LOCUS       sequence-412369-       12387 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Derived from LentiCRISPRV2 by replacing Cas9 with Cre recombinase 
            and puromycin resistance with GpNLuc. Contains a stuffer sequence 
            for cloning of sgRNA of interest, as per LentiCRISPRV2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12387)
  AUTHORS   Cho MG, Kumar RJ, Lin CC, Boyer JA, Shahir JA, Fagan-Solis K, 
            Simpson DA, Fan C, Foster CE, Goddard AM, Lerner LM, Ellington SW, 
            Wang Q, Wang Y, Ho AY, Liu P, Perou CM, Zhang Q, McGinty RK, Purvis 
            JE, Gupta GP
  TITLE     MRE11 liberates cGAS from nucleosome sequestration during 
            tumorigenesis.
  JOURNAL   Nature. 2024 Jan 10. doi: 10.1038/s41586-023-06889-6.
  PUBMED    38200309
REFERENCE   2  (bases 1 to 12387)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2024 Jan 10. doi: 10.1038/s41586-023-06889-6."
COMMENT     Sequence Label: Lenti-sgRNA-Cre-GpNLuc
FEATURES             Location/Qualifiers
     source          1..12387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      932..1520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1421..1440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1674..1691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1808..1829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1844..1849,1850..1867,1868..1874)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1882..1898
                     /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     1887..1909
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1906..1922
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1906..1922
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1971..2092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1982..2001)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2036..2055
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2222..2596)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(2615..2662)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2710..3039)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2724..2859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2778..2797)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3024..3044
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3053..3481)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3163..3184)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3375..3394
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3527..3751
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3740..3757
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             3783..4016
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    4088..4676
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4194..4205
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4603..4623
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4692..5207)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             complement(join(5223..5933,5934..5936,5937..5939))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5223..5933,5934..5936,5937..5939))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5265..5286)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5614..5633
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(5940..5996)
                     /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             complement(5997..7022)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     CDS             complement(7023..7043)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(7069..7280)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(7342..7417)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(9299..9547)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(9357..9376)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(9527..9547)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    9598..9715
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(10013..10057)
                     /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    10242..10475
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    10968..11093
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11140..11320
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(11338..11536)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(11363..11383)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        11538..11917
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     12089..12108
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        12183..12287
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12288..12356,12357..12387,1..761)
                     /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"
ORIGIN
        1 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
       61 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      121 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      181 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      241 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      301 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      361 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      421 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      481 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      541 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      601 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      661 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
      721 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
      781 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      841 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      901 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      961 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1021 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1081 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1141 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1201 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1261 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1321 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1381 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1441 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1501 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1561 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1621 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1681 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     1741 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1801 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1861 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1921 accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat
