LOCUS       sequence-175378-       12104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Nm dCas9-3XmCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12104)
  AUTHORS   Ma H, Naseri A, Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T
  TITLE     Multicolor CRISPR labeling of chromosomal loci in human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7. doi: 
            10.1073/pnas.1420024112. Epub 2015 Feb 23.
  PUBMED    25713381
REFERENCE   2  (bases 1 to 12104)
  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"; doi: 
            "10.1073/pnas.1420024112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-10- 10"; volume: "112"; issue: "10"; pages: "3002-7"
COMMENT     Sequence Label: pHAGE-TO-nmdCas9-3XmCherry
FEATURES             Location/Qualifiers
     source          1..12104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     255..279
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      372..381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             384..404
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(417..458,459..461,462..2171,2172..2177,2178..2243,
                     2244..2246,2247..3659)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Neisseria meningitidis Type II 
                     CRISPR/Cas system"
                     /label=NmdCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D16A, D587A, 
                     H588A, and N611A"
                     /translation="AAFKPNPINYILGLAIGIASVGWAMVEIDEDENPICLIDLGVRVF
                     ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRARRLLKREGVLQAADFDENGLIKS
                     LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAD
                     NAHALQTGDFRTPAELALNKFEKESGHIRNQRGDYSHTFSRKDLQAELILLFEKQKEFG
                     NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
                     LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
                     AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
                     DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
                     EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
                     IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
                     LNEKGYVEIAAALPFSRTWDDSFNNKVLVLGSEAQNKGNQTPYEYFNGKDNSREWQEFK
                     ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
                     FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
                     GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTPEKLRTLLAEK
                     LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
                     EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
                     GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
                     IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
                     IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     regulatory      3246..3255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3678..3698
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3711..4418
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             4437..5144
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             5163..5870
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    5892..6480
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5945..5965)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6363..6374)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6555..6788
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6907..6925)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6993..7097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7098..7166,7167..7958)
                     /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(7316..7335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8129..8717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8618..8637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8871..8888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9128..9149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9164..9169,9170..9187,9188..9194)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9202..9218
                     /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     9207..9229
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9226..9242
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9226..9242
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9292..9311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9503..10136
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10183..10308
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10805..11038
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11223..11267
                     /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    11560..11677
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        11801..12104
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
      241 agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      301 tgttttgacc tccatagaag acaccgggac cgatccagcc taccggttaa ggtgaacgcg
      361 tgcaggctgg cgccaccatg gcccctaaaa agaaacgcaa ggtcgaattc ggaagcgccg
      421 ccttcaagcc caaccccatc aactacatcc tgggcctggc catcggcatc gccagcgtgg
      481 gctgggccat ggtggagatc gacgaggacg agaaccccat ctgcctgatc gacctgggtg
      541 tgcgcgtgtt cgagcgcgct gaggtgccca agactggtga cagtctggct atggctcgcc
      601 ggcttgctcg ctctgttcgg cgccttactc gccggcgcgc tcaccgcctt ctgcgcgctc
      661 gccgcctgct gaagcgcgag ggtgtgctgc aggctgccga cttcgacgag aacggcctga
      721 tcaagagcct gcccaacact ccttggcagc tgcgcgctgc cgctctggac cgcaagctga
      781 ctcctctgga gtggagcgcc gtgctgctgc acctgatcaa gcaccgcggc tacctgagcc
      841 agcgcaagaa cgagggcgag accgccgaca aggagctggg tgctctgctg aagggcgtgg
      901 ccgacaacgc ccacgccctg cagactggtg acttccgcac tcctgctgag ctggccctga
      961 acaagttcga gaaggagagc ggccacatcc gcaaccagcg cggcgactac agccacacct
     1021 tcagccgcaa ggacctgcag gccgagctga tcctgctgtt cgagaagcag aaggagttcg
     1081 gcaaccccca cgtgagcggc ggcctgaagg agggcatcga gaccctgctg atgacccagc
