LOCUS       sequence-494131-       12825 bp    DNA     circular SYN 01-DEC-2025
DEFINITION  Overexpression of HyperdLbCas12a in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12825)
  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 12825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 1, 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-HyperdLbCas12a-tagBFP-NLS-...
FEATURES             Location/Qualifiers
     source          1..12825
                     /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             7158..7865
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDM
                     ALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     CDS             7872..7919
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7938..8003
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    8010..8577
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8171..8188)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8398..8417
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8597..9196
                     /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(8597..8616)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9093..9113
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9215..9803
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9268..9288)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9686..9697)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9878..10111
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10230..10248)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10316..10420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10421..10489,10490..11281)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10639..10658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11452..12040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11941..11960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12194..12211
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12451..12472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12487..12492,12493..12510,12511..12517)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12525..12541
                     /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     12530..12552
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12549..12565
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12549..12565
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12615..12634
                     /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 agctagctca aagctggaga
     3481 aattcaccaa ctgttatagc ctgtctaaga ccctgcgctt caaggcaatc ccagtgggca
     3541 agacacaaga gaacattgac aacaaacggc tcctggtgga ggatgagaag agggctgaag
     3601 attacaaggg cgttaagaag ctgctggata ggtactatct gtcattcatc aacgatgtcc
     3661 tccacagtat caagctgaag aatctgaaca attacatttc tctgttccgg aagaagacac
     3721 ggaccgagaa ggagaacaaa gagctggaga atctggagat caacctgagg aaagaaatag
     3781 ctaaggcttt caaagggaac gagggttaca agtccctgtt caagaaagac attatcgaga
     3841 ctattctgcc tgagttcctg gacgataaag atgagatcgc cctcgtcaat tccttcaatg
     3901 ggtttaccac agcctttacc ggcttcttca ggaatagaga gaatatgttc tctgaagagg
     3961 ccaaatccac tagcatcgcc tttcgctgca taaacgagaa cctgactagg tacatcagca
     4021 atatggacat ctttgagaaa gtcgatgcca tattcgacaa acatgaggtg caggagatta
     4081 aggagaagat cctgaactca gattacgatg tcgaagattt cttcgaggga gagttcttca
     4141 acttcgtgct cacacaagag ggcattaggg tgtacaatgc aatcattgga gggttcgtga
     4201 cagagagtgg cgagaagata aagggcctga acgagtatat caacctctac aaccagaaaa
     4261 ccaagcagaa actgcctaag ttcaagccac tgtacaaaca agtgctctca gatagggaaa
     4321 gcctgagctt ctacggtagg gggtatacat cagatgaaga agtgctcgaa gtgttccgca
     4381 acaccctcaa taagaacagt gaaatcttct cttcaatcaa gaagctggag aaactgttca
     4441 agaatttcga tgagtactcc tctgccggaa tctttgtgaa gaatggccct gcaatatcca
     4501 ctattagcaa aaggatcttt ggcgagtgga acgttatcag ggataagtgg aatgccgagt
     4561 acgatgatat tcatctcaag aagaaagccg tggttacaga gaaatacgag gatgatagac
     4621 gcaagagctt taagaagatt ggtagcttct ctctcgaaca gctgcaggag tacgccgacg
     4681 ctgacctgtc agtcgtggag aaactcaagg agatcataat ccagaaggtg gatgaaatct
     4741 acaaagtgta tggaagctct gagaaactct tcgatgcaga ctttgttctg gagaagagtc
     4801 tgaagaagaa cgacgcagtg gttgctatca tgaaggacct gctggattct gttaagtctt
     4861 tcgagaatta cattaaggca ttctttggtg aagggaagga gacaaatagg gacgagagct
     4921 tctatggcga ctttgttctg gcctacgaca tcctcctcaa ggttgaccac atctatgacg
     4981 ctatacggaa ttacgttacc cagaagccct atagcaaaga caagttcaag ctgtatttcc
     5041 agaatccaca gtttatgggt gggtgggata aagacaaaga aacagattac agggccacta
     5101 tcctgcggta cggcagcaaa tactatctgg ctatcatgga taagaagtac gccaaatgcc
     5161 tccagaagat cgacaaggac gacgtgaacg gtaactacga gaagatcaat tacaagctcc
     5221 tgccaggacc taacaagatg ctgcccaagg tgttcttctc caagaaatgg atggcctact
     5281 ataacccaag cgaggacatt cagaagatat acaagaatgg gacattcaag aagggcgata
     5341 tgttcaacct caacgactgc cacaagctga ttgatttctt caaggatagc atttctcgct
     5401 atcccaagtg gtctaatgca tacgatttca acttcagcga gactgagaag tacaaagaca
     5461 tcgctggctt ctaccgggag gtggaagagc aaggctataa ggtgtcattc gaatccgctt
     5521 ctaagaagga agtggataag ctcgtggaag agggtaagct gtacatgttc cagatataca
     5581 acaaagactt cagcgataag agccacggca ctccaaacct ccatactatg tatttcaagc
     5641 tgctgtttga cgagaacaac cacggacaga ttaggctgtc aggaggcgca gaactcttca
     5701 tgcgcagagc ttcactgaag aaggaggaac tcgttgtcca cccagccaat agccctatag
     5761 ccaataagaa tccagacaat cctaagaaaa ccactactct gtcttacgat gtgtataagg
     5821 ataagagatt ctctgaagat cagtacgaac tgcacatacc cattgccatt aacaagtgcc
     5881 ctaagaacat cttcaagatt aacacagagg ttagagtgct cctgaaacac gacgataacc
     5941 cttatgttat aggcattgct cgcggagaga gaaacctgct gtacatcgtc gtggtggacg
     6001 gcaaaggcaa catcgtggaa cagtacagtc tcaatgaaat cattaacaat ttcaacggaa
     6061 tccgcattaa gaccgactac cattctctcc tcgacaagaa ggagaaagaa aggttcgaag
     6121 caagacagaa ttggacaagt atagagaata tcaaagaact gaaggctggg tacatctctc
     6181 aggttgtgca caagatatgt gagctggtgg agaagtacga cgctgttatc gccctcgcgg
     6241 acctgaatag cggcttcaag aactccaggg tgaaggtgga gaagcaggtg tatcagaagt
     6301 tcgagaagat gctgatcgac aagctcaact atatggtgga caagaaatcc aatccttgcg
     6361 ctactggtgg agccctgaag ggctatcaaa tcaccaataa gttcgaatct ttcaagtcta
     6421 tgagcaccca gaatggcttc atcttctaca tacccgcatg gctgacatcc aagattgatc
     6481 cctctaccgg atttgttaat ctgctcaaga ctaagtacac ctctattgct gactcaaaga
     6541 agttcatatc atcatttgac cgcatcatgt acgtgccaga agaggacctg ttcgagtttg
     6601 ccctggatta caagaatttc tctcggactg acgccgacta catcaagaag tggaagctct
     6661 actcttatgg taatcggatt cgcatattcc gcaatcccaa gaagaataac gtgttcgatt
     6721 gggaggaagt ttgcctcacc agcgcttaca aggagctgtt caataagtat gggattaact
