LOCUS       sequence-214952-        8650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral gRNA plasmid targeting mouse Nelfcd , co-expression of 
            TagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8650)
  AUTHORS   Li M, Yu JSL, Tilgner K, Ong SH, Koike-Yusa H, Yusa K
  TITLE     Genome-wide CRISPR-KO Screen Uncovers mTORC1-Mediated Gsk3 
            Regulation in Naive Pluripotency Maintenance and Dissolution.
  JOURNAL   Cell Rep. 2018 Jul 10;24(2):489-502. doi: 
            10.1016/j.celrep.2018.06.027.
  PUBMED    29996108
REFERENCE   2  (bases 1 to 8650)
  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: 
            "10.1016/j.celrep.2018.06"; journalName: "Cell Rep"; date: 
            "2018-07-10- 10"; volume: "24"; issue: "2"; pages: "489-502"
COMMENT     Sequence Label: pKLV2-U6gRNA5(Nelfcd-g2)-PGKpuroBFP-W
FEATURES             Location/Qualifiers
     source          1..8650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        191..389
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     344..364
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             407..587
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    634..759
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1258..1491
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1676..1720
                     /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    2018..2135
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2210..2458
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2210..2230
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2381..2400
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        2587..3086
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2622..2645)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3019..3038
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3126..3722
                     /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(3126..3145)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3622..3642
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3741..3794
                     /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"
     CDS             3795..4496
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(4556..4576)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4974..4985)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5437..5454)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5443..5667
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5693..5713)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5693..5711)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5732..5748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5732..5748)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5745..5767)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5756..5772
                     /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(5780..5786,5787..5804,5805..5810))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5825..5846
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5963..5980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6134..6722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6214..6233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6893..7684,7685..7753))
                     /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     7516..7535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7754..7858)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7884..8339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8021..8042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8233..8252
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8466..8488
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8480..8497
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8481..8497
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8507..8526
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8507..8525
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
       61 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      121 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      181 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      241 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      301 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      361 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
      421 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
      481 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
      541 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
      601 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
      661 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
      721 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
      781 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
      841 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
      901 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
      961 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     1021 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     1081 accaccgcac agcaagcggc cggccgcgct gatcttcaga cctggaggag gagatatgag
     1141 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1201 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1261 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1321 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1381 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1441 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1501 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1561 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1621 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1681 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1741 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1801 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1861 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1921 aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc
     1981 gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     2041 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     2101 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     2161 atccagtttg gttagtaccg ggccctacgc gtccaaggtc gggcaggaag agggcctatt
     2221 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat
     2281 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     2341 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     2401 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt
     2461 ctgatgcggg gtaccaaggt ttaagagcta tgctggaaac agcatagcaa gtttaaataa
     2521 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tggatccaat
     2581 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     2641 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     2701 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
     2761 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
     2821 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
     2881 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
     2941 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
     3001 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
     3061 cctcttcctc atctccgggc ctttcgacct gcatccatct agatctcgag cagctgaagc
     3121 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     3181 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     3241 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     3301 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     3361 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     3421 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     3481 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     3541 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     3601 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     3661 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     3721 ccggcggcgg gtccggagga gagggcagag gaagtctcct aacatgcggt gacgtggagg
     3781 agaatcctgg cccaatgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
     3841 agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg
     3901 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     3961 acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca
     4021 tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
     4081 acgaggacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     4141 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     4201 aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag
     4261 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga
     4321 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     4381 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     4441 tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt aattgagcgg
     4501 ccgctcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4561 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4621 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4681 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4741 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4801 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4861 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     4921 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     4981 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5041 tacgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg caggtacctt
     5101 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5161 gactggaagg gctaattcac tcccaaagaa gtcaagatct gctttttgcc tgtactgggt
     5221 ctctctggtt agaccagagt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5281 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5341 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     5401 gtggaaaatc tctagcagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     5461 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5521 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5581 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagtc aatagcaggc
     5641 atgctgggga tgcggtgggc tctatgggcg gccgttaatt aagcttttgt tccctttagt
     5701 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5761 atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg
     5821 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5881 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5941 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6001 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6061 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6241 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6301 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6361 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6421 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6481 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6541 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6601 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6661 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6721 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6781 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6841 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6961 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7021 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7081 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7321 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7381 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7561 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7741 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7801 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7861 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     7921 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     7981 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     8041 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     8101 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     8161 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     8221 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     8281 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     8341 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     8401 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     8461 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     8521 atagggcgaa ttgactagta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     8581 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     8641 tttccattga
//
