LOCUS       sequence-176333-        9204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses St chimeric gRNA and human codon-optimized StCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9204)
  AUTHORS   Toth E, Weinhardt N, Bencsura P, Huszar K, Kulcsar PI, Talas A, 
            Fodor E, Welker E
  TITLE     Cpf1 nucleases demonstrate robust activity to induce DNA 
            modification by exploiting homology directed repair pathways in 
            mammalian cells.
  JOURNAL   Biol Direct. 2016 Sep 14;11:46. doi: 10.1186/s13062-016-0147-0.
  PUBMED    27630115
REFERENCE   2  (bases 1 to 9204)
  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"; journalName: "Biol 
            Direct."; date: "2016-09-14"; volume: "11:46. doi"; pages: " 
            10.1186/s13062-016-0147-0"
COMMENT     Sequence Label: pTE4254
FEATURES             Location/Qualifiers
     source          1..9204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        18..397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        398..601
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     552..572
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     598..622
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             716..4078
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus 
                     thermophilus Type II CRISPR/Cas system (Esvelt et al., 
                     2013)"
                     /label=St1Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MSDLVLGLDIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLV
                     RRTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEE
                     LFIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYG
                     QLRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGK
                     RKYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLL
                     NDLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRID
                     KSGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADG
                     SFSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKT
                     TSSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNED
                     DEKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERC
                     LYTGKTISIHDLINNSNQFEVDHILPLSITFDDSLANKVLVYATANQEKGQRTPYQALD
                     SMDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTLYASRVV
                     LNALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNL
                     WKKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSY
                     QVDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSK
                     FLMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGP
                     EIKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQ
                     FDKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRT
                     MPKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQ
                     HIIKNEGDKPKLDF"
     CDS             4091..4111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4167..4186)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4211..4259
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4461..4889
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4548..4567)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4758..4779
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4898..4918)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4903..5232
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5083..5218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5145..5164
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5299..6093
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5353..5372)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5963..5982
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6269..6390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6306..6325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6360..6379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6439..6455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6439..6455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6452..6474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6463..6479
                     /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(6487..6493,6494..6511,6512..6517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6532..6553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6670..6687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6841..7429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6921..6940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7600..8391,8392..8460))
                     /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     8223..8242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8461..8565)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8640..8659)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_RNA        complement(8776..8900)
                     /label=St1 gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus 
                     thermophilus CRISPR/Cas9 system (Esvelt et al., 2013)"
     promoter        complement(8921..9169)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(8979..8998)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(9149..9169)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 atgctagttg gcgcgttgac attgattatt gactagttat taatagtaat caattacggg
       61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      361 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      421 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      481 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      541 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      601 tcgtttagtg aaccgtcaga tcgcctggga tccccatcca cgctgttttg acctccatag
      661 aagacaccgg gaccgatcca gcctccggac tctagaggat cgaacccttg ccaccatgag
      721 cgacctggtg ctgggcctgg acatcggcat cggcagcgtg ggcgtgggca tcctgaacaa
      781 ggtgaccggc gagatcatcc acaagaacag tcgcatcttc cctgctgctc aggctgagaa
      841 caacctggtg cgccgcacca accgccaggg tcgccggctt gctcgccgca agaagcaccg
      901 gcgcgtgcgc ctgaaccgcc tgttcgagga gagcggcctg atcaccgact tcaccaagat
      961 cagcatcaac ctgaacccct accagctgcg cgtgaagggc ctgaccgacg agctgagcaa
     1021 cgaggagctg ttcatcgccc tgaagaacat ggtgaagcac cgcggcatca gctacctgga
     1081 cgacgccagc gacgacggca acagcagcgt gggcgactac gcccagatcg tgaaggagaa
     1141 cagcaagcag ctggagacca agacccccgg ccagatccag ctggagcgct accagaccta
     1201 cggccagctg cgcggcgact tcaccgtgga gaaggacggc aagaagcacc gcctgatcaa
