LOCUS       sequence-420015-        8650 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express gRNA against PRDM14 with puro and BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8650)
  AUTHORS   Ishikawa M, Sugino S, Masuda Y, Tarumoto Y, Seto Y, Taniyama N, 
            Wagai F, Yamauchi Y, Kojima Y, Kiryu H, Yusa K, Eiraku M, Mochizuki 
            A
  TITLE     RENGE infers gene regulatory networks using time-series single-cell 
            RNA-seq data with CRISPR perturbations.
  JOURNAL   Commun Biol. 2023 Dec 28;6(1):1290. doi: 10.1038/s42003-023-05594-4.
  PUBMED    38155269
REFERENCE   2  (bases 1 to 8650)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2023-12-28"; volume: "6(1):1290. doi"; pages: " 
            10.1038/s42003-023-05594-4"
COMMENT     Sequence Label: pKLV2-U6gRNA5(PRDM14_v32_7-6)-PGKpuroBFP-W
FEATURES             Location/Qualifiers
     source          1..8650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        243..441
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     396..416
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             459..639
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    686..811
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1310..1543
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1728..1772
                     /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    2070..2187
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2262..2510
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2262..2282
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2433..2452
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        2639..3138
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2674..2697)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3071..3090
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3178..3774
                     /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(3178..3197)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3674..3694
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3793..3846
                     /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             3847..4548
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(4608..4628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5026..5037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5489..5506)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5495..5719
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5745..5765)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5745..5763)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5784..5800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5784..5800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5797..5819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5808..5824
                     /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(5832..5838,5839..5856,5857..5862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5877..5898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6015..6032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6186..6774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6266..6285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6945..7736,7737..7805))
                     /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     7568..7587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7806..7910)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7936..8391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8073..8094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8285..8304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8518..8540
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8532..8549
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8533..8549
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8559..8578
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8559..8577
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
       61 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      121 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      181 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      241 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      301 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      361 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      421 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
      481 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      541 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      601 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      661 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      721 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      781 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
      841 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
      901 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
      961 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1021 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1081 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1141 acagcaagcg gccggccgcg ctgatcttca gacctggagg aggagatatg agggacaatt
     1201 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     1261 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     1321 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     1381 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     1441 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     1501 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     1561 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     1621 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     1681 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     1741 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     1801 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     1861 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     1921 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     1981 agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc
     2041 tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     2101 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     2161 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt
     2221 tggttagtac cgggccctac gcgtccaagg tcgggcagga agagggccta tttcccatga
     2281 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
     2341 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     2401 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     2461 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggcagcagcc
     2521 acgagtacgc gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc
     2581 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttggatcca attctaccgg
     2641 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
     2701 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
     2761 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
     2821 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
     2881 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
     2941 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     3001 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     3061 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     3121 tcatctccgg gcctttcgac ctgcatccat ctagatctcg agcagctgaa gcttaccatg
     3181 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     3241 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     3301 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     3361 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     3421 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     3481 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     3541 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     3601 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     3661 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     3721 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgccggcggc
     3781 gggtccggag gagagggcag aggaagtctc ctaacatgcg gtgacgtgga ggagaatcct
     3841 ggcccaatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     3901 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     3961 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     4021 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     4081 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaggac
     4141 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     4201 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     4261 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     4321 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
     4381 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     4441 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     4501 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattgagc ggccgctcaa
     4561 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4621 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4681 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4741 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4801 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4861 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4921 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4981 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5041 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tctacgcctt
     5101 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcaggtacc tttaagacca
     5161 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     5221 gggctaattc actcccaaag aagtcaagat ctgctttttg cctgtactgg gtctctctgg
     5281 ttagaccaga gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5341 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5401 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5461 tctctagcag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     5521 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     5581 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     5641 ggtggggtgg ggcaggacag caagggggag gattgggaag tcaatagcag gcatgctggg
     5701 gatgcggtgg gctctatggg cggccgttaa ttaagctttt gttcccttta gtgagggtta
     5761 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5821 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     5881 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5941 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6001 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6061 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7261 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7561 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7921 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     7981 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     8041 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     8101 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     8161 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     8221 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     8281 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     8341 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     8401 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     8461 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     8521 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
     8581 aattgactag tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     8641 gcccattgac
//
