LOCUS       pLenti_hU6-peg(ATP1A1-T 9884 bp ds-DNA     circular     04-FEB-2025
DEFINITION  .
KEYWORDS    "accession:addgene_84752_164464"
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported from database: AddGene Entry: Lenti_gRNA-Puro URL:
            https://www.addgene.org/browse/sequence/164464/ GENERAL Depositing
            Lab: Hyongbum Kim Publication: Kim et al Nat Methods. 2016 Dec 19.
            doi: 10.1038/nmeth.4104. Purpose: (Empty Backbone) Expresses Cpf1
            customizable sgRNA from U6 promoter and puromycin resistance from
            EF-1a promoter BACKBONE Backbone size (bp): 8330 Promoter: hU6
            Selectable markers: Puromycin Vector backbone:
            LentiGuide-Puro(Addgene#:52963) Vector type: Mammalian Expression,
            Lentiviral, CRISPR CLONING INFORMATION 3′ sequencing primer:
            hGata4-rev (5'-ATTGTGGATGAATACTGCC-3') 5′ sequencing primer: hU6-F
            Cloning method: Restriction Enzyme GROWTH IN BACTERIA Bacterial
            Resistance(s): Ampicillin Copy number: High Copy Growth Strain(s):
            NEB Stable Growth Temperature: 37°C Growth instructions: Use SapI
            digest to check for unwanted recombination of lentiviral plasmid.
            Only amplify in RecA- bacteria (eg. Stbl3). TERMS AND LICENSES
            Academic/Nonprofit Terms:  Industry Terms:
FEATURES             Location/Qualifiers
     primer_bind     115..132
                     /label="L4440"
                     /note="note: L4440 vector, forward primer"
     protein_bind    249..270
                     /label="CAP binding site"
                     /note="bound_moiety: E. coli catabolite activator protein note: CAP binding activates transcription in the presence of cAMP."
     promoter        285..315
                     /label="lac promoter"
                     /note="note: promoter for the E. coli lac operon"
     protein_bind    323..339
                     /label="lac operator"
                     /note="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     328..350
                     /label="M13/pUC Reverse"
                     /note="note: In lacZ gene"
     primer_bind     347..363
                     /label="M13 rev"
                     /note="note: common sequencing primer, one of multiple similar variants"
     primer_bind     347..363
                     /label="M13 Reverse"
                     /note="note: In lacZ gene. Also called M13-rev"
     primer_bind     382..402
                     /label="T3"
                     /note="note: T3 promoter, forward primer"
     promoter        384..402
                     /label="T3 promoter"
                     /note="note: promoter for bacteriophage T3 RNA polymerase"
     promoter        430..656
                     /label="RSV promoter"
                     /note="note: Rous sarcoma virus enhancer/promoter"
     LTR             657..837
                     /label="5' LTR (truncated)"
                     /note="note: truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    884..1009
                     /label="HIV-1 Psi"
                     /note="note: packaging signal of human immunodeficiency virus type 1"
     misc_feature    1502..1735
                     /label="RRE"
                     /note="note: The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm."
