LOCUS       sequence-373474-       13319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged PlmdCas12e fused to VPR (VP64-p65-Rta).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13319)
  AUTHORS   Escobar M, Li J, Patel A, Liu S, Xu Q, Hilton IB
  TITLE     Quantification of Genome Editing and Transcriptional Control 
            Capabilities Reveals Hierarchies among Diverse CRISPR/Cas Systems in
            Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Oct 21;11(10):3239-3250. doi: 
            10.1021/acssynbio.2c00156. Epub 2022 Sep 26.
  PUBMED    36162812
REFERENCE   2  (bases 1 to 13319)
  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: 
            "10.1021/acssynbio.2c00156"; journalName: "ACS Synth Biol"; date: 
            "2022-10-21- 21"; volume: "11"; issue: "10"; pages: "3239-3250"
COMMENT     Sequence Label: PlmdCas12e VPR
FEATURES             Location/Qualifiers
     source          1..13319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        2071..2146
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2208..2419
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5378..5425
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5426..5449
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    5492..5516
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5546..5695
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5720..5740
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6170..6526
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             6545..7114
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             7124..7180
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7181..7777
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7674..7694
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7793..8381
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7846..7866)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8264..8275)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8453..8686
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8712..8729)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8718..8942
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8988..9416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9075..9094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9285..9306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9425..9445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9430..9759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9610..9745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9672..9691
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9807..9854
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9873..10247
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10377..10498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10414..10433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10468..10487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10547..10563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10547..10563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10560..10582)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10571..10587
                     /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)."
     promoter        complement(join(10595..10601,10602..10619,10620..10625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10640..10661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10778..10795)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10949..11537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11029..11048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11708..12499,12500..12568))
                     /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     12331..12350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12569..12673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12748..12767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12939..13318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg atatcactag tgttccatta actgcgctaa aaataatttt taaatctttt
     2041 ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc aagttaaaat
     2101 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc
     2161 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     2221 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     2281 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2341 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2401 gtttgccgcc agaacacagg accggttcta gagcgctgcc accatgcaag agatcaagag
     2461 aatcaacaag atcagaagga gactggtcaa ggacagcaac acaaagaagg ccggcaagac
     2521 aggccccatg aaaaccctgc tcgtcagagt gatgacccct gacctgagag agcggctgga
     2581 aaacctgaga aagaagcccg agaacatccc tcagcctatc agcaacacca gcagggccaa
     2641 cctgaacaag ctgctgaccg actacaccga gatgaagaaa gccatcctgc acgtgtactg
     2701 ggaagagttc cagaaagacc ccgtgggcct gatgagcaga gttgctcagc ccgctcctaa
     2761 gaacatcgac cagagaaagc tgatccccgt gaaggacggc aacgagagac tgacctctag
     2821 cggctttgcc tgcagccagt gttgccagcc tctgtacgtg tacaagctgg aacaagtgaa
     2881 cgacaagggc aagccccaca ccaactactt cggcagatgc aacgtgtccg agcacgagag
     2941 gctgatcctg ctgtctcctc acaagcccga ggccaacgat gagctggtca catacagcct
     3001 gggcaagttc ggacagagag ccctggactt ctacagcatc cacgtgacca gggagagcaa
     3061 tcaccctgtg aagcccctgg aacagatcgg cggcaatagc tgtgcctctg gacctgtggg
     3121 aaaagccctg agcgacgcct gtatgggagc cgtggcatcc ttcctgacca agtaccagga
     3181 catcatcctg gaacaccaga aagtgatcaa gaagaacgag aaaagactgg ccaacctcaa
     3241 ggatatcgcc agcgctaacg gcctggcctt tcctaagatc accctgcctc cacagcctca
     3301 caccaaagag ggcatcgagg cctacaacaa cgtggtggcc cagatcgtga tttgggtcaa
