LOCUS       sequence-419749-        9837 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral guide RNA expression plasmid for CRISPR activation with 
            puromycin/BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9837)
  AUTHORS   Yang L, Sheets TP, Feng Y, Yu G, Bajgain P, Hsu KS, So D, Seaman S, 
            Lee J, Lin L, Evans CN, Guest MR, Chari R, St Croix B
  TITLE     Uncovering receptor-ligand interactions using a high-avidity CRISPR 
            activation screening platform.
  JOURNAL   Sci Adv. 2024 Feb 16;10(7):eadj2445. doi: 10.1126/sciadv.adj2445. 
            Epub 2024 Feb 14.
  PUBMED    38354234
REFERENCE   2  (bases 1 to 9837)
  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: "Sci Adv."; 
            date: "2024-02-16"; volume: "10(7):eadj2445. doi"; pages: " 
            10.1126/sciadv.adj2445. Epub 2024 Feb 14"
COMMENT     Sequence Label: pGMC00029 (aka pRC0610)
FEATURES             Location/Qualifiers
     source          1..9837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2246..2494
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2246..2266
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2417..2436
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        2793..3096
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3098..3371
                     /label=chicken beta-actin promoter
     intron          3372..4388
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4312..4334
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4396..4415
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4422..5018
                     /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(4422..4441)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4918..4938
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5034..5090
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5091..5792
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    5805..6393
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5858..5878)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6276..6287)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6600..7233
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7362..7378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7362..7378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7375..7397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7386..7402
                     /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(7410..7416,7417..7434,7435..7440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7455..7476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7593..7610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7764..8352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7844..7863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8523..9314,9315..9383))
                     /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     9146..9165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9384..9488)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9536..9670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9573..9592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9627..9646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cattagtgaa cggatctcga cggtatcgat cacgagacta
     2221 gcctcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca
     2281 tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat
     2341 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     2401 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
     2461 ggctttatat atcttgtgga aaggacgaaa caccggagac gaccggtttt ttgcgtctct
     2521 gtttaagagc tatgctgcga atacgaggga gcagacgata tggcgtcgct cctctccacg
     2581 agagcatatg ggctccgtgg tctcgtacac catcagggta cgtatcagac accatcaggg
     2641 tctgctcgta ttcgcagcat agcaagttta aataaggcta gtccgttatc aacttgaaaa
     2701 agtggcaccg agtcggtgct ttttttgaat tctcgacctc gagacaaatg gcagtattcg
     2761 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2821 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2881 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2941 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     3001 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     3061 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     3121 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     3181 ttattttgtg cagcgatggg ggcggggggg gggggcgcgc gccaggcggg gcggggcggg
     3241 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     3301 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     3361 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
     3421 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
     3481 gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt
     3541 ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg
     3601 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg
     3661 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga
     3721 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg
     3781 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac
     3841 cccccctgca cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc
     3901 gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc
     3961 cgggcggggc ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc
     4021 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
     4081 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc
     4141 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa
     4201 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc
     4261 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     4321 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     4381 tcctacagct cctgggcaac gtgctggtta ttgtggccac catgaccgag tacaagccca
     4441 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4501 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4561 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4621 cggacgacgg cgccgccgtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4681 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     4741 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     4801 ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     4861 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     4921 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     4981 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgc tagcggcagc ggcgccacca
     5041 acttcagcct gctgaagcag gccggcgacg tggaggagaa tcctggaccg atgagcgagc
     5101 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact
     5161 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
     5221 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct
     5281 acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct
     5341 tccctgaggg cttcacatgg gagagagtca ccacatacga ggacgggggc gtgctgaccg
     5401 ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
     5461 tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca
     5521 ccgaaacact gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc
     5581 tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac atatagatcc aagaaacccg
     5641 ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg
     5701 aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc
     5761 tccctagcaa actggggcac aagcttaatt gaagcgtctg gaacaatcaa cctctggatt
     5821 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     5881 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     5941 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6001 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6061 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6121 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6181 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     6241 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     6301 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     6361 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct
     6421 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg
     6481 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag
     6541 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact
     6601 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac
     6661 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     6721 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     6781 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     6841 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     6901 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     6961 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     7021 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     7081 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7141 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     7201 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     7261 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc
     7321 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7381 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7441 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7501 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7561 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7621 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7681 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7741 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7801 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7861 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7921 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7981 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     8041 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     8101 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     8161 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     8221 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8281 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8401 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8461 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8521 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8581 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8641 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8701 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8761 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8821 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8881 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8941 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     9001 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     9061 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9121 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     9181 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9241 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9301 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9361 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9421 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9481 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt
     9541 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     9601 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     9661 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
     9721 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
     9781 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