     CDS             1920..1964
                     /label="gp41 peptide"
                     /note="product: antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013) note: recognized by the 2H10 single-chain llama nanobody translation: KNEQELLELDKWASL"
     CDS             1920..1964
                     /translation="KNEQELLELDKWASL"
     primer_bind     2142..2162
                     /label="hU6-F"
                     /note="note: Human U6 promoter, forward primer"
     promoter        2142..2390
                     /label="U6 promoter"
                     /note="note: RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2313..2332
                     /label="LKO.1 5'"
                     /note="note: Human U6 promoter, forward primer"
     misc_feature    2391..2410
                     /label="spacer"
     misc_feature    2391..2569
                     /label="pegRNA(ATP1A1-T804N)"
     misc_feature    2411..2496
                     /label="scaffold"
     misc_feature    2497..2512
                     /label="RTT"
     misc_feature    2513..2525
                     /label="PBS"
     misc_feature    2526..2562
                     /label="tevopreQ1"
     misc_feature    2563..2569
                     /label="polyT"
     misc_feature    2654..2974
                     /label="mU6 promoter"
     misc_feature    2975..4002
                     /label="BsmBI cassette"
     gene            3040..3828
                     /label="NeoR/KanR"
     misc_feature    4003..4027
                     /label="Adapter"
     misc_feature    4104..4221
                     /label="cPPT/CTS"
                     /note="note: central polypurine tract and central termination sequence of HIV-1"
     promoter        4360..5538
                     /label="EF-1-alpha promoter"
                     /note="note: strong constitutive promoter for human elongation factor EF-1-alpha"
     intron          4591..5529
                     /label="EF-1-alpha intron A"
                     /note="note: intron upstream of the start codon of human EF-1-alpha"
     primer_bind     5486..5506
                     /label="EF1a-F"
                     /note="note: Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(5551..5570)
                     /label="Puro-R"
                     /note="note: Puromycin resistance gene, reverse primer. Also called puro-variant-R"
     CDS             5551..6150
                     /label="PuroR"
                     /note="gene: pac from Streptomyces alboniger product: puromycin N-acetyltransferase note: confers resistance to puromycin translation: MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             5551..6150
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
     primer_bind     6047..6067
                     /label="Puro-F"
                     /note="note: Puromycin resistance gene, forward primer"
     misc_feature    6165..6754
                     /label="WPRE"
                     /note="note: woodchuck hepatitis virus posttranscriptional regulatory element"
     primer_bind     complement(6219..6239)
                     /label="WPRE-R"
                     /note="note: WPRE, reverse primer"
     CDS             complement(6637..6648)
                     /label="Factor Xa site"
                     /note="product: Factor Xa recognition and cleavage site translation: IEGR"
     CDS             complement(6637..6648)
                     /translation="IEGR"
     LTR             6826..7059
                     /label="3' LTR (Delta-U3)"
                     /note="note: self-inactivating 3' long terminal repeat (LTR) fromHIV-1"
     polyA_signal    7137..7258
                     /label="SV40 poly(A) signal"
                     /note="note: SV40 polyadenylation signal"
     primer_bind     complement(7174..7193)
                     /label="SV40pA-R"
                     /note="note: SV40 polyA, reverse primer"
     primer_bind     7228..7247
                     /label="EBV-rev"
                     /note="note: SV40 polyA terminator, reverse primer"
     rep_origin      7298..7433
                     /label="SV40 ori"
                     /note="note: SV40 origin of replication"
     primer_bind     7360..7379
                     /label="SV40pro-F"
                     /note="note: SV40 promoter/origin, forward primer"
     primer_bind     complement(7453..7472)
                     /label="T7"
                     /note="note: T7 promoter, forward primer"
     promoter        complement(7454..7472)
                     /label="T7 promoter"
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7482..7498)
                     /label="M13 fwd"
                     /note="note: common sequencing primer, one of multiple similar variants"
     primer_bind     complement(7482..7499)
                     /label="M13 Forward"
                     /note="note: In lacZ gene. Also called M13-F20 or M13 (-21) Forward"
     primer_bind     complement(7491..7513)
                     /label="M13/pUC Forward"
                     /note="note: In lacZ gene"
     rep_origin      7640..8095
                     /label="f1 ori"
                     /note="direction: RIGHT note: f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
     primer_bind     complement(7727..7746)
                     /label="F1ori-R"
                     /note="note: F1 origin, reverse primer"
     primer_bind     7937..7958
                     /label="F1ori-F"
                     /note="note: F1 origin, forward primer"
     promoter        8121..8225
                     /label="AmpR promoter"
                     /note="gene: bla"
     CDS             8226..9086
                     /label="AmpR"
                     /note="gene: bla product: beta-lactamase note: confers resistance to ampicillin, carbenicillin, andrelated antibiotics translation: MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     CDS             8226..9086
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer_bind     complement(8444..8463)
                     /label="Amp-R"
                     /note="note: Ampicillin resistance gene, reverse primer"
     rep_origin      9257..9845
                     /label="ori"
                     /note="direction: RIGHT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     primer_bind     9746..9765
                     /label="pBR322ori-F"
                     /note="note: pBR322 origin, forward primer"
ORIGIN
        1 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
       61 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
      121 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
      181 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
      241 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
      301 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
      361 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
      421 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
      481 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
      541 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
      601 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
      661 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
      721 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
      781 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
      841 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
      901 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
      961 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     1021 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     1081 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     1141 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     1201 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     1261 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     1321 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     1381 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1441 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1501 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1561 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1621 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1681 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1741 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     1801 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     1861 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     1921 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     1981 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     2041 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     2101 tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta tttcccatga
