LOCUS       sequence-119608-        8112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR gRNA expression vector with the conventional scaffold and 
            puro/BFP markers without WPRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8112)
  AUTHORS   Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, 
            Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi 
            S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli 
            P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly 
            BJ, Iorio F, Pina C, Vassiliou GS, Yusa K
  TITLE     A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and 
            Therapeutic Targets in Acute Myeloid Leukemia.
  JOURNAL   Cell Rep. 2016 Oct 18;17(4):1193-1205. doi: 
            10.1016/j.celrep.2016.09.079.
  PUBMED    27760321
REFERENCE   2  (bases 1 to 8112)
  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.2016.09"; journalName: "Cell Rep"; date: 
            "2016-10-18- 18"; volume: "17"; issue: "4"; pages: "1193-1205"
COMMENT     Sequence Label: pKLV2-U6gRNA3(BbsI)-PGKpuro2ABFP
FEATURES             Location/Qualifiers
     source          1..8112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        715..1018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1020..1218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1173..1193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1236..1416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1463..1588
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2087..2320
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2505..2549
                     /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    2847..2964
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3039..3287
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3039..3059
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3210..3229
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3306..3381
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3404..3903
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3439..3462)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3836..3855
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3943..4539
                     /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(3943..3962)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4439..4459
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4558..4611
                     /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             4612..5313
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(5668..5685)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5674..5898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5924..5944)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5924..5942)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5963..5979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5963..5979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5976..5998)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5987..6003
                     /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(6011..6017,6018..6035,6036..6041))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6056..6077
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6194..6211)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6365..6953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6445..6464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7124..7915,7916..7984))
                     /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     7747..7766
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7985..8089)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgactagtta
      661 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      721 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      781 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      841 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      901 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      961 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1021 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1081 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1141 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1201 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     1261 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1321 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1381 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     1441 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     1501 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1561 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1621 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1681 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1741 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1801 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1861 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1921 gcaagcggcc ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1981 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     2041 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2101 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2161 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     2221 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     2281 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2341 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2401 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2461 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2521 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2581 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2641 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2701 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2761 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2821 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2881 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2941 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     3001 ttagtaccgg gccctacgcg tccaaggtcg ggcaggaaga gggcctattt cccatgattc
     3061 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
     3121 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     3181 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     3241 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggg tcttcgagaa
     3301 gacacgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     3361 aaagtggcac cgagtcggtg ctttttttgg atccaattct accgggtagg ggaggcgctt
     3421 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     3481 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     3541 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     3601 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     3661 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     3721 cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg
     3781 ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     3841 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     3901 tcgacctgca tccatctaga tctcgagcag ctgaagctta ccatgaccga gtacaagccc
     3961 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     4021 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     4081 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4141 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4201 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4261 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4321 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4381 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4441 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4501 ggaccgcgca cctggtgcat gacccgcaag cccggtgccg gcggcgggtc cggaggagag
     4561 ggcagaggaa gtctcctaac atgcggtgac gtggaggaga atcctggccc aatgagcgag
     4621 ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgga caaccatcac
     4681 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc
     4741 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac tagcttcctc
     4801 tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt caagcagtcc
     4861 ttccctgagg gcttcacatg ggagagagtc accacatacg aggacggggg cgtgctgacc
     4921 gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg
     4981 gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccttc
     5041 accgagacgc tgtaccccgc tgacggcggc ctggaaggca gaaacgacat ggccctgaag
     5101 ctcgtgggcg ggagccatct gatcgcaaac atcaagacca catatagatc caagaaaccc
     5161 gctaagaacc tcaagatgcc tggcgtctac tatgtggact acagactgga aagaatcaag
     5221 gaggccaaca acgagaccta cgtcgagcag cacgaggtgg cagtggccag atactgcgac
     5281 ctccctagca aactggggca caagcttaat tgagcggccg ctaggtacct ttaagaccaa
     5341 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     5401 ggctaattca ctcccaaaga agtcaagatc tgctttttgc ctgtactggg tctctctggt
     5461 tagaccagag tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5521 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5581 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5641 ctctagcagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     5701 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     5761 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     5821 gtggggtggg gcaggacagc aagggggagg attgggaagt caatagcagg catgctgggg
     5881 atgcggtggg ctctatgggc ggccgttaat taagcttttg ttccctttag tgagggttaa
     5941 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6001 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6061 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6121 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6181 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6241 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6301 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6361 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6481 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6541 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6601 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6661 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6721 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6781 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6841 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6901 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6961 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7021 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7081 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7141 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7201 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7261 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7321 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7381 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7441 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7501 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7561 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7621 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7681 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7741 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7801 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7861 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7921 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7981 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8041 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8101 gaaaagtgcc ac
//
