LOCUS       sequence-343516-       10105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral delivery of sgRNA. Expresses PuroR and eGFP from SFFV 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10105)
  AUTHORS   Muller I, Moroni AS, Shlyueva D, Sahadevan S, Schoof EM, 
            Radzisheuskaya A, Hojfeldt JW, Tatar T, Koche RP, Huang C, Helin K
  TITLE     MPP8 is essential for sustaining self-renewal of ground-state 
            pluripotent stem cells.
  JOURNAL   Nat Commun. 2021 May 24;12(1):3034. doi: 10.1038/s41467-021-23308-4.
  PUBMED    34031396
REFERENCE   2  (bases 1 to 10105)
  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"; journalName: "Nat 
            Commun."; date: "2021-05-24"; volume: "12(1):3034. doi"; pages: " 
            10.1038/s41467-021-23308-4"
COMMENT     Sequence Label: pL-U6-sgRNA-SFFV-Puro-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..10105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             152..332
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    379..504
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    997..1230
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1415..1459
                     /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"
     promoter        1637..1885
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1637..1657
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1808..1827
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3767..3842
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3890..4007
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4065..4472
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             4478..5074
                     /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(4478..4497)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4974..4994
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5090..5146
                     /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             join(5147..5149,5150..5152,5153..5863)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5192..5213)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5453..5472)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5800..5821
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5882..6470
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5935..5955)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6353..6364)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6542..6775
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6853..6974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6890..6909)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6944..6963
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7014..7149
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7169..7188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7170..7188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7198..7215)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7198..7214)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7207..7229)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7356..7811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7443..7462)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7653..7674
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7837..7941
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7942..8010,8011..8802)
                     /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     complement(8160..8179)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8973..9561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9462..9481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9715..9732
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9849..9870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9885..9890,9891..9908,9909..9915)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9923..9939
                     /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)."
     primer_bind     9928..9950
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9947..9963
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9947..9963
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9982..10002
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9984..10002
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(10030..10105,1..151)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
       61 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
      121 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
      181 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
      241 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
      301 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
      361 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
      421 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
      481 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
      541 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
      601 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
      661 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
      721 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
      781 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
      841 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
      901 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
      961 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1021 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1081 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1141 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1201 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1261 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1321 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     1381 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     1441 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     1501 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     1561 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     1621 cccgagggga cccagagagg gcctatttcc catgattcct tcatatttgc atatacgata
     1681 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
     1741 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     1801 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     1861 tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa tgagcacaca aaatacacat
     1921 gctaaaatat tatattctat gacctttata aaatcaacca aaatcttctt tttaataact
     1981 ttagtatcaa taattagaat ttttatgttc ctttttgcaa acttttaata aaaatgagca
     2041 aaataaaaaa acgctagttt tagtaactcg cgttgttttc ttcaccttta ataatagcta
     2101 ctccaccact tgttcctaag cggtcagctc ctgcttcaat cattttttga gcatcttcaa
     2161 atgttctaac tccaccagct gctttaacta aagcattgtc tttaacaact gacttcatta
     2221 gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt ttaacaaatt
     2281 ctaatccagc ttcaacagct atttcacaag ctttcatgat ttcttctttt gttaataaac
     2341 aattttccat aatacattta acaacatgtg atccagctgc tttttttaca gctttcatgt
     2401 cttctaaaac taattcataa tttttgtctt ttaatgcacc aatatttaat accatatcaa
     2461 tttctgttgc accatcttta attgcttcag aaacttcgaa tgcttttgta gctgttgtgc
     2521 atgcacctag aggaaaacct acaacatttg ttattcctac atttgtgcct tttaataatt
     2581 ctttacaata gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca aattcaattg
     2641 cttcatcaca taattgttta atttcagctt tcgtagcatc ttgttttaat aatgtgtgat
     2701 ctatatattt gtttagtttc attttttctc ctatatattc atttttaatt ttaattcttt
     2761 aataatttcg tctactttaa ctttagcgtt ttgaacagat tcaccaacac ctataaaata
     2821 aatttttagt ttaggttcag ttccacttgg gcgaacagca aatcatgact tatcttctaa
     2881 ataaaatttt agtaagtctt gtcctggcat attatacatt ccatcgatgt agtcttcaac
     2941 attaacaact ttaagtccag caatttgagt taagggtgtt gctctcaatg atttcattaa
