LOCUS       sequence-494130-       12855 bp    DNA     circular SYN 01-DEC-2025
DEFINITION  Overexpression of dLbCas12a in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12855)
  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 12855)
  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-dLbCas12a-EGFP-NLS-3xFlag
FEATURES             Location/Qualifiers
     source          1..12855
                     /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             join(7158..7160,7161..7163,7164..7874)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7158..7160,7161..7163,7164..7874)
                     /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(7203..7224)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7464..7483)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7811..7832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7902..7949
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7968..8033
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    8040..8607
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8201..8218)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8428..8447
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8627..9226
                     /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(8627..8646)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9123..9143
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9245..9833
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9298..9318)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9716..9727)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9908..10141
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10260..10278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10346..10450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10451..10519,10520..11311)
                     /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(10669..10688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11482..12070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11971..11990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12224..12241
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12481..12502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12517..12522,12523..12540,12541..12547)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12555..12571
                     /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     12560..12582
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12579..12595
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12579..12595
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12645..12664
                     /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 ggcttcttcg acaatagaga 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 ggcattgatg tgtacaatgc aatcattgga gggttcgtga
     4201 cagagagtgg cgagaagata aagggcctga acgagtatat caacctctac aaccagaaaa
     4261 ccaagcagaa actgcctaag ttcaagccac tgtacaaaca agtgctctca gatagggaaa
     4321 gcctgagctt ctacggtgaa gggtatacat cagatgaaga agtgctcgaa gtgttccgca
     4381 acaccctcaa taagaacagt gaaatcttct cttcaatcaa gaagctggag aaactgttca
     4441 agaatttcga tgagtactcc tctgccggaa tctttgtgaa gaatggccct gcaatatcca
     4501 ctattagcaa agacatcttt 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 tgaagcatgg atcacccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     7201 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     7261 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     7321 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     7381 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     7441 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     7501 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     7561 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     7621 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     7681 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     7741 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     7801 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     7861 acgagctgta caagtcgaat tctaagccag gattccatat gaagcgacct gccgccacaa
     7921 agaaggctgg acaggctaag aagaagaaac aacgcgtgca gctaagcgac tacaaagacc
     7981 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagtaaggat
     8041 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     8101 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     8161 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     8221 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     8281 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     8341 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     8401 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     8461 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     8521 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     8581 gttttccttt gaaaaacacg atgataagga tccaccggag cttaccatga ccgagtacaa
     8641 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     8701 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     8761 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     8821 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     8881 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     8941 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     9001 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     9061 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     9121 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     9181 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc catcgataga
     9241 tcctaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     9301 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     9361 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     9421 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     9481 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     9541 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     9601 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     9661 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     9721 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     9781 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat
     9841 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     9901 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     9961 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
    10021 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
    10081 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
    10141 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
    10201 gagagtgaga ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg
    10261 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
    10321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
    10381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
    10441 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
    10501 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
    10561 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
    10621 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
    10681 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
    10741 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
    10801 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
    10861 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
    10921 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
    10981 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
    11041 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
    11101 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
    11161 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
    11221 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
    11281 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
    11341 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
    11401 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
    11461 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
    11521 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
    11581 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
    11641 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
    11701 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
    11761 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
    11821 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
    11881 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
    11941 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
    12001 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
    12061 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
    12121 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
    12181 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
    12241 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
    12301 atgcagcaag ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
    12361 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
    12421 aagctccccg tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
    12481 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
    12541 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
    12601 tacgaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    12661 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
    12721 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta
    12781 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac
    12841 tacaaactta gtagt
//
