LOCUS       sequence-485524-        9938 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  Dual sgRNA targeting CTRL expressing BFP with Puromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9938)
  AUTHORS   Thayer JA, Petersen JD, Huang X, Hawrot J, Ramos DM, Sekine S, Li Y,
            Ward ME, Narendra DP
  TITLE     Novel reporter of the PINK1-Parkin mitophagy pathway identifies its 
            damage sensor in the import gate.
  JOURNAL   bioRxiv [Preprint]. 2025 Feb 20:2025.02.19.639160. doi: 
            10.1101/2025.02.19.639160.
  PUBMED    40027798
REFERENCE   2  (bases 1 to 9938)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Feb 20:2025.02.19.639160. doi: 
            10.1101/2025.02.19.639160."
COMMENT     Sequence Label: pLenti_DualsgRNA_CTRL_Puro_T2A_BFP2
FEATURES             Location/Qualifiers
     source          1..9938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             35..733
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     protein_bind    complement(753..786)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    842..1430
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(895..915)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1313..1324)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1433..1449)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1434..1450)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1959..2139
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2201..2789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2281..2300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2960..3751,3752..3820))
                     /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     3583..3602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3821..3925)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4000..4019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4191..4570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4572..4770
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4725..4745
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4788..4968
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5015..5140
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5637..5870
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6055..6099
                     /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    6397..6514
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(6767..6800)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     6804..6824
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     7049..7069
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7220..7239
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        8111..9289
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8342..9280
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     9237..9257
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             9301..9897
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(9301..9320)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9797..9817
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             join(9907..9938,1..22)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
ORIGIN
        1 cgtcgaggag aatcctggcc caccggtcgc caccagcgag ctgattaagg agaacatgca
       61 catgaagctg tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg
      121 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc
      181 tctccccttc gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat
      241 caaccacacc cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg
      301 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
      361 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg
      421 ccctgtgatg cagaagaaaa cactcggctg ggaggccttc accgagaccc tgtaccccgc
      481 tgacggcggc ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct
      541 gatcgcaaac atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc
      601 tggcgtctac tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta
      661 cgtcgagcag cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca
      721 caagcttaat taagaattcg tcgagggacc taataacttc gtatagcata cattatacga
      781 agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga
      841 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
      901 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
      961 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     1021 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     1081 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     1141 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1201 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     1261 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     1321 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     1381 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     1441 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     1501 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     1561 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     1621 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     1681 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     1741 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     1801 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     1861 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     1921 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
     1981 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     2041 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     2101 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc
     2161 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2221 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2281 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2341 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2401 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2461 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2521 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2581 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2641 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2701 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2761 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2821 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2881 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2941 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3001 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3061 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3121 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3181 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3241 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3301 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3361 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3421 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3481 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3541 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3601 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3661 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3721 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3781 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3841 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3901 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     3961 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     4021 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     4081 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     4141 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     4201 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     4261 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     4321 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     4381 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     4441 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     4501 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     4561 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     4621 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     4681 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     4741 gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt
     4801 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     4861 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     4921 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     4981 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     5041 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     5101 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     5161 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     5221 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     5281 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     5341 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     5401 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     5461 gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg
     5521 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     5581 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     5641 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     5701 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     5761 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     5821 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     5881 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     5941 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     6001 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     6061 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     6121 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     6181 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     6241 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     6301 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     6361 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     6421 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     6481 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     6541 tccagtttgg ttagtaccgg gcccgctcta gagatccgac gcgccatctc taggcccgcg
     6601 ccggccccct cgcacggact tgtgggagaa gctcggctac tcccctgccc cggttaattt
     6661 gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac agataatctg
     6721 ttctttttaa tactagctac attttacatg ataggcttgg atttctataa cttcgtatag
     6781 catacattat acgaagttat aaacagcaca aaaggaaact caccctaact gtaaagtaat
     6841 tgtgtgtttt gagactataa gtatcccttg gagaaccacc ttgttgggcc gtgggcaaca
     6901 ctgtatgttt cagagctaag cacaagagtg catagcaagt tgaaataagg ctagtccgtt
     6961 tacaacttga aaaagtggca cccgagtcgg gtgctttttt tgctcgaatc tacactcagc
     7021 tatggcgcgc cccaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt
     7081 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
     7141 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     7201 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     7261 cttggcttta tatatcttgt ggaaagccag aaacatggcg ttggcactta gcgcagcgtt
     7321 taagagctaa gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg
     7381 aaaaagtggc accgagtcgg tgcttttttt ctcgagtact aggatccatt aggcggccgc
     7441 cgggaggtgg tccctgcagt tacgccaatg ataacccccg ccagaaaaat cttagtagcc
     7501 ttcccttttt gttttccgtg ccccaactcg gcggattgac tcggcccctt ccggaaacac
     7561 ccgaatcaac ttctagtcaa attattgttc acgccgcaat gacccacccc tggcccgcgt
     7621 ctgtggaact gacccctggt gtacaggaga gttcgctgct gaaagtggtc ccaaaggggt
     7681 actagttttt aagctcccaa ctccccctcc cccagcgtct ggaggattcc acaccctcgc
     7741 accgcagggg cgaggaagtg ggcggagtcc ggttttggcg ccagccgctg aggctgccaa
     7801 gcagaaaagc caccgctgag gagactccgg tcactgtcct cgccccgcct cccccttccc
     7861 tccccttggg gaccaccggg cgccacgccg cgaacggtaa gtgccgcggt cgtcggcgcc
     7921 tccgccctcc ccctagggcc ccaattccca gcgggcgcgg cgcgcggccc ctccccccgc
     7981 cgggcgcgcg cccgctgccc cgcccttcgt ggccgcccgg cgtgggcggt gccacccctc
     8041 cccccggcgg ccccgcgcgc agctcccggc tccctccccc ttcggatgtg gcttgagctg
     8101 taggcgcgga gatgcatgtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     8161 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     8221 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     8281 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     8341 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     8401 gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     8461 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     8521 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     8581 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     8641 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     8701 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     8761 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     8821 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     8881 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     8941 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     9001 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     9061 gcacctcgat tagttctcgc gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     9121 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     9181 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     9241 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag ctagcccacc
     9301 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     9361 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     9421 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     9481 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     9541 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     9601 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     9661 cccgcgtggt tcctggccac cgtcggcgtg tcgcccgacc accagggcaa gggtctgggc
     9721 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     9781 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     9841 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgccgga
     9901 tcgggagagg gcagaggaag tctgctaaca tgcggtga
//