     1141 gccccgccct gagcggcgac gccgtgcaga agatgctggg ccactgcacc ttcgagccag
     1201 ccgagcccaa ggccgccaag aacacctaca ccgccgagcg cttcatctgg ctgaccaagc
     1261 tgaacaacct gcgcatcctg gagcagggca gcgagcgccc cctgaccgac accgagcgcg
     1321 ccaccctgat ggacgagccc taccgcaaga gcaagctgac ctacgcccag gcccgcaagc
     1381 tgctgggtct ggaggacacc gccttcttca agggcctgcg ctacggcaag gacaacgccg
     1441 aggccagcac cctgatggag atgaaggcct accacgccat cagccgcgcc ctggagaagg
     1501 agggcctgaa ggacaagaag agtcctctga acctgagccc cgagctgcag gacgagatcg
     1561 gcaccgcctt cagcctgttc aagaccgacg aggacatcac cggccgcctg aaggaccgca
     1621 tccagcccga gatcctggag gccctgctga agcacatcag cttcgacaag ttcgtgcaga
     1681 tcagcctgaa ggccctgcgc cgcatcgtgc ccctgatgga gcagggcaag cgctacgacg
     1741 aggcctgcgc cgagatctac ggcgaccact acggcaagaa gaacaccgag gagaagatct
     1801 acctgcctcc tatccccgcc gacgagatcc gcaaccccgt ggtgctgcgc gccctgagcc
     1861 aggcccgcaa ggtgatcaac ggcgtggtgc gccgctacgg cagccccgcc cgcatccaca
     1921 tcgagaccgc ccgcgaggtg ggcaagagct tcaaggaccg caaggagatc gagaagcgcc
     1981 aggaggagaa ccgcaaggac cgcgagaagg ccgccgccaa gttccgcgag tacttcccca
     2041 acttcgtggg cgagcccaag agcaaggaca tcctgaagct gcgcctgtac gagcagcagc
     2101 acggcaagtg cctgtacagc ggcaaggaga tcaacctggg ccgcctgaac gagaagggct
     2161 acgtggagat cgccgctgcc ctgcccttca gccgcacctg ggacgacagc ttcaacaaca
     2221 aggtgctggt gctgggcagc gaggctcaga acaagggcaa ccagaccccc tacgagtact
     2281 tcaacggcaa ggacaacagc cgcgagtggc aggagttcaa ggcccgcgtg gagaccagcc
     2341 gcttcccccg cagcaagaag cagcgcatcc tgctgcagaa gttcgacgag gacggcttca
     2401 aggagcgcaa cctgaacgac acccgctacg tgaaccgctt cctgtgccag ttcgtggccg
     2461 accgcatgcg cctgaccggc aagggcaaga agcgcgtgtt cgccagcaac ggccagatca
     2521 ccaacctgct gcgcggcttc tggggcctgc gcaaggtgcg cgccgagaac gaccgccacc
     2581 acgccctgga cgccgtggtg gtggcctgca gcaccgtggc catgcagcag aagatcaccc
     2641 gcttcgtgcg ctacaaggag atgaacgcct tcgacggtaa aaccatcgac aaggagaccg
     2701 gcgaggtgct gcaccagaag acccacttcc cccagccctg ggagttcttc gcccaggagg
     2761 tgatgatccg cgtgttcggc aagcccgacg gcaagcccga gttcgaggag gccgacaccc
     2821 ccgagaagct gcgcaccctg ctggccgaga agctgagcag ccgccctgag gccgtgcacg
     2881 agtacgtgac tcctctgttc gtgagccgcg cccccaaccg caagatgagc ggtcagggtc
     2941 acatggagac cgtgaagagc gccaagcgcc tggacgaggg cgtgagcgtg ctgcgcgtgc
     3001 ccctgaccca gctgaagctg aaggacctgg agaagatggt gaaccgcgag cgcgagccca
     3061 agctgtacga ggccctgaag gcccgcctgg aggcccacaa ggacgacccc gccaaggcct
     3121 tcgccgagcc cttctacaag tacgacaagg ccggcaaccg cacccagcag gtgaaggccg
     3181 tgcgcgtgga gcaggtgcag aagaccggcg tgtgggtgcg caaccacaac ggcatcgccg
     3241 acaacgccac catggtgcgc gtggacgtgt tcgagaaggg cgacaagtac tacctggtgc
     3301 ccatctacag ctggcaggtg gccaagggca tcctgcccga ccgcgccgtg gtgcagggca
     3361 aggacgagga ggactggcag ctgatcgacg acagcttcaa cttcaagttc agcctgcacc
     3421 ccaacgacct ggtggaggtg atcaccaaga aggcccgcat gttcggctac ttcgccagct
     3481 gccaccgcgg caccggcaac atcaacatcc gcatccacga cctggaccac aagatcggca
     3541 agaacggcat cctggagggc atcggcgtga agaccgccct gagcttccag aagtaccaga
     3601 tcgacgagct gggcaaggag atccgcccct gccgcctgaa gaagcgccct cctgtgcgcg
     3661 gctctactag tggctctccc aagaagaaga ggaaggtagg atccggaagt atggtgagca
     3721 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     3781 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     3841 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     3901 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     3961 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     4021 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc actgcaggac ggcgagttca
     4081 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     4141 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     4201 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     4261 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     4321 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgctgagg
     4381 gccgccactc caccggcggc atggacgagc tgtacaaggg tagcatcgat ggtagcatgg
     4441 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     4501 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4561 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     4621 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     4681 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     4741 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctcactg caggacggcg
     4801 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     4861 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     4921 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     4981 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     5041 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     5101 ctgagggccg ccactccacc ggcggcatgg acgagctgta caagtctggc ggccgctctg
     5161 gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     5221 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     5281 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     5341 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     5401 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     5461 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tcactgcagg
     5521 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     5581 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     5641 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     5701 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     5761 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     5821 aacgcgctga gggccgccac tccaccggcg gcatggacga gctgtacaag ctcgagtagg