     3001 tggttcaatt tttaatttct tttcttctgg tttaaaattc aagtttaaag tgaaagtgta
     3061 atatgcaccc atttctttaa ataaatcttc taaatagtct actaatgttt tattttgttt
     3121 tttataaaat caagcagcct ctgctattaa tatagaagct tgtattccat ctttatctct
     3181 agctgagtca tcaattacat atccataact ttcttcataa gcaaaaacaa aatttaatcc
     3241 gttatcttct tctttagcaa tttctctacc cattcattta aatccagtta aagtttttac
     3301 aatattaact ccatattttt catgagcgat tctatcaccc aaatcacttg ttacaaaact
     3361 tgaatataga gccggatttt ttggaatgct atttaagcgt tttagatttg ataattttca
     3421 atcaattaaa attggtcctg tttgatttcc atctaatctt acaaaatgac catcatgttt
     3481 tattgccatt ccaaatctgt cagcatctgg gtcattcata ataataatat ctgcatcatg
     3541 tttaatacca tattcaagcg gtatttttca tgcaggatca aattctggat ttggatttac
     3601 aacattttta aatgtttcat cttcaaatgc atgctcttca acctcaataa cgttatatcc
     3661 tgattcacgt aatatttttg gggtaaattt agttcctgtt ccattaactg cgctaaaaat
     3721 aatttttaaa tcttttttag cttcttgctc ttttttgtac gtctctgttt tagagctaga
     3781 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     3841 gcttttttga attctagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa
     3901 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     3961 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     4021 gggacagcag agatccactt tggcgccggc tcgagcgagc tgcagtaacg ccattttgca
     4081 aggcatggaa aaataccaaa ccaagaatag agaagttcag atcaagggcg ggtacatgaa
     4141 aatagctaac gttgggccaa acaggatatc tgcggtgagc agtttcggcc ccggcccggg
     4201 gccaagaaca gatggtcacc gcagtttcgg ccccggcccg aggccaagaa cagatggtcc
     4261 ccagatatgg cccaaccctc agcagtttct taagacccat cagatgtttc caggctcccc
     4321 caaggacctg aaatgaccct gcgccttatt tgaattaacc aatcagcctg cttctcgctt
     4381 ctgttcgcgc gcttctgctt cccgagctct ataaaagagc tcacaacccc tcactcggcg
     4441 cgccagtcct ccgacagact gagtcggatc cgccaccatg accgagtaca agcccacggt
     4501 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4561 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4621 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4681 cgacggcgcc gccgtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4741 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4801 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4861 cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4921 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     4981 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5041 gcgcacctgg tgcatgaccc gcaagcccgg tgccgctagc ggcagcggcg ccaccaactt
     5101 cagcctgctg aagcaggccg gcgacgtgga ggagaacccc ggccccatgg tgtctaaggg
     5161 cgaagagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     5221 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     5281 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     5341 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     5401 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     5461 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     5521 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     5581 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     5641 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     5701 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     5761 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     5821 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaacgc gttaagtcga
     5881 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5941 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6001 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6061 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6121 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6181 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6241 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     6301 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6361 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6421 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     6481 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6541 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     6601 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6661 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6721 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     6781 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6841 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6901 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6961 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     7021 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     7081 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     7141 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     7201 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     7261 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     7321 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     7381 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     7441 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     7501 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     7561 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     7621 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     7681 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     7741 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     7801 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     7861 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7921 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     7981 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     8041 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     8101 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     8161 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     8221 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     8281 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     8341 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     8401 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     8461 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     8521 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     8581 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     8641 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     8701 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     8761 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     8821 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     8881 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8941 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     9001 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     9061 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     9121 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     9181 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     9241 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     9301 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     9361 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     9421 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     9481 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     9541 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     9601 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     9661 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     9721 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     9781 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     9841 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     9901 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     9961 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
    10021 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
    10081 agcaacatgc cttacaagga gagaa
//
