccattc
     4321 agaagaacga tgagacagca cacatcatcg cctctctgcc acatcgcttc attcctctgt
     4381 tcaaacagat tctctctgat cggaacaccc tcagctttat cctggaggag tttaagtccg
     4441 atgaagaggt tattcagtct ttctgtaagt acaagactct gctgcgcaat gagaacgtgc
     4501 tggagactgc cgaagctctg tttaacgaac tgaactccat cgacctcacc cacatcttca
     4561 tcagccacaa gaagctggag acaattagca gtgccctgtg tgaccattgg gataccctgc
     4621 gcaacgccct gtacgagagg agaatatccg agctgacagg taagataacc aagagcgcca
     4681 aagagaaagt ccaaaggtcc ctgaaacatg aggacataaa cctgcaggag attatctcag
     4741 cagctggtaa ggaactctca gaggctttca agcagaagac ctccgaaatt ctctcccatg
     4801 cccacgcagc actcgatcaa ccactgccaa ccaccctgaa gaaacaagag gagaaggaaa
     4861 tcctgaagag ccagctggac agcctcctcg ggctgtacca tctgctcgat tggttcgccg
     4921 tggacgaaag taacgaggtg gacccagagt tctcagcaag actgaccggc atcaagctgg
     4981 agatggagcc cagtctctcc ttctacaaca aagctagaaa ctacgccacc aagaagcctt
     5041 attctgtgga gaagtttaag ctgaacttcc agatgcccac tctggcttcc ggttgggacg
     5101 tcaacaagga gaagaataac ggtgcaatcc tgttcgtgaa gaacggactg tactacctgg
     5161 gcatcatgcc aaagcagaag ggtcgctaca aagccctctc atttgaaccc acagagaaga
     5221 caagcgaagg atttgacaag atgtattacg attactttcc cgatgccgct aagatgatcc
     5281 ctaagtgcag cacccagctg aaggccgtca ccgctcattt ccagacccat actacaccaa
     5341 ttctgctgtc taacaacttc atagagccac tggagataac caaggaaatc tacgatctga
     5401 acaatccaga gaaggagccc aagaagtttc agaccgccta cgctaagaag accggagatc
     5461 agaaaggtta tcgggaggcc ctgtgcaaat ggatcgactt cacacgcgat ttcctcagca
     5521 agtatacaaa gaccactagc atagacctgt ccagcctcag gccctcaagt cagtacaagg
     5581 atctcggtga gtactacgcc gaactcaacc cactgctcta ccacattagc tttcagagga
     5641 tcgccgagaa agagatcatg gatgcagtgg agactggcaa gctgtatctg tttcagatat
     5701 acaacaaaga cttcgctaaa ggacaccacg gtaagccaaa cctgcatact ctctattgga
     5761 ccggactctt cagtccagag aatctcgcca agacctcaat caagctgaac ggtcaggcag
     5821 agctgttcta tagacctaag agtaggatga aacggatggc acatagactc ggcgagaaga
     5881 tgctgaacaa gaagctgaaa gaccagaaga ctcccattcc agatacactg taccaggagc
     5941 tgtatgatta cgtgaatcac cggctctccc acgacctgtc tgatgaagcc agggctctcc
     6001 tgcctaacgt gataactaaa gaggtgagcc atgagatcat caaagaccgg agattcacat
     6061 ccgacaagtt cttctttcat gttccaatta cactgaacta tcaagcagca aattctccca
     6121 gcaagtttaa ccaaagggtg aacgcctacc tgaaggaaca tcccgaaaca cccatcattg
     6181 gaattgctag aggtgagcgc aatctcatct acataaccgt gatcgactcc acaggtaaga
     6241 ttctcgagca gaggtccctc aacactattc agcagtttga ttatcagaag aaactcgaca
     6301 acagagagaa agagagagtc gctgccagac aagcatggag tgtcgttggc acaatcaaag
     6361 acctgaagca gggatacctg agccaagtta tacacgaaat tgtggatctg atgatccact
     6421 accaggctgt tgtggtgctg gcgaacctca actttggttt caagtctaag aggacaggga
     6481 tcgctgagaa agctgtgtac cagcagtttg agaagatgct cattgataaa ctgaactgtc
     6541 tcgtgctcaa agactaccct gccgagaagg tgggaggagt cctgaatcct tatcagctga
     6601 ctgaccagtt tacctccttc gctaagatgg gcacccaatc aggctttctg ttctacgtgc
     6661 ctgcacccta caccagtaag attgatccac tgacagggtt cgttgatcct ttcgtgtgga
     6721 agaccataaa gaaccacgaa agcagaaagc acttcctgga aggcttcgac ttcctgcatt
     6781 atgacgtcaa gactggagac ttcatactgc atttcaagat gaatagaaac ctgtcatttc
     6841 agagaggact gcctgggttc atgccagctt gggacattgt ctttgagaag aatgagacac
     6901 agtttgacgc caaaggcact ccattcattg ctggcaagcg gatcgtccct gtgatagaga
     6961 accacagatt cactggaaga tacagggacc tgtacccagc aaatgaactc attgccctcc
     7021 tggaagagaa agggatcgtg ttccgggatg ggtctaacat cctgccaaag ctcctggaga
     7081 atgacgattc acatgccatt gatacaatgg ttgccctgat ccgcagtgtg ctgcaaatga
     7141 ggaatagcaa tgcagccacc ggagaggcct acatcaatag tccagttcgg gacctcaatg
     7201 gcgtgtgttt cgatagtaga ttccagaacc ctgagtggcc tatggacgct gatgctaacg
     7261 gtgcttatca tattgccctg aagggccagc tcctgctgaa ccacctcaag gagtctaagg
     7321 acctgaagct ccagaatggc atttctaatc aggattggct ggcctatatc caggaactga
     7381 ggaacggcag tcccaagaag aagaggaagg tgagtctaga ggcgctaccg gtcgccacca
     7441 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     7501 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     7561 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     7621 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     7681 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     7741 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     7801 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     7861 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     7921 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     7981 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     8041 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     8101 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtcga
     8161 attctaagcc aggattccat atgaagcgac ctgccgccac aaagaaggct ggacaggcta
     8221 agaagaagaa acaacgcgtg cagctaagcg actacaaaga ccatgacggt gattataaag
     8281 atcatgacat cgattacaag gatgacgatg acaagtaagg atccctcccc cccccctaac