     1261 cgtgttcccc accagcgcct accgcagcga ggccctgcgc atcctgcaga cccagcagga
     1321 gttcaacccc cagatcaccg acgagttcat caaccgctac ctggagatcc tgaccggcaa
     1381 gcgcaagtac taccacggcc ccggcaacga gaagagccgc accgactacg gccgctaccg
     1441 caccagcggc gagaccctgg acaacatctt cggcatcctg atcggcaagt gcaccttcta
     1501 ccccgacgag ttccgcgccg ccaaggccag ctacaccgcc caggagttca acctgctgaa
     1561 cgacctgaac aacctgaccg tgcccaccga gaccaagaag ctgagcaagg agcagaagaa
     1621 ccagatcatc aactacgtga agaacgagaa ggccatgggc cccgccaagc tgttcaagta
     1681 catcgccaag ctgctgagct gcgacgtggc cgacatcaag ggctaccgca tcgacaagag
     1741 cggcaaggcc gagatccaca ccttcgaggc ctaccgcaag atgaagaccc tggagaccct
     1801 ggacatcgag cagatggacc gagagaccct ggacaagctg gcctacgtgc tgaccctgaa
     1861 caccgagcgc gagggcatcc aggaggccct ggagcacgag ttcgccgacg gcagcttcag
     1921 ccagaagcag gtggacgagc tggtgcagtt ccgcaaggcc aacagcagca tcttcggcaa
     1981 gggctggcac aacttcagcg tgaagctgat gatggagctg atccccgagc tgtacgagac
     2041 cagcgaggag cagatgacca tcctgacccg cctgggcaag cagaagacca ccagcagcag
     2101 caacaagacc aagtacatcg acgagaagct gctgaccgag gagatctaca accccgtggt
     2161 ggccaagagc gtgcgccagg ccatcaagat cgtgaacgcc gccatcaagg agtacggcga
     2221 cttcgacaac atcgtgatcg agatggcccg cgagaccaac gaggacgacg agaagaaggc
     2281 catccagaag atccagaagg ccaacaagga cgagaaggac gccgccatgc tgaaggccgc
     2341 caaccagtac aacggcaagg ccgagctgcc ccacagcgtg ttccacggcc acaagcagct
     2401 ggccaccaag atccgcctgt ggcaccagca gggcgagcgc tgcctgtaca ccggcaagac
     2461 catcagcatc cacgacctga tcaacaacag caaccagttc gaggtggacc acatcctgcc
     2521 cctgagcatc accttcgacg acagcctggc caacaaggtg ctggtgtacg ccaccgccaa
     2581 ccaggagaag ggccagcgca ccccctacca ggccctggac agcatggacg acgcctggag
     2641 cttccgcgag ctgaaggcct tcgtgcgcga gagcaagacc ctgagcaaca agaagaagga
     2701 gtacctgctg accgaggagg acatcagcaa gttcgacgtg cgcaagaagt tcatcgagcg
     2761 caacctggtg gacaccctct acgccagccg cgtggtgctg aacgccctgc aggagcactt
     2821 ccgcgcccac aagatcgaca ccaaggtgag cgtggtgcgc ggccagttca ccagccagct
     2881 gcgccgccac tggggcatcg agaagacccg cgacacctac caccaccacg ccgtggacgc
     2941 cctgatcatt gcggcttcta gccagctgaa cctgtggaag aagcagaaga acaccctggt
     3001 gagctacagc gaggaccagc tgctggacat cgagaccggc gagctgatca gcgacgacga
     3061 gtacaaggag agcgtgttca aggcccccta ccagcacttc gtggacaccc tgaagagcaa
     3121 ggagttcgag gacagcatcc tgttcagcta ccaggtggac agcaagttca accgcaagat
     3181 cagcgacgcc accatctacg ccacccgcca ggccaaggtg ggcaaggaca aggccgacga
     3241 gacctacgtg ctgggcaaga tcaaggacat ctacacccag gacggctacg acgccttcat
     3301 gaagatctac aagaaggaca agagcaagtt cctgatgtac cgccacgacc cccagacctt
     3361 cgagaaggtg atcgagccca tcctggagaa ctaccccaac aagcagatca acgataaagg
     3421 caaggaggtg ccctgcaacc ccttcctgaa gtacaaggag gagcacggct acatccgcaa
     3481 gtacagcaag aagggcaacg gccccgagat caagagcctg aagtactacg acagcaagct
     3541 gggcaaccac atcgacatca cccccaagga cagcaacaac aaggtggtgc tgcagagcgt
     3601 gagcccctgg cgcgccgacg tgtacttcaa caagaccacc ggcaagtacg agatcctggg
     3661 cctgaagtac gccgacctgc agtttgataa gggcaccggc acctacaaga tcagccagga
     3721 gaagtacaac gacatcaaga agaaggaggg cgtggacagc gacagcgagt tcaagttcac
     3781 cctgtacaag aacgaccttc tgctggtgaa ggacaccgag accaaggagc aacagctgtt
     3841 ccgcttcctg agccgcacca tgcccaagca gaagcactac gtggagctga agccctacga
     3901 caagcagaag ttcgagggcg gcgaggccct gatcaaggtg ctgggcaacg tggccaacag
     3961 cggccagtgc aagaagggcc tgggcaagag caacatcagc atctacaagg tgcgcaccga
     4021 cgtgctgggc aaccagcaca tcatcaagaa cgagggcgac aagcccaagt tggacttcag
     4081 cagggctgac cccaagaaga agaggaaggt gtgagcaggg ttcgatccct accggttagt
     4141 aatgagttta aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4201 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     4261 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4321 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4381 cccagggctc gcagccaacg tcggggcggc agaccctgcc atagcagatc tgcgcagctg
     4441 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4501 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4561 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4621 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4681 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4741 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4801 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4861 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4981 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5041 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5161 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5221 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5281 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5341 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5401 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5461 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5521 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5581 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5641 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5701 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5761 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5821 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5881 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5941 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6001 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6061 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     6121 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6181 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6241 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6361 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6421 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6481 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6601 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7261 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8581 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     8641 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     8701 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     8761 tgcttaagaa aaaaacaccc tgccataaaa tgacagggtg ttgatttcgg catgaagcct
     8821 tatctttgta gcttctgcaa gatttaagta actgtgtaag tttcgttgta cagttactta
     8881 aatcttgaga gtacaaaaac ggagacgggt acccgtctcc ggtgtttcgt cctttccaca
     8941 agatatataa agccaagaaa tcgaaatact ttcaagttac ggtaagcata tgatagtcca
     9001 ttttaaaaca taattttaaa actgcaaact acccaagaaa ttattacttt ctacgtcacg
     9061 tattttgtac taatatcttt gtgtttacag tcaaattaat tctaattatc tctctaacag
     9121 ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa taggccctct tcctgcccga
     9181 ccttagatcc gaatccatat cgat
//