     5881 gtaatctaga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5941 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6001 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6061 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6121 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6181 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6241 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     6301 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6361 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6421 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6481 ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6541 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     6601 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6661 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6721 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6781 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6841 aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat tcttgaagac
     6901 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6961 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7021 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7081 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7141 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7201 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7261 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7321 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     7381 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7441 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     7501 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     7561 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7621 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7681 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7741 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7801 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7861 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7921 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7981 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8041 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8101 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     8161 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8221 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     8281 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     8341 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     8401 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     8461 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     8521 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     8581 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     8641 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     8701 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     8761 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     8821 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     8881 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     8941 ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc
     9001 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     9061 ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     9121 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     9181 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     9241 acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     9301 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     9361 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     9421 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
     9481 tatgtactac aaacttagta gttggaaggg ctaattcact cccaaagaag acaagatatc
     9541 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     9601 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     9661 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     9721 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     9781 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     9841 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     9901 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     9961 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    10021 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    10081 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
    10141 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
    10201 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
    10261 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
    10321 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
    10381 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
    10441 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
    10501 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
    10561 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
    10621 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
    10681 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
    10741 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
    10801 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
    10861 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
    10921 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
    10981 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
    11041 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
    11101 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
    11161 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
    11221 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
    11281 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
    11341 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
    11401 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
    11461 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
    11521 ggtatcgccg aattgacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
    11581 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
    11641 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
    11701 agatccagtt tggactagcg atgctagctg atgcgggccc tagttattaa tagtaatcaa
    11761 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
    11821 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
    11881 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
    11941 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
    12001 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
    12061 ctacttggca gtacatctac gtattagtca tcgctattac catg
//