     8341 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     8401 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     8461 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     8521 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     8581 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     8641 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     8701 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     8761 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     8821 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     8881 cgatgataag gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     8941 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     9001 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     9061 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     9121 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     9181 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     9241 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     9301 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     9361 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     9421 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     9481 gtgcatgacc cgcaagcccg gtgcctgacg cccatcgata gatcctaatc aacctctgga
     9541 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     9601 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     9661 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     9721 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     9781 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     9841 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     9901 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     9961 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
    10021 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
    10081 gacgagtcgg atctcccttt gggccgcctc cccgcctgag atcctttaag accaatgact
    10141 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
    10201 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
    10261 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
    10321 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
    10381 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
    10441 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggcccggg
    10501 ttaattaagg aaagggctag atcattcttg aagacgaaag ggcctcgtga tacgcctatt
    10561 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
    10621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
    10681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
    10741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
    10801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
    10861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
    10921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
    10981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
    11041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
    11101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
    11161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
    11221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
    11281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
    11341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
    11401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
    11461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
    11521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
    11581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
    11641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
    11701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    11761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
    11821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
    11881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
    11941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
    12001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
    12061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
    12121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
    12181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    12241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
    12301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
    12361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
    12421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
    12481 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
    12541 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagca agctcatggc
    12601 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
    12661 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc cgtggcacga
    12721 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
    12781 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
    12841 gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt tcacaaataa
    12901 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
    12961 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
    13021 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
    13081 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