     3361 cctgaatctg tggcagaagc tgaagatcgg cagggacgaa gccaagccac tgcagagact
     3421 gaagggcttc cctagcttcc ctctggtgga aagacaggcc aatgaagtgg attggtggga
     3481 catggtctgc aacgtgaaga agctgatcaa cgagaagaaa gaggatggca aggttttctg
     3541 gcagaacctg gccggctaca agagacaaga agccctgctg ccttacctga gcagcgaaga
     3601 ggaccggaag aagggcaaga agttcgccag ataccagttc ggcgacctgc tgctgcacct
     3661 ggaaaagaag cacggcgagg actggggcaa agtgtacgat gaggcctggg agagaatcga
     3721 caagaaggtg gaaggcctga gcaagcacat taagctggaa gaggaaagaa ggagcgagga
     3781 cgcccaatct aaagccgctc tgaccgattg gctgagagcc aaggccagct ttgtgatcga
     3841 gggcctgaaa gaggccgaca aggacgagtt ctgcagatgc gagctgaagc tgcagaagtg
     3901 gtacggcgat ctgagaggca agcccttcgc cattgaggcc gagaacagca tcctggacat
     3961 cagcggcttc agcaagcagt acaactgcgc cttcatttgg cagaaagacg gcgtcaagaa
     4021 actgaacctg tacctgatca tcaattactt caaaggcggc aagctgcggt tcaagaagat
     4081 caaacccgag gccttcgagg ctaacagatt ctacaccgtg atcaacaaaa agtccggcga
     4141 gatcgtgccc atggaagtga acttcaactt cgacgacccc aacctgatta tcctgcctct
     4201 ggccttcggc aagagacagg gcagagagtt catctggaac gatctgctga gcctggaaac
     4261 cggctctctg aagctggcca atggcagagt gatcgagaaa accctgtaca acaggagaac
     4321 cagacaggac gagcctgctc tgtttgtggc cctgaccttc gagagaagag aggtgctgga
     4381 cagcagcaac atcaagccca tgaacctgat cggcatcgcc cggggcgaga atatccctgc
     4441 tgtgatcgcc ctgacagacc ctgaaggatg cccactgagc agattcaagg actccctggg
     4501 caaccctaca cacatcctga gaatcggcga gagctacaaa gagaagcaga ggacaatcca
     4561 ggccgccaaa gaggtggaac agagaagagc cggcggatac tctaggaagt acgccagcaa
     4621 ggccaagaat ctggccgacg acatggtccg aaacaccgcc agagatctgc tgtactacgc
     4681 cgtgacacag gacgccatgc tgatcttcgc caatctgagc agaggcttcg gccggcaggg
     4741 caagagaacc tttatggccg agaggcagta caccagaatg gaagattggc tcacagctaa
     4801 actggcctac gagggactgc ccagcaagac ctacctgtcc aaaacactgg cccagtatac
     4861 ctccaagacc tgcagcaatt gcggcttcac catcaccagc gccgactacg acagagtgct
     4921 ggaaaagctc aagaaaaccg ccaccggctg gatgaccacc atcaacggca aagagctgaa
     4981 ggttgagggc cagatcacct actacaacag gtacaagagg cagaacgtcg tgaaggatct
     5041 gagcgtggaa ctggacagac tgagcgaaga gagcgtgaac aacgacatca gcagctggac
     5101 aaagggcaga tcaggcgagg ctctgagcct gctgaagaag aggtttagcc acagacctgt
     5161 gcaagagaag ttcgtgtgcc tgaactgcgg cttcgagaca cacgccgccg aacaggctgc
     5221 cctgaacatt gccagaagct ggctgttcct gagaagccaa gagtacaaga agtaccagac
     5281 caacaagacc accggcaaca ccgacaagag ggcctttgtg gaaacctggc agagcttcta
     5341 cagaaaaaag ctgaaagaag tctggaagcc cgccgtgaag cgacctgccg ccacaaagaa
     5401 ggctggacag gctaagaaga agaaagatta caaagacgat gacgataagg gatcctcgcc
     5461 agggatccgt cgacttgacg cgttgatatc aacaagtttg tacaaaaaag caggctacaa
     5521 agaggccagc ggttccggac gggctgacgc attggacgat tttgatctgg atatgctggg
     5581 aagtgacgcc ctcgatgatt ttgaccttga catgcttggt tcggatgccc ttgatgactt
     5641 tgacctcgac atgctcggca gtgacgccct tgatgatttc gacctggaca tgctgattaa
     5701 ctctagaagt tccggatctc cgaaaaagaa acgcaaagtt ggtagccagt acctgcccga
     5761 caccgacgac cggcaccgga tcgaggaaaa gcggaagcgg acctacgaga cattcaagag
     5821 catcatgaag aagtccccct tcagcggccc caccgaccct agacctccac ctagaagaat
     5881 cgccgtgccc agcagatcca gcgccagcgt gccaaaacct gccccccagc cttacccctt
     5941 caccagcagc ctgagcacca tcaactacga cgagttccct accatggtgt tccccagcgg
     6001 ccagatctct caggcctctg ctctggctcc agcccctcct caggtgctgc ctcaggctcc
     6061 tgctcctgca ccagctccag ccatggtgtc tgcactggct caggcaccag cacccgtgcc
     6121 tgtgctggct cctggacctc cacaggctgt ggctccacca gcccctaaac ctacacaggc
     6181 cggcgagggc acactgtctg aagctctgct gcagctgcag ttcgacgacg aggatctggg
     6241 agccctgctg ggaaacagca ccgatcctgc cgtgttcacc gacctggcca gcgtggacaa
     6301 cagcgagttc cagcagctgc tgaaccaggg catccctgtg gcccctcaca ccaccgagcc
     6361 catgctgatg gaataccccg aggccatcac ccggctcgtg acaggcgctc agaggcctcc
     6421 tgatccagct cctgcccctc tgggagcacc aggcctgcct aatggactgc tgtctggcga
     6481 cgaggacttc agctctatcg ccgatatgga tttctcagcc ttgctgggct ctggcagcgg
     6541 cagccgggat tccagggaag ggatgttttt gccgaagcct gaggccggct ccgctattag
     6601 tgacgtgttt gagggccgcg aggtgtgcca gccaaaacga atccggccat ttcatcctcc
     6661 aggaagtcca tgggccaacc gcccactccc cgccagcctc gcaccaacac caaccggtcc
     6721 agtacatgag ccagtcgggt cactgacccc ggcaccagtc cctcagccac tggatccagc
     6781 gcccgcagtg actcccgagg ccagtcacct gttggaggat cccgatgaag agacgagcca
     6841 ggctgtcaaa gcccttcggg agatggccga tactgtgatt ccccagaagg aagaggctgc
     6901 aatctgtggc caaatggacc tttcccatcc gcccccaagg ggccatctgg atgagctgac
     6961 aaccacactt gagtccatga ccgaggatct gaacctggac tcacccctga ccccggaatt
     7021 gaacgagatt ctggatacct tcctgaacga cgagtgcctc ttgcatgcca tgcatatcag
     7081 cacaggactg tccatcttcg acacatctct gtttggatcc ggcgcaacaa acttctctct
     7141 gctgaaacaa gccggagatg tcgaagagaa tcctggaccg accgagtaca agcccacggt
     7201 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     7261 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     7321 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     7381 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     7441 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     7501 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     7561 cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     7621 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     7681 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     7741 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc
     7801 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     7861 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     7921 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     7981 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     8041 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     8101 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     8161 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     8221 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     8281 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     8341 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg
     8401 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     8461 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     8521 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     8581 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     8641 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
     8701 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     8761 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     8821 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     8881 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     8941 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     9001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     9061 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     9121 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     9181 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     9241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     9301 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     9361 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     9421 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     9481 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     9541 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     9601 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     9661 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     9721 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     9781 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     9841 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     9901 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     9961 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
    10021 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
    10081 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
    10141 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
    10201 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
    10261 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
    10321 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
    10381 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
    10441 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
    10501 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
    10561 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
    10621 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
    10681 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
    10741 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
    10801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
    10861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
    10921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
    10981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
    11041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
    11101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
    11161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
    11221 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
    11281 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
    11341 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
    11401 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
    11461 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
    11521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
    11581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
    11641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
    11701 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
    11761 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
    11821 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
    11881 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
    11941 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
    12001 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
    12061 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
    12121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    12181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    12241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    12301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    12361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    12421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    12481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    12541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
    12601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
    12661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
    12721 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
    12781 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
    12841 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
    12901 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
    12961 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
    13021 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
    13081 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
    13141 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
    13201 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
    13261 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgg
//
