LOCUS       sequence-347396-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses spCas9 and gRNA sequence targeting TRERF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  TITLE     ENCODE collection: CRISPR constructs
REFERENCE   2  (bases 1 to 8508)
  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"
COMMENT     Sequence Label: pTRERF1.1.0-gDNA
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..76
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        173..458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          738..965
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      977..986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             986..1051
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1058..1078
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1103..3640,3641..3643,3644..4105,4106..4108,4109..4276,
                     4277..4279,4280..5203)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5204..5251
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5279..5296)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5285..5492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5501..5641
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5501..5630
                     /label=AAV2 ITR
     rep_origin      5716..6171
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5803..5822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6013..6034
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6188..6207)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6307..6329
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6367..6385)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6453..6557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6558..6626,6627..7418)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6776..6795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7589..8177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8078..8097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8239..8487
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8239..8259
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8410..8429
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
       61 ggcaccgagt cggtgctttt ttgttttaga gctagaaata gcaagttaaa ataaggctag
      121 tccgttttta gcgcgtgcgc caattctgca gacaaatggc tctagaggta cccgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      361 acggtaaatg gcccgcctgg cattgtgccc agtacatgac cttatgggac tttcctactt
      421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      481 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      541 aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgccr ggsggggggg
      601 gggggggsgg ggggsggggs ggggsgrggs ggrgaggkgc ggcggcagcc aatcagagcg
      661 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      721 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      781 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      841 gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg tttaagggat
      901 ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa atcacttttt
      961 ttcaggttgg accggtgcca ccatggacta taaggaccac gacggagact acaaggatca
     1021 tgatattgat tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg
     1081 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     1141 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     1201 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     1261 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     1321 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     1381 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     1441 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     1501 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     1561 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     1621 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     1681 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     1741 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     1801 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     1861 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     1921 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     1981 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     2041 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     2101 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     2161 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     2221 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     2281 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     2341 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     2401 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     2461 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     2521 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     2581 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     2641 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     2701 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     2761 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     2821 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     2881 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     2941 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     3001 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     3061 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     3121 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     3181 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     3241 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     3301 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     3361 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     3421 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     3481 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     3541 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     3601 cgactacgat gtggaccata tcgtgcctca gagctttctg gccgacgact ccatcgacaa
     3661 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     3721 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     3781 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     3841 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     3901 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     3961 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     4021 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     4081 aaccgccctg atcaaaaagt accctgcgct ggaaagcgag ttcgtgtacg gcgactacaa
     4141 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     4201 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     4261 cggcgagatc cggaaggcgc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     4321 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     4381 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     4441 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     4501 cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     4561 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     4621 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     4681 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     4741 ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     4801 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     4861 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     4921 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     4981 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     5041 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     5101 gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     5161 cctgtacgag acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac
     5221 gaaaaaggcc ggccaggcaa aaaagaaaaa gtaagaattc ctagagctcg ctgatcagcc
     5281 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     5341 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     5401 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     5461 gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag
     5521 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     5581 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     5641 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     5701 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5761 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     5821 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5881 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5941 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6001 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     6061 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     6121 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     6181 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     6241 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6301 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     6361 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6421 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6481 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6541 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6601 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6661 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6721 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6781 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6841 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6901 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6961 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     7021 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     7081 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     7141 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7201 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7261 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7321 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7381 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7441 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7501 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7561 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7621 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7681 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7741 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7801 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7861 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7921 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7981 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     8041 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     8101 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     8161 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     8221 ggccttttgc tcacatgtga gggcctattt cccatgattc cttcatattt gcatatacga
     8281 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
     8341 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     8401 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     8461 tatatcttgt ggaaaggacg aaacaccgac tgtctctaag tgacacac
//