     2161 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
     2221 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     2281 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     2341 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc GCCCAAGTCA
     2401 ATGCAGAGGA GTTTAAGAGC TATGCTGGAA ACAGCATAGC AAGTTTAAAT AAGGCTAGTC
     2461 CGTTATCAAC TTGAAAAAGT GGCACCGAGT CGGTGCGGGA aTGTCACtAT CCTCTGCATT
     2521 GACTTCGCGG TTCTATCTAG TTACGCGTTA AACCAACTAG AAtttttttc tactctatct
     2581 agacgagacc gtctactcta ccacttgtac ttcagcggtc agcttactcg acttaacgtg
     2641 cacgtgacac gtttctagag atccgacgcc gccatctcta ggcccgcgcc ggccccctcg
     2701 cacagacttg tgggagaagc tcggctactc ccctgccccg gttaatttgc atataatatt
     2761 tcctagtaac tatagaggct taatgtgcga taaaagacag ataatctgtt ctttttaata
     2821 ctagctacat tttacatgat aggcttggat ttctataaga gatacaaata ctaaattatt
     2881 attttaaaaa acagcacaaa aggaaactca ccctaactgt aaagtaattg tgtgttttga
     2941 gactataaat atgcatgcga gaaaagcctt gtttggagac gggaggccta ggcttttgca
     3001 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3061 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3121 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3181 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3241 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3301 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3361 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3421 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3481 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3541 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     3601 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3661 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3721 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3781 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3841 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3901 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3961 tcctccagcg cggggatctc atgctggagt tcttccgtct cgggtaagct cgtaccgtga
     4021 ttcatgccta gaccgtacat gcttacatgg gatgaagctt ggcgtaacta gatcttgaga
     4081 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     4141 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     4201 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggcgc
     4261 cggctcgagg gggcccgggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     4321 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     4381 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc
     4441 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     4501 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4561 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
     4621 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
     4681 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     4741 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     4801 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     4861 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     4921 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     4981 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     5041 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     5101 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     5161 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     5221 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     5281 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     5341 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     5401 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     5461 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     5521 ccatttcagg tgtcgtgacg tacggccacc atgaccgagt acaagcccac ggtgcgcctc
     5581 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5641 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5701 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5761 gccgccgtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5821 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5881 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc
     5941 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     6001 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     6061 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     6121 tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat
     6181 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     6241 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     6301 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     6361 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     6421 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     6481 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     6541 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     6601 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     6661 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6721 acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca
     6781 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     6841 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     6901 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6961 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7021 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc
     7081 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     7141 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7201 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7261 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7321 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7381 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac
     7441 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     7501 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     7561 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     7621 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7681 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     7741 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7801 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     7861 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7921 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     7981 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8041 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     8101 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     8161 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     8221 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     8281 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     8341 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     8401 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     8461 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     8521 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     8581 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     8641 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     8701 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     8761 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     8821 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     8881 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8941 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     9001 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     9061 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     9121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     9181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     9241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     9301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     9361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     9421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     9481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     9541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     9601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     9661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     9721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     9781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     9841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttt
//