LOCUS       sequence-341912-        9003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for easy cloning of a sgRNA sequence. The Plasmid contain a 
            tRNA promoter to facilitate the expression of any sgRNA sequence and
            also expresses eSpCas9(1.1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9003)
  AUTHORS   Perez-Leal O, Nixon-Abell J, Barrero CA, Gordon JC, Oesterling J, 
            Rico MC
  TITLE     Multiplex Gene Tagging with CRISPR-Cas9 for Live-Cell Microscopy and
            Application to Study the Role of SARS-CoV-2 Proteins in Autophagy, 
            Mitochondrial Dynamics, and Cell Growth.
  JOURNAL   CRISPR J. 2021 Nov 30. doi: 10.1089/crispr.2021.0041.
  PUBMED    34847745
REFERENCE   2  (bases 1 to 9003)
  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: "CRISPR J. 
            2021 Nov 30. doi: 10.1089/crispr.2021.0041."
COMMENT     Sequence Label: FASTHDR-tRNApromo-sgRNA-espCas91_1
FEATURES             Location/Qualifiers
     source          1..9003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      91..679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     580..599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1150..1455
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1334..1353
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1506..1581
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1678..1963
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1965..2242
                     /label=chicken beta-actin promoter
     intron          2243..2470
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2482..2491
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2491..2556
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2563..2583
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2608..5145,5146..5148,5149..5610,5611..5613,5614..5781,
                     5782..5784,5785..6708)
                     /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             6709..6756
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6784..6801)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6790..6997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7006..7146
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7006..7135
                     /label=AAV2 ITR
     rep_origin      7221..7676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7308..7327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7518..7539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7693..7712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7812..7834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7872..7890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7958..8062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8063..8131,8132..8923)
                     /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(8281..8300)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
       61 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      301 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      361 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      421 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      481 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      541 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      601 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      661 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      721 ctggcctttt gctcacatgt ggttccatgg tgtaatggtt agcactctgg actctgaatc
      781 cagcgatccg agttcaaatc tcggtggaac ctgggtcttc ggcttactaa aagccagata
      841 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
      901 ccgaagtatg tcaaaaagag gtgtgctatg aagcagcgta ttacagtgac agttgacagc
      961 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     1021 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     1081 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     1141 actggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     1201 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     1261 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     1321 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     1381 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     1441 ttctggggaa tataaatgtc aggctccgtt atacacagcc agtctgcagg tcgacgagaa
     1501 gacctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     1561 aaagtggcac cgagtcggtg cttttttgtt ttagagctag gattagcaag ttcgagtaag
     1621 gctcctgagt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag aggtacccgt
     1681 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     1741 gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     1801 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     1861 caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat gggactttcc
     1921 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
     1981 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
     2041 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
     2101 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
     2161 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     2221 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
     2281 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     2341 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
     2401 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
     2461 tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag
     2521 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
     2581 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
     2641 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     2701 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
     2761 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     2821 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     2881 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     2941 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     3001 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     3061 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     3121 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     3181 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     3241 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     3301 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     3361 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     3421 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     3481 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     3541 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     3601 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     3661 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     3721 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     3781 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     3841 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     3901 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     3961 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     4021 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     4081 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     4141 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     4201 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     4261 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     4321 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     4381 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     4441 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     4501 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     4561 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     4621 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     4681 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     4741 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     4801 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     4861 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     4921 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     4981 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     5041 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     5101 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctggccga cgactccatc
     5161 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     5221 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     5281 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     5341 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     5401 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     5461 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     5521 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     5581 gtgggaaccg ccctgatcaa aaagtaccct gcgctggaaa gcgagttcgt gtacggcgac
     5641 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     5701 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     5761 gccaacggcg agatccggaa ggcgcctctg atcgagacaa acggcgaaac cggggagatc
     5821 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     5881 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     5941 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     6001 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     6061 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     6121 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     6181 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     6241 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     6301 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     6361 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     6421 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     6481 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     6541 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     6601 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     6661 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     6721 gccacgaaaa aggccggcca ggcaaaaaag aaaaagtaag aattcctaga gctcgctgat
     6781 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6841 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     6901 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     6961 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga
     7021 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     7081 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     7141 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     7201 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     7261 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     7321 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     7381 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     7441 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7501 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     7561 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     7621 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     7681 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     7741 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     7801 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     7861 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7921 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7981 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     8041 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     8101 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     8161 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     8221 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     8281 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     8341 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     8401 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     8461 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     8521 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     8581 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     8641 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     8701 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8761 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     8821 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8881 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8941 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     9001 gat
//