     1981 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2041 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg
     2101 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     2161 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtagcacgt
     2221 gtcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga
     2281 actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga
     2341 agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca
     2401 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca
     2461 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc
     2521 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac
     2581 tggtcaactt ggccatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact
     2641 atgccgatga ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc
     2701 cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     2761 ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     2821 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     2881 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     2941 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     3001 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg
     3061 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3121 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3181 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3241 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3301 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3361 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3421 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3481 tggggatacc ccctagagcc ccagctggtt ctttccgcct cagaagccat agagcccacc
     3541 gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     3601 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     3661 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     3721 acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgggt ttaaacgggc
     3781 cctgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     3841 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     3901 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     3961 cccagtacaa gcaaaaagca gatcttgtct tcgttgggag tgaattagcc cttccagtcc
     4021 ccccttttct tttaaaaagt ggctaagatc tacagctgcc ttgtaagtca ttggtcttaa
     4081 agtcgacgcg gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga
     4141 cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg
     4201 gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc
     4261 agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc
     4321 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg
     4381 agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg
     4441 ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac
     4501 tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     4561 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     4621 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattgtcg
     4681 acttaacgcg tttacgccag aatgcgttcg cacagccgcc agccggtcac tccgttgatg
     4741 gttactcgga acagcaggga gccgtcgggg ttgatcaggc gctcgtcgat aattttgttg
     4801 ccgttccaca gggtccctgt tacagtgatc tttttgccgt cgaacacggc gatgccttca
     4861 tacggccgtc cgaaatagtc gatcatgttc ggcgtaaccc cgtcgattac cagtgtgcca
     4921 tagtgcagga tcaccttaaa gtgatgatca tccacagggt acaccacctt aaaaattttt
     4981 tcgatctggc ccatttggtc gccgctcaga ccttcatacg ggatgatgac atggatgtcg
     5041 atcttcagcc cattttcacc gctcaggaca atcctttgga tcggagttac ggacaccccg
     5101 agattctgaa acaaactgga cacacctccc tgttcaagga cttggtccag gttgtagccg
     5161 gctgtctgtc gccagtcccc aacgaaatct tcgagtgtga agaccatacc tccggagata
     5221 tccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     5281 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     5341 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     5401 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     5461 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     5521 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     5581 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     5641 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     5701 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     5761 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     5821 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     5881 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatc
     5941 ggtccaggat tctcttcgac atctccggct tgtttcagca gagagaagtt tgttgcatcg
     6001 ccatcttcca gcaggcgcac cattgcccct gtttcactat ccaggttacg gatatagttc
     6061 atgacaatat ttacattggt ccagccacca gcttgcatga tctccggtat tgaaactcca
     6121 gcgcgggcca tatctcgcgc ggctccgaca cgggcactgt gtccagacca ggccaggtat
     6181 ctctgaccag agtcatcctt agcgccgtaa atcaatcgat gagttgcttc aaaaatccct
     6241 tccagggcgc gagttgatag ctggctggtg gcagatggcg cggcaacacc attttttctg
     6301 acccggcaaa acaggtagtt attcggatca tcagctacac cagatacgga aatccatcgc
     6361 tcgaccagtt tagttacccc caggctaagt gccttctcta cacctgcggt gctaaccagc
     6421 gttttcgttc tgccaatatg gattaacatt ctcccaccgt cagtacgtga gatatcttta
     6481 accctgatcc tggcaatttc ggctatacgt aacagggtgt tataagcaat ccccagaaat
     6541 gccagattac gtatatcctg gcagcgatcg ctattttcca tgagtgaacg aacctggtcg
     6601 aaatcagtgc gttcgaacgc tagagcctgt tttgcacgtt caccggcatc aacgttttct
     6661 tttcggatcc gccgcataac cagtgaaaca gcattgctgt cacttggtcg tggcagcccg
     6721 gaccgacgat gaagcatgtt tagctggccc aaatgttgct ggatagtttt tactgccaga
     6781 ccgcgcgcct gaagatatag aagataatcg cgaacatctt caggttctgc gggaaaccat
     6841 ttccggttat tcaacttgca ccatgccgcc cacgaccggc aaacggactg aagcattttc
     6901 caggtatgct cagaaaacgc ctggcgatcc ctgaacatgt ccatcaggtt cttgcgaacc
     6961 tcatcactcg ttgcatcgac cggtaatgca ggcaaatttt ggtgtacggt cagtaaattg