     6781 accagcaggg cgacataaga gccctgctgt gcgaacaatc tgataaggca ttctattcct
     6841 ctttcatggc actgatgtca ctgatgctgc aaatgcgcaa ttccatcacc ggaagaacag
     6901 acgtggcctt tctgatctct cctgtcaaga actcagatgg catcttctac gattcccgca
     6961 actatgaagc acaggagaat gctatcctgc ctaagaatgc cgatgcaaat ggagcctata
     7021 acatcgccag aaaggtcctc tgggccatag gacaattcaa gaaagctgaa gatgagaagc
     7081 tggacaaggt gaagatcgcc atttcaaaca aagagtggct cgaatatgct cagacctcag
     7141 tgaagcatag tctagaggtg tctaagggcg aagagctgat taaggagaac atgcacatga
     7201 agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag
     7261 gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc
     7321 ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc
     7381 acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga
     7441 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
     7501 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg
     7561 tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg
     7621 gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg
     7681 caaacgccaa gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg
     7741 tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgaa acctacgtcg
     7801 agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc
     7861 ttaatcatat gaagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaac
     7921 aacgcgtgca gctaagcgac tacaaagacc atgacggtga ttataaagat catgacatcg
     7981 attacaagga tgacgatgac aagtaaggat ccctcccccc cccctaacgt tactggccga
     8041 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     8101 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     8161 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     8221 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     8281 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     8341 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     8401 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     8461 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     8521 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagga
     8581 tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     8641 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     8701 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     8761 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     8821 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     8881 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     8941 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     9001 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     9061 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     9121 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     9181 caagcccggt gcctgacgcc catcgataga tcctaatcaa cctctggatt acaaaatttg
     9241 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     9301 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9361 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9421 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9481 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     9541 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     9601 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     9661 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     9721 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     9781 ctccctttgg gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct
     9841 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     9901 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     9961 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
    10021 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
    10081 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
    10141 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa
    10201 agggctagat cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
    10261 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
    10321 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
    10381 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
    10441 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
    10501 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
    10561 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
    10621 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
    10681 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
    10741 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
    10801 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
    10861 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
    10921 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
    10981 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
    11041 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
    11101 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
    11161 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
    11221 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
    11281 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
    11341 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
    11401 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    11461 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    11521 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    11581 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    11641 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    11701 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    11761 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    11821 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    11881 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    11941 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    12001 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    12061 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    12121 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    12181 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
    12241 gcctctcccc gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt
    12301 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
    12361 ggcttttttg gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga
    12421 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
    12481 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
    12541 atttcacaca ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc
    12601 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
    12661 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
    12721 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt
    12781 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