     7021 gacaccttcc tcttcttctt gggcatggta gatcctctag aaccggtcct gtgttctggc
     7081 ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat acggttctcc
     7141 cccaccctcg ggaaaaaggc ggagccagta cacgacatca ctttcccagt ttaccccgcg
     7201 ccaccttctc taggcaccgg atcaattgcc gacccctccc cccaacttct cggggactgt
     7261 gggcgatgtg cgctctgccc actgacgggc accggagcca attcccactc ctttcaagac
     7321 ctagctagcg aattcaaaaa agcaccgact cggtgccact ttttcaagtt gataacggac
     7381 tagccttatt ttaacttgct atttctagct ctaaaacaga gacgtacaaa aaagagcaag
     7441 aagctaaaaa agatttaaaa attattttta gcgcagttaa tggaacagga actaaattta
     7501 ccccaaaaat attacgtgaa tcaggatata acgttattga ggttgaagag catgcatttg
     7561 aagatgaaac atttaaaaat gttgtaaatc caaatccaga atttgatcct gcatgaaaaa
     7621 taccgcttga atatggtatt aaacatgatg cagatattat tattatgaat gacccagatg
     7681 ctgacagatt tggaatggca ataaaacatg atggtcattt tgtaagatta gatggaaatc
     7741 aaacaggacc aattttaatt gattgaaaat tatcaaatct aaaacgctta aatagcattc
     7801 caaaaaatcc ggctctatat tcaagttttg taacaagtga tttgggtgat agaatcgctc
     7861 atgaaaaata tggagttaat attgtaaaaa ctttaactgg atttaaatga atgggtagag
     7921 aaattgctaa agaagaagat aacggattaa attttgtttt tgcttatgaa gaaagttatg
     7981 gatatgtaat tgatgactca gctagagata aagatggaat acaagcttct atattaatag
     8041 cagaggctgc ttgattttat aaaaaacaaa ataaaacatt agtagactat ttagaagatt
     8101 tatttaaaga aatgggtgca tattacactt tcactttaaa cttgaatttt aaaccagaag
     8161 aaaagaaatt aaaaattgaa ccattaatga aatcattgag agcaacaccc ttaactcaaa
     8221 ttgctggact taaagttgtt aatgttgaag actacatcga tggaatgtat aatatgccag
     8281 gacaagactt actaaaattt tatttagaag ataagtcatg atttgctgtt cgcccaagtg
     8341 gaactgaacc taaactaaaa atttatttta taggtgttgg tgaatctgtt caaaacgcta
     8401 aagttaaagt agacgaaatt attaaagaat taaaattaaa aatgaatata taggagaaaa
     8461 aatgaaacta aacaaatata tagatcacac attattaaaa caagatgcta cgaaagctga
     8521 aattaaacaa ttatgtgatg aagcaattga atttgatttt gcaacagttt gtgttaattc
     8581 atattgaaca agctattgta aagaattatt aaaaggcaca aatgtaggaa taacaaatgt
     8641 tgtaggtttt cctctaggtg catgcacaac agctacaaaa gcattcgaag tttctgaagc
     8701 aattaaagat ggtgcaacag aaattgatat ggtattaaat attggtgcat taaaagacaa
     8761 aaattatgaa ttagttttag aagacatgaa agctgtaaaa aaagcagctg gatcacatgt
     8821 tgttaaatgt attatggaaa attgtttatt aacaaaagaa gaaatcatga aagcttgtga
     8881 aatagctgtt gaagctggat tagaatttgt taaaacatca acaggatttt caaaatcagg
     8941 tgcaacattt gaagatgtta aactaatgaa gtcagttgtt aaagacaatg ctttagttaa
     9001 agcagctggt ggagttagaa catttgaaga tgctcaaaaa atgattgaag caggagctga
     9061 ccgcttagga acaagtggtg gagtagctat tattaaaggt gaagaaaaca acgcgagtta
     9121 ctaaaactag cgttttttta ttttgctcat ttttattaaa agtttgcaaa aaggaacata
     9181 aaaattctaa ttattgatac taaagttatt aaaaagaaga ttttggttga ttttataaag
     9241 gtcatagaat ataatatttt agcatgtgta ttttgtgtgc tcatttacaa ccgtctccgg
     9301 tgtttcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg
     9361 taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt
     9421 attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc
     9481 taattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata
     9541 ggccctcggt accttaatta accaaactgg atctctgctg tccctgtaat aaacccgaaa
     9601 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat
     9661 gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg
     9721 gatgaatact gccatttgtc tgcagaattg gcgcacgcag tgccgatccg ttcactaatc
     9781 gaatggatct gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg
     9841 gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc
     9901 tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca
     9961 ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg
    10021 cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt
    10081 caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg
    10141 tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag
    10201 cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta
    10261 ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt
    10321 gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac
    10381 cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc
    10441 ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct
    10501 gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt
    10561 tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga
    10621 tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct
    10681 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact
    10741 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc
    10801 agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct
    10861 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt
    10921 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac
    10981 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc
    11041 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct
    11101 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg
    11161 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
    11221 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
    11281 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca aaacgcgctg
    11341 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
    11401 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
    11461 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
    11521 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
    11581 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
    11641 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
    11701 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
    11761 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
    11821 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
    11881 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca
    11941 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
    12001 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
    12061 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
    12121 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
    12181 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
    12241 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
    12301 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
    12361 cagaaacgct ggtgaaagta aaagatg
//
